Objective:To investigate the effect of di(2-ethylhexyl) phthalate (DEHP) on the uterus tissue of female rats, and to clarify the mechanism of uterine toxicity of DEHP in the female rats. Methods:A total of 48 Wistar female rats were randomly divided into control group(given corn oil), low, middle, and high doses of DEHP groups (given 300, 1 000, and 3 000 mg·kg^-1·d^-1 DEHP;1/100,1/30,and 1/10 LD₅₀)(n=12). The rats in various groups were sacrificed during anoestrum, the body weights were recorded and uterine organ coefficients of the rats in various groups were calculated. The pathomorphology of the uterus tissue of the rats in various groups was detected by HE staining. The expression levels of follicle stimulating hormonereceptor (FSHR) and luteinizing hormone receptor (LHR) in uterus tissue of the rats in various groups were detected by immunohistochemistry. Results:Compared with control group, the body weights of the rats in different doses of DEHP groups were decreased (P<0.05) with a dose-effect relationship; there were no significant differences in uterine organ coefficients of the rats in various groups (P>0.05). Macroscopically, the morphology of the uterus of rats in control group was normal; in middle and high doses of DEHP groups, the uterus showed distorted and the tube wall became thinner; different degrees of hydrops, congestion and swelling were seen on the serosal surface of the uterus. The microscopic observation results showed that in different doses of DEHP groups the endometrium nuclei appeared pseudostratified phenomenon; the epithelial hyperplasia and endothelial fibrosis were found in the endometrium; the gland number in the lamina propria was decreased,and part of the glands were depauperate. Compared with control group, the expression levels of FSHR in uterus tissue of the rats in different doses of DEHP groups had no significant differences(P>0.05),and the expression levels of LHR were decreased(P<0.05); the expression levels of LHR in uterus tissue of the rats in middle and high doses of DEHP groups were significantly lower than that in low dose of DEHP group (P<0.05). Conclusion:DEHP can lead to the weight loss and the pathological changes in the uterus tissue of the female rats, and has a uterine toxicity effect on the female rats.

Objective:To establish the glioma-bearing nude mouse models, and to investigate the effect of HOXA4 on the growth of glioma U251 cells in vivo and its regulatory effect on the Wnt/β-catenin signal pathway and its mechanism. Methods:The glioma U251 cell line stably transfected with HOXA4 siRNA(si-HOXA4) and the U251 cell line stably transfected with blank vector (si-NC) were established by lentivirus transfection. The U251, si-NC, and si-HOXA4 cells were respectively inoculated under the skin of the neck and back of the BALB/c nude mice to establish the glioma-bearing nude mouse models named as control group, si-NC group, and si-HOXA4 group. The tumorigenesis of nude mice in various groups were observed and the tumor growth curve was drawn. The tumor tissue was stripped after the mice were sacrificed on the 21 th day, and the volume and weight of tumor were measured; the relative mRNA expression amounts of HOXA4, CTNNB1, and Gsk3β in tumor tissue of the nude mice in various groups were detected by qRT-PCR method; the expression levels of HOXA4, β-catenin, Gsk3β, CyclinD1, and P53 proteins in tumor tissue of the nude mice in various groups were detected by immunohistochemistry (IHC) method. Results:Compared with si-NC group and control group, the volume and weight of tumor of the nude mice in si-HOXA4 group were significantly decreased(P<0.05). The relative expression amount of HOXA4 mRNA and the expression level of HOXA4 protein in si-HOXA4 group were significantly lower than those in the other groups (P<0.05). Compared with si-NC group and control group, the relative expression amount of CTNNB1 mRNA and the expression levels of β-catenin and CyclinD1 proteins in si-HOXA4 group were significantly decreased (P<0.05), and the expression levels of Gsk3β and P53 proteins were significantly increased (P<0.05). Conclusion:Inhibition of HOXA4 expression in human glioma U251 cells can regulate the expressions of CyclinD1 and P53 through Wnt/-catenin signal pathway in vivo, thus inhibiting the tumor growth of glioma-bearing nude mice.

Objective:To investigate the protective effect of panaxdiol saponins(PDS) on the acute liver damage(ALD) of the rats induced by carbon tetrachloride(CCl₄), and to clarify its mechanism. Methods:Fifty rats were randomly divided into control group (the rats were given normal saline), model group (the ALD models were established and given normal saline) and low,middle, and high doses (10, 20,and 40 mg·kg^-1) of PDS groups, and there were 10 rats in each group.The general state of the rats in various groups was observed. The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),and total protein(TP) of the rats in various groups were detected; ELISA method was used to detecet the levels of interleukin-6 (IL-6), interleukin-12 (IL-12)and tumor necrosis factor-α (TNF-α) in liver tissue homogenate of the rats in various groups; Western blotting method was used to detecet the expression levels of protein kinase B (AKT)and p-protein kinase B(p-AKT) in liver tissue of the rats in various groups. Results:Compared with control group, the levels of ALT,AST and TBIL of the rats in model group were increased(P<0.01),and the level of TP was decreased(P<0.01); the levels of IL-12, IL-6 and TNF-α were increased (P<0.01). Compared with model group, the hepatic function of the rats in different doses of PDS groups were obviously improved, the levels of ALT, AST and TBIL in liver tissue of the rats in different doses of PDS groups were decreased(P<0.01), the levels of TP were increased(P<0.01);the levels of IL-12, IL-6 and TNF-α in liver tissue of the rats were decreased (P<0.01); the expression levels of AKT protein in liver tissue of the rats were decreased (P<0.01) and the expression level of p-AKT was increased(P<0.01). Conclusion:PDS has the protective effect on ALD,and its mechanism may be related to reducing the expressions of inflammatory factors and affecting the p-AKT-AKT signal pathway.

Objective:To discuss the antiviral effect, the inhibitory effect on LINE-1 retrotransposon activity and the redection of interferon production signal pathway of restriction factor SAMHD1 of the primates,and to provide the basis for further study of the SAMHD1 of the primates. Methods:The U937 cells stably expressing the SAMHD1 of primates were established; the U937-control cells established with pLVX-puro were used as negative control group,and the U937-SAMHD1 cells stably expressing the SAMHD1 protein of the different primates were used as experimental groups; the cells were treated with PMA to induce cell differentiation. The virus infection rates of HIV-1 in the cells in various groups were determined by flow cytometry. The HEK293T cells transfected with the expression plasmid of SAMDH1 were used as control group, and the cells co-transfected with the SAMHD1 and HIV-2/SIV Vpx expression plasmids were used as experimental groups.The cells were obtained 48 h after transfection,and the expression levels of SAMHD1 protein were determined by Western blotting method. The intracellular location of SAMHD1 protein was determined by immunofluorescence.The HEK293T cells transfected with LINE-1-GFP report plasmid were used as control group, and the cells co-transfected with LINE-1-GFP and SAMHD1 expression plasmids were used as experimental groups. The rates of GFP positive cells(activity of SAMHD1 to LINE-1 transposon) were determined by flow cytometry. The HEK293T cells transfected with IFN-Luc report plasmid were used as control group,and the cells co-transfected with IFN-Luc and pSAMHD1 expression plasmids were used as experimental groups. The expression levels of luciferase in HEK293T cells were determined by chemiluminescence instrument. Results:Compared with negative control group, the virus infection rates of HIV-1 in experimental groups with stable expression of SAMHD1 in the primates were significantly decreased (P<0.01). Compared with control group, the expression levels of SAMHD1 protein of the primates in experimental groups were decreased(P<0.05 or P<0.01).The immunofluorescence results showed that the SAMHD1 protein of the primates was localized in the nucleus. Compared with control group, the rates of GFP positive cells (activity of SAMHD1 to LINE-1 transposon) in experimental groups were significantly decreased (P<0.05 or P<0.01). Compared with control group, the expression levels of luciferase in the HEK293T cells in experimental groups were significantly decreased(P<0.05). Conclusion:The SAMHD1 protein of the different primates can resist the HIV-1 infection, inhibit the LINE-1 retrotransposon and antagonize the IFN production by natural immune system.

Objective:To investigate the regulatory effect of epigallocatechin-3-gallate (EGCG) on the immune imbalance of regulatory Tlymphocytes/helper T lymphocytes 17 (Treg/Th17) in the mice with obese asthma,and to provide the basis for the treatment of obese asthma by EGCG. Methods:A total of 40 C57BL/6J mice were randomly divided into normal control group (fed with chow diet), obese group (fed with high fat diet), obese asthma group[fed with high fat diet and sensitized and challenged with ovalbumin(ova)], EGCG intervention group (20 mg·kg^-1 EGCG was administered intraperitoneally 1 h before challenge of OVA) and dexamethasone intervention group (2 mg·kg^-1 dexamethasone was administered intraperitoneally 1 h before challenge of OVA), and there were 8 mice in each group. The edhanced pause(Penh) values of the mice in various groups were measured with non-invasive lung function instrument; the pathomorphology of lung tissue of the mice in various groups were observed by HE staining,and the inflammation scores were evaluated by semi-quantitative method for HE staining section; the levels of interleukin-10 (IL-10) and interleukin-17A(IL-17A) in bronchoalveolar lavage fluid (BALF) and adiponectin and leptin in serum of the mice in various groups were detected by ELISA method,and the percentages of Treg and Th17 in spleen tissue of the mice in various groups were detected by flow cytometery. Results:Compared with normal control group, the body weight of the mice in obese group was increased (P<0.05); it was 1.67 times the average weight of the mice in normal control group. Compared with normal control group, the Penh value and inflammation score of the mice in obese asthma group were increased (P<0.05); the leptin level in serum of the mice in obese asthma group was increased (P<0.05); the IL-17 level in BALF of the mice in obese asthma group was increased (P<0.05) and IL-10 level in BALF of the mice in obese asthma group was decreased (P<0.05); the percentage of Th17 in spleen tissue of the mice in obese asthma group was increased (P<0.05) and the percentage of Treg in spleen tissue of the mice in obese asthma group was decreased (P<0.05).Compared with obese asthma group, the Penh value and inflammation score of the mice in EGCG intervention group were decreased (P<0.05); the leptin level in serum of the mice in EGCG intervention group was decreased (P<0.05); the IL-17 level in BALF of the mice in EGCG intervention group was decreased (P<0.05) and IL-10 level in BALF of the mice in EGCG intervention group was increased (P<0.05); the percentage of Th17 in spleen tissue of the mice in EGCG intervention group was decreased (P<0.05) and the percentage of Treg in spleen tissue of the mice in EGCG intervention group was increased (P<0.05). Conclusion:Treg/Th17 immune imbalance exists in the mice with obese asthma. EGCG could inhibit the airway inflammation and hyperresponsiveness in the mice with obese asthma and improve the Treg/Th17 immune imbalance.

Objective:To construct the sponge vector which can target microRNA-186(miR-186),and to create a stable EA.hy926 cell 1ine that can knockdown miR-186. Methods:The miR-186 sponge sequence was chemically synthesized and cloned into lentiviral vector FV040. Then the FV040-miR-186-sponge recombinant plasmid together with the helper plasmids were cotransfected into the HEK293T cells by using lipofectamine 2000 to package lentivirus and the viral titer was determined. The FV040-control lentivirus (designated as the control group), and the FV040-miR-186-sponge (designated as the experiment group) were used to infect EA.hy926 cells for establishing stable cell lines.The fluorescent quantitative PCR (qPCR) method was used to detect the relative expression levels of miR-186 in the EA.hy926 cells in blank group,FV040-control group and FV040-miR-186 sponge group, respectively. Results:The cloned target sequence was identical with the designed miR-186 sponge sequence. The green fluorescence protein(GFP) was observed in the infected EA.hy926 cells. The lentivirus titre of viruses in the FV040-control group was 2×10⁸ TU·mL^-1, and which was 6×10⁸ TU·mL^-1 in FV040-miR-186 sponge group. The EA.hy926 cell line stably expressed miR-186-sponge was established successfully and the infection rate was as high as 95%. The qPCR results indicated that the relative expression level of miR-186 in the EA.hy926 cells in FV040-miR-186-sponge group was lower than those in blank control group and FV040-control group(P<0.01). Conclusion:The miR-186-sponge vector is successfully constructed, and the EA.hy926-miR-186-sponge cell line with the stably decreased expression of miR-186 is established successfully.

Objective:To synthesize the zinc gluconate carbon dots (Zn-CDs) by one-step hydrothermal method, and to investigate their effects on the cell imaging and inducing osteoblastic differentiation of preosteoblasts in the mice. Methods:The Zn-CDs were synthesized by one-step hydrothermal method, and the characteristics were observed and detected by transmission electron microscope (TEM), Fourier transform-infrared spectrum(FT-IR) and fluorescence spectrometer. The MC3T3-E1 cells were divided into blank control group and experimental groups; different concentrations (0.01,0.10,1.00,10.00,100.00,1 000 mg·L^-1) of Zn-CDs were added into the cells in experimental groups, and nothing was added into the cells in blank control group. MTT assay was used to determin the relative growth rate(RGR) of MC3T3-E1 cells in various groups; the imaging characteristics of MC3T3-E1 cells were observed under confocal microscope; the relative expression levels of Runt-relateed transcription factor-2(Runx2), alkaline phosphatase (ALP) and osteocalcin (OC) mRNA in the MC3T3-E1 cells in various groups were detected by qRT-PCR;the number of calcified nodules was detected by alizarin red staining. Results:The TEM results showed that the particle size of Zn-CDs was about 5.25 nm. The fluorescence spectrometer results showed that the Zn-CDs had the fluorescence properties of 360 nm ultraviolet excited light and 450 nm blue emission light,and the Zn-CDs showed the characteristics of excitation wavelength dependence.The FT-IR results showed that the surface of Zn-CDs was mainly composed of carboxyl groups and hydroxyl groups. Compared with blank control group, the RGR of MC3T3-E1 cells in 1 000.00 mg·L^-1 Zn-CDs group was decreased significantly at 24 h after co-culture(P<0.01).The results of fluorescence imaging showed that the blue, green and red fluorescence in the MC3T3-E1 cells,the outline was clear,and the fluorescence intensity of the cytoplasm was stronger than that of the nucleus after co-cultured with Zn-CDs. The qRT-PCR results showed that the relative expression levels of Runx2, ALP and OC mRNA were significantly increased with the increasing of Zn-CDs concentration. The alizarin red staining results showed that the number of calcium deposits in the MC3T3-E1 cells in different concentrations of Zn-CDs groups were more than that in blank control group after induced for 21 d. Conclusion:Zn-CDs can effectively perform the fluorescence imaging in the MC3T3-E1 cells, and Zn-CDs have a certain ability to promote the osteoblastic differentiation of the MC3T3-E1 cells.

Objective:To investigate the changes of the serum indexes of renin-angiotensin-aldosterone system(RAAS) and atrial fibrillation burden in the atrial fibrillation model dogs underwent spinal cord stimulation(SCS) therapy, and to illuminate the therapeutic effect of SCS on the atrial fibrillation and its effect on RAAS. Methods:A total of 24 healthy adult dogs were chosen.The pacemakers and miniaturized insertable cardiac monitors were implanted into 16 dogs to establish the atrial fibrillation models by rapid right atrial pacing. Then 16 model dogs were divided into atrial fibrillation (AF) group and SCS group(n=8). The spinal cord stimulators were implanted into the dogs in SCS group to release stimulation for 12 weeks. And the rest dogs were regareded as blank control group(n=8). The atrial fibrillation burden of dogs in AF group and SCS group was monitored by miniaturized insertable cardiac monitors;the left and right atrial areas of dogs in three groups were measured by echocardiography;the serum levels of renin, angiotensin Ⅱ(Ang Ⅱ), aldosterone(ALD), angiotensin Ⅱ type 1 receptor(AT1R), angiotensin Ⅱ type 2 receptor(AT2R) and angiotensin converting enzyme 2(ACE2) of the dogs in three groups were detected by ELISA method. Results:Compared with AF group,the atrial fibrillation burden of the dogs in SCS group was decresed obviously (P<0.05).The left and right atrial areas of dogs in AF group were larger than those in blank control group(P<0.05);the left atrial area of the dogs in SCS group was smaller than that in AF group (P<0.05),the right atrial area was also smaller,but there was no significant difference (P>0.05);the left and right atrial areas of the dogs in SCS group were larger than those in blank control group (P<0.05).The serum ACE2 level of dogs in AF group was lower than that in blank control group (P<0.05)and the levels of serum renin, Ang Ⅱ, ALD, AT1R and AT2R of the dogs in AF group were all higher than those in blank control group(P<0.05);the serum ACE2 level of the dogs in SCS group was higher than that in AF group(P<0.05)and the levels of serum renin, Ang Ⅱ, ALD, AT1R, and AT2R of the dogs in SCS group were all lower than those in AF group(P<0.05).Compared with before modeling,the serum ACE2 level of dogs in AF group after modeling was decresed(P<0.05),and the levels of serum renin, Ang Ⅱ, ALD, AT1R, and AT2R were incresed(P<0.05); compared with before treatment,the serum ACE2 level of the dogs in SCS group after treatment was incresed (P<0.05), and the levels of serum renin, Ang Ⅱ, ALD, AT1R, and AT2R were decresed (P<0.05). Conclusion:Atrial fibrillation can activate the RAAS. SCS therapy can suppress the atrial fibrillation and it can suppress RAAS activated by atrial fibrillation,so SCS can restraint the deterioration of atrial fibrillation.The serum RAAS indexes are so important observation indexes to evaluate the outcome of SCS in the treatment of atrial fibrillation.

Objective:To observe the long-term toxicity of chelerythrine on the morphology of lung tissue and the expression level of nuclear factor-kappa B (NF-κB) in lung tissue of the rats, and to investigate the related mechanism which causing the lung tissue damage of rats. Methods:A total of 120 Wistar rats were divided into control group(given normal saline) and low (3.7 mg·kg^-1), moderate (5.6 mg·kg^-1), high (8.4 mg·kg^-1) dosages of chelerythrine groups (n=30).The general condition of rats in various groups was observed;HE staining was used to observe the morphology of lung tissue of the rats in various groups;the serum interleukin-6 (IL-6),interleukin-8(IL-8) and tumor necrosis factor-α (TNF-α) levels of the rats in various groups were measured by ELISA method;real-time fluorescence quantitative PCR and Western blotting methods were used to detect the mRNA and protein levels of NF-κB and intercellular adhesion molecule-1(ICAM-1). Results:The accumulative mortality of the rats in high dosage of chelerythrine group was the highest, followed by moderate and low dosages of chelerythrine groups. The body weights and food intakes of the rats in different dosages of chelerythrine groups were significantly lower than that in control group(P<0.05), while the body weight and food intake of the rats in high dosage of chelerythrine group were lower than those in low and moderate dosages of chelerythrine groups (P<0.05).Chelerythrine led to pulmonary congestion and bloody ascites of the rats.The HE staining results showed that the injuries of lung tissue in different dosages of chelerythrine groups were aggravated with the increasing of the dosage of chelerythrine. Compared with control group, the levels of IL-6, IL-8, and TNF-α in serum of the rats in different dosages of chelerythrine groups were increased significantly (P<0.05); compared with low and moderate dosages of chelerythrine groups, the levels of IL-6, IL-8, and TNF-α in serum of the rats in high dosage of chelerythrine group were increased significantly (P<0.05). Compared with control group, the expression levels of NF-κB and ICAM-1 mRNA and proteins in lung tissue of the the rats in different dosages of chelerythrine groups were increased significantly (P<0.05)in a dose-dependent manner; compared with low and moderate dosages of chelerythrine groups, the expression levels of NF-κB and ICAM-1 mRNA and proteins in lung tissue of the rats in high dosage of chelerythrine group were increased(P<0.05). Conclusion:Chelerythrine has a long-term toxic effect on lung tissue in a dose-dependent manner of the rats. Moderate and high dosages of chelerythrine may aggravate the toxic lung injury through activation of NF-κB and production of inflammatory cytokines.

Objective:To observe the toxic effects of cockle shell-derived calcium carbonate nanoparticles (CCNPs) on the SD rats, and to preliminarily explore the safe dose and biological effects of CCNPs. Methods:A total of 40 healthy male SD rats were randomly divided into control group, low, middle and high doses of CCNPs groups (given 0, 30, 60, and 120 mg·kg^-1 CCNPs). The injection was consecutively given through the lateral tail vein once per day for 14 d. The body weights and daily feed intakes of the rats in various groups were measured. The haematology indexes, serum biochemical indexes, relative weights of organs and histopathology of main organs of the rats in various groups were detected. Results:In the 14-day repeated dose toxicity experiment, two rats died in high dose of CCNPs group, while no rats died in low and middle doses of CCNPs groups. The body weights and daily average intakes of the rats in different doses of CCNPs groups were slightly lower than those in control group, but the differences were not statistically significant (P>0.05). There were no significant differences in the serum indexes of the rats between different doses of CCNPs groups and control group(P>0.05). Compared with control group, the relative weights of lung of the rats in different doses of CCNPs groups were increased significant (P<0.05); the relative weights of spleen of the rats in low and middle doses of CCNPs groups were increased significantly (P<0.05); while the relative weight of spleen of the rats in high dose of CCNPs group had no significant difference(P>0.05). The histopathological analysis results showed that the mild inflammatory infiltration was found in some organs of the rats in low dose of CCNPs group, while no obviously pathological changes were found. The obviously histopathological changes were observed in the main organs of the rats in middle and high doses of CCNPs groups, including inflammatory cell infiltration, slight disorder of hepatocytes arrangement, increased vacuoles in hepatocytes; thickening of alveolar interstitium and formation of granulomas in the lung tissue; swelling, rupture and even necrosis of cardiac muscle fibers; atrophy and fissures of glomerulus and so on. Conclusion:Intravenous administration of low dose of CCNPs couldn't cause sever acute toxicity reaction.

Objective:To observe the inhibitory effect of tanshinone ⅡA (Tan ⅡA) on the proliferation of liver cancer HepG2 cells and its inductive effect on the migration and apoptosis of HepG2 cells, and to explore the possible mechanism. Methods:The human liver cancer HepG2 cells were cultured in vitro. The HepG2 cells were divided into blank group and different concentrations of Tan ⅡA groups. The cells in different concentrations of Tan ⅡA groups were added with Tan ⅡA at the final concentrations of 0.5, 1.0, 2.0, 5.0 and 10.0 mg·L^-1 Tan ⅡA and cultured for 24 h.The morphology of HepG2 cells in various groups was observed under inverted microscope. The inhibitory rates of proliferation of HepG2 cells in various groups were detected by MTT assay. The migration of HepG2 cells in various groups were evaluated by cell scratch assay. The expression levels of nuclear factor-κB (NF-κB) and matrix metalloproteinase-9 (MMP-9) mRNA were detected by RT-PCR method. Flow cytometry was used to detect the percentages of HepG2 cells at different cell cycles in various groups,and the apoptotic rates of HepG2 cells in various groups were detected by TUNEL method. Results:The morphology of HepG2 cells in different concentrations of Tan ⅡA groups were changed. Compared with blank group, the cells in 1.0 and 2.0 Tan ⅡA groups showed shrinkage,scattered connection and poor growth,and the inhibitory rates of proliferation of HepG2 cells in 1.0, 2.0, 5.0, and 10.0 mg·L^-1 Tan ⅡA groups were significantly increased (P<0.05) in a dose-dependent manner.The cell scratch assay results showed that with the increasing of Tan ⅡA concentration, the migration number of the cells in 2.0 and 5.0 mg·L^-1 Tan ⅡA groups were decreased significantly. Compared with 0.5 mg·L^-1 TanⅡA group, the expression levels of MMP-9 and NF-κB mRNA in the HepG2 cells in 1.0,2.0,and 5.0 mg·L^-1 Tan ⅡA groups were decresed(P<0.05 or P<0.01). The flow cytometry results showed that compared with blank group, the percentages of HepG2 cells in S phase in 1.0,2.0,5.0, and 10.0 mg·L^-1 Tan ⅡA groups were decreased (P<0.05),and the percentages of HepG2 cells in G₀/G₁ and G₂ phases were increased(P<0.05); the TUNEL results showed that compared with blank group, the apoptotic rates of HepG2 cells in 0.5, 1.0, 2.0, 5.0, and 10.0 mg·L^-1 Tan ⅡA groups werer increased (P<0.05 or P<0.01). Conclusion:Different concentrations of Tan ⅡA could significantly inhibit the proliferation and migration of human liver cancer HepG2 cells, and induce the apoptosis in a dose-dependent manner; its mechnasim may be related to the inhibition of the expressions of NF-κB and MMP-9 mRNA.

Objective:To investigate the effects of spina date seed (SDS)and albizzia julibrissin flower(AJF) water extracts on the hypothalamic-pituitary-adrenal (HPA) axis and inflammatory factors(IF) in the anxiety depression model rats. Methods:A total of 80 Wistar rats were randomly divided into blank group(n=10),model group(n=10), low dose of SDS group(n=10),high dose of SDS group(n=10), low dose of AJF group(n=10),high dose of AJF group(n=10),low dose of SDS-AJF group(n=10),and high dose of SDS-AJF group(n=10).The rat models of anxiety depression were induced by chronic restraint stress,solitary cage and corticosterone(CORT) subcutaneous injection.The forced swimming test (FST) was used to observe the immobility time of forced swimming of the rats in various groups;Elevated plus maze test (EPM) was used to observe the open arm entry(OE),close arm entry(CE),open arm entry time(OT), and close arm entry time(CT) of the rats in various groups.The total entry of open arms and close arms(TE),OE% and OT% were calculated.ELISA method was used to detect the levels of serum corticotropin releasing hormone (CRH), adrenocorticotropic hormone (ACTH),CORT,inflammatory factors interleukin-1β(IL-β) and interleukin-6 (IL-6) of the rats in various groups.HE staining was used to observe the morphology of hippocampal neurons of the rats in various groups. Results:Compared with model group,the immobility time of forced swimming of the rats in low dose of SDS-AJF group,high dose of SDS-AJF group,high dose of SDS group and high dose of AJF group was decreased (P<0.05 or P<0.01);compared with low dose of SDS group and low dose of AJF group, the immobility time of forced swimming of the rats in low dose of SDS-AJF group was shortened (P<0.05 or P<0.01);compared with high dose of SDS group and high dose of AJF group,the immobility time of forced swimming of the rats in high dose of SDS-AJF group was shortened(P<0.05 or P<0.01). Compared with model group,the OE% and OT% of the rats in low dose of SDS-AJF group,high dose of SDS-AJF group,high dose of SDS group and high dose of AJF group were increased (P<0.05 or P<0.01);compared with low dose of SDS group and low dose of AJF group,the OE% and OT% of the rats in low dose of SDS-AJF group were increased(P<0.05 or P<0.01);compared with high dose of SDS group and high dose of AJF group,the OE% and OT% of the rats in high dose of SDS-AJF group were increased (P<0.05 or P<0.01).Compared with model group,the levels of serum CRH, ACTH, CORT, IL-1β, and IL-6 of the rats in low dose of SDS-AJF group,high dose of SDS-AJF group,high dose of SDS group and high dose of AJF group were decreased(P<0.05 or P<0.01);compared with low dose of SDS group and low dose of AJF group,the levels of serum CRH, ACTH, CORT, IL-1β and IL-6 of the rats in low dose of SDS-AJF group was decreased(P<0.05 or P<0.01);compared with high dose of SDS group and high dose of AJF group,the levels of serum CRH, ACTH, CORT, IL-1β, and IL-6 of the rats in high dose of SDS-AJF group was decreased(P<0.05 or P<0.01).The HE staining results showed that the hippocampal neurons of the rats in model group were loosely arranged, and a large number of nerve cells were degenerated and the cell layer was reduced. The degeneration of hippocampal neurons of the rats in low and high doses of SDS-AJF groups were significantly relieved, and the nerve cells were arranged closely and the cell layer was clear. Conclusion:Combination of SDS and AJF can improve the behavior of the rats with anxiety depression,protect the hippocampal neurons of rats, reduce the inflammatory response, and adjust the disorder of HPA axis.

Objective:To investigate the therapeutic effects of YiXuanNing Granule on the ischemic vertigo of the mice and rats, and to clarify its effects on the inflammation damage and oxidative stress in brain tissue of the mice and rats. Methods:The vertigo models of mice and rats were established by mechanical vertigo and ligation of bilateral common carotid arteries(CCA),respectively. A total of 60 mice and 60 rats were selected and randomly divided into normal control group, model group, low dose of YiXuanNing Granule group (2.25 g·kg^-1/1.50 g·kg^-1), middle dose of YiXuanNing Granule group (4.50 g·kg^-1/3.00 g·kg^-1), high dose of YiXuanNing Granule group (9.00 g·kg^-1/6.00 g·kg^-1) and positive control group (1.50 g·kg^-1/1.00 mg·kg ^-1 flunarizine hydrochloride capsule),and there were 10 animals in each group. The mice and the rats in control group and model group were intragastrically administered with 10 mL·kg^-1 normal saline, and the mice and the rats in the other groups were given 10 mL·kg^-1 corresponding drug once a day for 7 d. The escape time of jumping off the platform of the mice and the rats in various groups were determined;the food intake of the mice in various groups were detected; the activities of superoxide dismutase (SOD) and the levels of malondialdehyde (MDA) in brain tissue of the rats in various groups were detected by colorimetric method. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in brain tissue of the rats in various groups were detected by ELISA method. Results:Compared with model group, the escape time of jumping off the platform of the mice and the rats in different doses of YiXuanNing Granule groups and positive control group was shortened (P<0.05 or P<0.01),and the food intake of the mice in various groups was increased. Compared with model group, the SOD activities in brain tissue of the rats in different doses of YiXuanNing Granule groups were significantly increased (P<0.05 or P<0.01); the MDA levels in brain tissue of the rats in different doses of YiXuanNing Granule groups were significantly decreased (P<0.01);compared with model group, the levels of TNF-α and IL-1β in brain tissue of the rats and the mice in middle and high doses of YiXuanNing Granule groups were significantly decreased (P<0.05 or P<0.01). Conclusion:YiXuanNing Granule can inhibit the inflammatory damage in brain tissue of the rats with ischemic vertigo and reduce the oxidative stress, thereby,it can play a therapeutic role in ischemic vertigo.

Objective:To explore the effects of eriC^F1 and eriC^F2 genes under the condition of different fluoride concentrations on the fluoride resistance of Streptococcus mutans(S.mutans) fluoride-resistant strain UA159-FR, and to elucidate the relationship between the differential expressions of eriC^F genes and the fluoride resistance of S.mutans fluoride-resistant strain. Methods:The recombinant E.coli competent cells BL21 were divided into eriC^F1 group (eriC^F1 + Peasy-Blunt E2), eriC^F2 group (eriC^F2 + Peasy-Blunt E2) and KZ group (Peasy-Blunt E2). After gene cloning and IPTG induction, the expression levels of EriC^F1 and EriC^F2 proteins in three groups were detected, and the concentrations of bacterial solution and the growth rates of bacterium of strains in three groups were determined under different concentrations of fluoride conditions (1.5,2.0,2.5,and 3.0 g·L^-1 sodium fluoride);the growth curves were drawn. Results:At logarithmic phase(8 h) and stable phase(16 h), the final concentrations of bacterial solution and the growth rates of the strains in three groups under different concentrations of fluorine environments (1.5,2.0,2.5 and 3.0 g·L^-1 sodium fluoride) were compared in the same group,and the final concentrations and the growth rates of bacterial solution of the strains in three groups were decreased with the increasing of sodium fluoride concentration; the final concentrations and the growth rates of bacterial solution results were eriC^F2 group > eriC^F1 group > KZ group (P<0.05 or P<0.01).compared between three groups under the same concentration of sodium fluoride, the concentration of bacterial solution in eriC^F2 group was increased significantly compared with eriC^F1 group(P<0.01). Conclusion:Both eriC^F1 and eriC^F2 genes can enhance the fluoride resistance of E.coli, and the fluoride resistance of E.coli expressing eriC^F2 gene is stronger than that of E.coli expressing eriC^F1 gene.

Objective:To observe the effects of RuanJianXiaoYing Granule on the expressions of specific transcription factors and cytokines of Treg and Th17 in the Hashimoto thyroiditis(HT) rats with liver depression and spleen deficiency, and to investigate the therapeutic effect of RuanJianXiaoYing Granule on HT and its immunological mechanism. Methods:A total of 48 SD rats were randomly divided into normal group (12 rats) and modeling group (36 rats).The HT model was established by subcutaneous injection of excessive iodine drinking water and thyroglobulin,and then combined with chronic restraint stress, excessive fatigue and dietary disorders to prepare the rat models with liver depression and spleen deficiency.After modeling,the rats were randomly divided into model group(n=12),Tripterygium wilfordii group(n=12), and RuanJianXiaoYing group(n=12),and the rats were treated for 8 weeks. In addition to the dead rats and unqualified specimens during the experiment,the test results of 10 rats in each group were retained.The general situation of the rats in various groups was observed; the abdominal aortic blood and thyroid gland tissue of the rats in various groups were collected after the last administration;the levels of thyroglobulin antibody(TGAb), thyroid peroxidase antibody(TPOAb),free triiodothyronine(FT3), free thyroxine(FT4), thyrotropin-releasing hormone(TRH) and thyrotropin(TSH) in serum and the levels of IL-17, IL-23 and IL-10 were detected by ELISA method;the pathomorphology of thyroid gland tissue of the rats in various groups were detected by HE staining; the expression levels of Foxp3 and RORγt proteins in thyroid gland tissue of the rats in various groups were detected by Western blotting method. Results:Compared with normal group,the levels of serum TGAb and TPOAb of the rats in model group, Tripterygium wilfordii group and RuanJianXiaoYing~group were increased (P< 0.05);the levels of FT3, FT4 and TRH were decreased(P<0.05),the levels of TSH were increased(P<0.05);the expression levels of Foxp3 protein in thyroid gland tissue were decreased(P<0.05), and the expression levels of RORγt protein were increased (P<0.05);the levels of serum IL-10 were decreased(P<0.05),and the levels of serum IL-17 and IL-23 were increased(P<0.05).Compared with model group, the levels of TGAb and TPOAb of the rats in Tripterygium wilfordii group and RuanJianXiaoYing group were decreased(P<0.05);the levels of FT3, FT4 and TRH were increased (P< 0.05), and the levels of TSH were decreased (P< 0.05);the expression levels of Foxp3 protein in thyroid gland tissue were increased(P<0.05), and the expression levels of RORγt protein were decreased (P< 0.05);the levels of serum IL-10 were increased(P< 0.05), and the levels of IL -17 and IL-23 were decreased (P< 0.05).Compared with Tripterygium wilfordii group, the levels of TGAb, TPOAb,FT3,FT4,and TRH of the rats in RuanJianXiaoYing group were increased (P< 0.05),and the levels of TSH of the rats in RuanJianXiaoYing group was decreased(P<0.05);the expression level of Foxp3 protein in thyroid gland tissue of the rats was increased, and the expression level of RORγt protein in thyroid gland tissue was decreased,but there no were significant differences(P>0.05);the level of serum IL-10 was increased(P<0.05),and the levels of serum IL-17 and IL-23 were decreased(P<0.05). The follicular cavity of thyroid gland in normal group had regular morphology, complete structure and no lymphocyte infiltration. In model group, the follicular cavity of thyroid gland was enlarged,the follicular structure was destroyed, and a large number of lymphocyte infiltration was observed in and around the follicular cavity. In Tripterygium wilfordii group, the follicular cavity of thyroid gland was enlarged, part of the follicular structure was destroyed, and there was a small amount of lymphocyte infiltration in and around the follicular cavity, which was less than that in model group.The morphological changes of thyroid gland in RuanJianXiaoYing group were similar to those in Tripterygium wilfordii group. Conclusion:RuanJianXiaoYing Granule has the therapeutic effect in the HT rats with liver depression and spleen deficiency by regulating the expressions of the specific transcription factors Foxp3 and RORγt protein and related cytokines of Treg and Th17.

Objective:To investigate the changes of the levels of interleukin-1β (IL-1β),interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and the P38 mitogen-activated protein kinase(MAPK)/P65 nuclear transcription factor-κB (P65NF-κB) signaling pathways in the cerebral cortex tissue of the mice with heat stroke,and to elucidate its mechniasm. Methods:A total of 60 mice were randomly divided into control group, heat stroke 1 h group,heat stroke 6 h group, and heat stroke 24 h group(1, 6, and 24 h after leaving warehouse in the heat stroke model mice), and there were 15 mice in each group. HE staining was used to detect the morphology of cerebral cortexy tissue of the mice in various groups; the levels of IL-1β, IL-6 and TNF-α in cerebral cortex tissue of the mice in various groups were measured by ELISA method;the expression levels of P38MAPK, P65NF-κB, p-P38MAPK and p-P65NF-κB proteins in cerebral cortex tissue of the mice in various groups were determined by Western blotting method. Results:Compared with control group, the body mass loss of mice in heat stroke groups was increased (P<0.05),and the anal temperatures of the mice in heat stroke 1 h group and heat stroke 6 h group were significantly decreased(P<0.05).The HE staining results showed that the cerebral cortex tissue of the mice in control group were normal;in heat stroke 1 h group, a large number of brain cells were pyknosisd and stained darkly,the blood vessels were compressed and deformed, and a large number of edema was found outside the wall;in heat stroke 6 h group,the number of pyknosisd cells was decreased, and the extravascular edema was reduced;in heat stroke 24 h group,the pyknosisd brain cells and the extravascular edema were rare.Compared with control group, the levels of IL-1β, IL-6, TNF-α and the expression levels of p-P38MAPK and p-P65NF-κB proteins in cerebral cortex tissue of the mice in heat stroke 1 h group and 6 h group were increased (P<0.05). The levels of IL-1β, IL-6, TNF-α and the expression levels of p-P38MAPK and p-P65NF-κB proteins in cerebral cortex tissue of the mice in heat stroke 6 h group were lower than those in heat stroke 1 h group (P<0.05). Conclusion:The levels of inflammatory cytokines in cerebral cortex tissue of the mice with heat stroke are elevated,and the P38MAPK/P65NF-κB signaling pathway is activated. This inflammatory response of central nervous system may be related to the activation of signaling pathways.

Objective:To investigate the effect of nerve growth factor (NGF) on the expression level of growth differentiation factor-15(GDF-15) in the brain tissue of the rats with cerebral infarction, and to elucidate the mechanism of NGF in the rats with cerebral infarction. Methods:The middle cerebral artery occlusion (MCAO) models were established by the Longa's method. A total of 54 SD rats were randomly divided into sham operation group, model group and NGF group, and there were 18 rats in each group.The rats in NGF group were given NGF (50 μg·kg^-1) by intraperitoneal injection,and equal volume of normal saline was given to the rats in sham operation (the artery was isolated without ligation) group and model group. The neurological function scores of the rats in various groups were measured 1, 3 and 7 d after operation; HE staining was used to observe the pathomorphology of nerves in brain tissue and immunohistochemical staining was used to detect the number of GDF-15 positive cells; the expression levels of GDF-15 in brain tissue of the rats in various groups were detected by ELISA method. Results:The results of HE staining showed that the degrees nerve cell necrosis, interstitial edema and glial cell proliferation were relatively low in NGF group 7 d after operation. Compared with sham operation group, the neurological function scores, the number of GDF-15 positive cells,and the expression levels of GDF-15 in brain tissue of the rats in model group and NGF group were significantly increased (P<0.05). Compared with model group, the neurological function scores of the rats in NGF group were significantly decreased 3 and 7 d after operation (P<0.05), and the number of GDF-15 positive cells and the expression levels of GDF-15 in brain tissue of the rats in NGF group were significantly increased at different time points after operation (P<0.05). Conclusion:NGF can protect the brain nerve by up-regulating the expression level of GDF-15 in brain tissue and improving the nerve function.

Objective:To investigate the associations between single nucleotide polymorphisms (SNP) of integrin α-subunit 1 (ITGA1) gene and the risk of non-cardia gastric cancer, and to elucidate the roles of ITGA1 gene polymorphisms in the risk of non-cardia gastric cancer. Methods:The case-control study design was used. A total of 288 patients with non-cardia gastric cancer were selected as case group, and 281 community physical examinees were selected as control group in Han population in Boatou City in Inner Mongolia Autonomous Region. The SNP of rs1862610, rs2432143, and rs2447867 sites of subjects in two groups were genotyped by using TaqMan method,and the haplotype was constructed by Haploview software 4.0. The odds ratio (OR) and 95% confidence intervals (95%CI) were measured by conditional Logistic regression analysis to evaluate the correlations between the frequencies of allele, genotype, and haplotypes and the susceptibility to non-cardia gastric cancer. Results:The SNP of rs1862610, rs2432143, and rs2447867 sites in ITGA1 gene were not associated with the risk of non-cardia gastric cancer (P>0.05). The SNP of rs1862610 and rs2432143 sites were in strong linkage disequilibrium(D'=1) and formed a block. Compared with those who carried haplotype CT, those who carried haplotype CC had an increasing risk of non-cardiac gastric cancer(CC vs CT:OR=1.42, 95% CI:1.02-1.99). Conclusion:The ITGA1 gene polymorphisms may play a role in the risk of non-cardia gastric cancer in Han population in Baotou in Inner Mongolia Autonomous Region. The risk of non-cardia gastric cancer in the person who carries the SNP rs1862610-rs2432143 CC haplotype is increased.

Objective:To investigate the tracing effect of new nano-carbon(Kanalin)on the pelvic lymph nodes of the patients with bladder cancer during the laparoscopic radical cystectomy(RC), and to explain the significance of its application in the precision surgical treatment. Methods:A total of 44 patients with bladder cancer who underwent RC were divided into tracer group(n=22) and control group (n=22) according to whether Kanalin was used before operation or not. The laparoscopic RC combined with standard pelvic lymph node dissection(LND) was performed in the patients in two groups. The effectiveness evaluation indexes and safety evaluation indexes of the patients in two groups were collected and compared. The effectiveness evaluation indexes included black staining situation of lymph nodes during operation,the number of detected lymph nodes,the black staining rate of lymph nodes,the number of positive lymph nodes, and operation time. The safety evaluation indexes included the blood loss and the incidences of perioperative complications. Results:The black staining effect of lymph nodes of the patients in tracer group was definite and there were no tracer-related side effects and complications during the perioperative period. Compared with control group, the clearance rate and detection rate of lymph nodes of the patients in tracer group were increased (P<0.01). The black staining rate of lymph nodes of each case was higher than 50%, even up to 85%. There were no significant differences in the number of positive lymph nodes,the operation time, the blood loss and the perioperative complication rates of the patients between two groups(P>0.05). Conclusion:During the laparoscopic RC, the application of Kanalin for pelvic lymph node tracing is safe, without toxic and side effects;Kanalin is helpful for increasing the intraoperative clearance rates and postoperative pathological detection rates of lymph nodes.

Objective:To evaluate the practicability and feasibility of detection of circulating tumor cells (CTC) combined with exosome miR-21 in the diagnosis of ground glass opacity(GGO), and to provide the evidence for the diagnosis of GGO. Methods:Seventy patients were selected according to the diagnostic and exclusion criteria of GGO. All the patients underwent thoracoscopic segmentectomy. The patients diagnosed as lung cancer after operation were regarded as case group (n=24), and the patients diagnosed as benign lesion after operation were regarded as control group (n=46). Before operation, CTC was captured from the peripheral blood by negative enrichment of immunomagnetic beads. CTC was labeled with tumor-specific folate ligand-oligonucleotide conjugate. The exosomes of the same specimen were detected, purified and identified, and the expression levels of serum exosome miR-21 of the patients in two groups were detected. The diagnostic efficacies of CTC and exosome miR-21 in the GGO paitents were evaluated by ROC curves. Results:The positive rate of CTC in the patients in case group was higher than that in control group (P<0.05). There were no significant differences in the positive rates of CTC between the patients with different pathological types in case group and control group (P>0.05). The purified samples showed exosome characteristics. The expression level of serum exosome miR-21 in the patients in case group was higher than that in control group (P<0.05). The area under ROC curve (AUC) of CTC was 0.891, and the AUC of exosome miR-21 was 0.711, which showed good specificity. The sensitivity and specificity of the combined detection were 85.64% and 86.79%,respectively. Conclusion:The detection of CTC combined with exosome miR-21 can be used in the clinical diagnosis of early lung cancer with high sensitivity and specificity.

Objective:To investigate the relationship between the expression of glucose-regulated protein 78 (GRP78) and gemcitabine chemotherapy in the patients with non-small cell lung cancer (NSCLC) and the effects of GRP78 on the viability of lung adenocarcinoma SPCA1 cells and the chemosensitivity of gemcitabine, and to elucidate its mechanisms. Methods:The positive expression rates of GRP78 in 32 cases of cancer tissue of the NSCLC patients were detected by immunohistochemical staining. The SPCA1 cells with high expression of GRP78 were selected as the subjects. RNA interference technique was used to down-regulate the expression of GRP78 in SPCA1 cells(interference group) and the cells treated with shNC were used as control group. MTT assay was used to detect the viabilities of SPCA1 cells in various groups.Negative control group,interference group,control+gemcitabine group,and interference+gemcitabine group were set up; colone formation assay was used to detect the colone formation rates of SPCA1 cells in various groups; Western blotting method was used to detect the expression amount of Akt, p-Akt, PI3K, and p-PI3K in the SPCA1 cells in various groups. Results:The immunohistochemical staining results showed that the positive expression rate of GRP78 in cancer tissue in the remission NSCLC patients was significantly higher than that in the no-remission NSCLC patients after treated with gemcitabine (P<0.05). The MTT assay results showed that compared with negative control group, the viability of SPCA1 cells in interference group was decreased significantly (P<0.05),and the viability of SPCA1 cells in interference+gemcitabine group was significantly decresed(P<0.05). The Western blotting results showed that compared with negative control group, the expression amounts of p-Akt and p-PI3K in the SPCA1 cells in interference group were decreased, and the expression amounts of p-Akt and p-PI3K in the SPCA1 cells in interference+gemcitabine group were decreased significantly. Conclusion:Interference of GRP78 may increase the sensitivity of gemcitabine to chemotherapy,and GRP78 may reduce the sensitivity of NSCLC patients to gemcitabine through PI3K/Akt pathway.

Objective:To investigate the relationship between interleukin-6 (IL-6) and papillary thyroid carcinoma (PTC), and to elucidate the role of IL-6 in the occurrence and development of PTC and its possible mechanism. Methods:The serum levels of IL-6 were measured by ELISA method in 5 patients with PTC(case 1 group), 5 patients with Hashimoto thyroiditis (HT) complicated with PTC(case 2 group) and 5 normal persons(normal group). The thyroid tissues of 20 patients with PTC(PTC group), 15 patients with HT complicated with PTC(HT +PTC group) and 18 normal persons(control group) were collected. The positive expression rates of IL-6, IL-6 receptor (IL-6R), nuclear factor kappa-B(NF-κB) and RET/PTC proteins in thyroid tissue of the subjects in various groups were detected by immunohistochemistry(IHC) method; the relationships between IL-6, IL-6R and NF-κB, RET/PTC expressions were analyzed. Results:In the patients with PTC, the complex papillary hyperplasia of thyroid epithelial cells accompanying with ground glass like nuclei and nuclear sulcus and inclusions were seen; in the patients with HT complicated with PTC, in addition to the typical pathological changes of PTC, the diffuse lymphoid tissue infiltration accompanying with lymphoid follicle formation,and the coexistence of hyperplasia and atrophic thyroid follicular epithelium with eosinophic degeneration of thyroid epithelium and interstitial fibrosis were also found. The lobular structrure of normal thyroid tissue was found,and the follicular cells were in the same size The ELISA results showed that the serum IL-6 levels of the patients in PTC group and HT+ PTC group were higher than those in normal group (P<0.05);the IHC results showed that the positive expression rates of IL-6, IL-6R, NF-κB, and RET/PTC in thyroid tissue of the patients in PTC group were higher than those in control group (P<0.05). The expressions of IL-6,and IL-6R were positively correlated with the expressions of NF-κB and RET/PTC protein (P<0.05). Conclusion:IL-6 may be involved in the occurrence and development of PTC through the RET/PTC-NF-B signal pathway.

Objective:To observe the changes of T,B and NK lymphocyte subsets in peripheral blood of the patients with lung adenocarcinoma in the early stage after single-port thoracoscopic lobectomy, and to explore their relationships with the clinicopathological parameters of the patients and the possible clinical significances. Methods:A total of 66 patients with lung adenocarcinoma who underwent single-port thoracoscopic lobectomy were selected and the peripheral blood was respectively collected on the day of admission (D0),the 1st day (D1) and the 5th day (D5) after operation. The blood cell counter and flow cytometry were used to detect the percentages of CD4+T,CD8+T,CD19+B and CD56+NK lymphocytes. At the same time, the variations of cell percentages of T,B and NK lymphocytes (ΔD0-1,ΔD0-5 and ΔD1-5)before and after operation were calculated, respectively. The correlations between the changes of cell percentages (ΔD0-1,ΔD0-5 and ΔD1-5)of the patients in the early stage after operation and the clinicopathological parameters were analyzed. Results:Compared with D0, the percentages of CD4+T,CD8+T,CD19+B, and CD56+NK lymphocytes in peripheral blood of the patients on D1 and D5 were significantly decreased (P<0.01).The percentages of CD4+T and CD8+T lymphocytes on D5 were significantly higher than those on D1 (t =5.362, P<0.01;t=5.202,P<0.05). The percentages of CD19+B and CD56+NK lymphocytes showed a decreasing trend, but there were no significant differences in the percentages of CD19+B and CD56+NK lymphocytes of the patients between D5 and D1 (P>0.05). The percentage of CD8+ T cells in smokers was lower than that in non-smokers on D0 and D5 (t=2.162, P<0.05;t=1.974,P<0.05).The percentage of CD19+ B lymphocytes in smokers was higher than that in the non-smokers on D0 (t=-1.137,P<0.05). The percentage of CD8+ T lymphocytes in stage T1 was higher than that in stage T2-3 on D0 (t=1.684, P<0.05).The percentage of CD19+ B lymphocytes in stage T1 was higher than that in stage T2-3 (t=1.431, P<0.05), which was in stage Ⅰ higher than that in stage Ⅱ-Ⅲ on D5 (t=2.134, P<0.05).The variation in CD4 + T lymphocyte percentage of the male patients (ΔD1-5) was higher than that of the female patients (t=1.707,P<0.05).The variations in CD8+ T lymphocyte percentage (ΔD0-1 and ΔD0-5) had negative relationship with T stage(r=-0.366,P<0.01;r=-0.252,P<0.05). The variations in CD19+ B lymphocyte percentages (ΔD0-5 and ΔD1-5) of the patients in stage Ⅱ-Ⅲ were higher than those in stage Ⅰ (t=2.075,P<0.05;t=2.140,P<0.05). The variation of cell percentages of CD56+ NK lymphocytes(ΔD0-5) had significantly relationships with the age and smoking status of the patients.The ΔD0-5 in the older patients (≥ 60 years) was lower than that in the younger patients (< 60 years) and the ΔD0-5 in the smokers was lower than those in the non-smokers (t=1.557,P<0.05).The variation of lymphocyte percentage of CD56+ NK (ΔD1-5) had negative relationship with T stage (r=-0.592,P<0.05). Conclusion:The change degrees of percentages of lymphocyte subsets are correlated with the patients' age, gender, smoking status, and T stage.

Objective:To study the application of individualized semi-shoulder prosthesis under the assistance of 3D printing technology in the shoulder replacement surgery of one patient with bone defect caused by a complex proximal humerus fracture internal fixation failure, and to provide the basis for the repairment and reconstruction of articular bone defect. Methods:One patient was admitted to the hospital because of the internal fixation failure after open reduction and internal fixation for left proximal humeral fractures and proximal collapse and necrosis of humerus. The humerus CT data of this patient was collected and the three-dimensional reconstruction was conducted. The models of stereo lithography apparatus resin on healthy side and affected side of this patient were manufactured by 3D printing technology,and the length of osteotomy was designed preoperatively according to the bone destruction condition. The shape of customized prosthesis was designed on the basis of mirror image of the contralateral humerus head. The Ti₆Al₄V semi-shoulder prosthesis was manufactured by the means of Electron Beam Melting technology. The preoperative simulative operation was carried out,and the matching experiment of the customized prosthesis and residual humerus model was performed; the humerus osteotomy was performed in accordance with the preoperative design in operation. The customized prosthesis was implanted to the designed location. The patients were followed up at 1, 3, 6,and 12 months after operation. Results:The operation time was 150 min. The intraoperative blood loss was 290 mL. The postoperative humerus X-ray examination at 1, 3, 6,and 12 months after operation were performed and the results showed that the prosthesis was observed in good position without loosening. There were also no infection (ESR and C-reactive protein level were normal) as well as dislocation,nerve damage and periprosthetic fractures,etc. The UCLA scores were 13 points before and during operation,15 points at 1 month after operation,22 points at 3 months after operation, 28 points at 6 months after operation,and 30 points at 12 months after operation. The Constant-Murley scores were 21 points before and during operation,28 points at 1 month after operation,61 points at 3 months after operation, 65 points at 6 months after operation, and 70 points at 12 months after operation. The Disbility of Arm shoulder and Hand(DASH) scores were 100 points before and during operation,63 points at 1 month after operation,50 points at 3 months after operation, 44 points at 6 months after operation, 25 points at 12 months after operation. The rehabilitation of daily life ability of the affected shoulder of the patient after operation was satisfied. Conclusion:Individualized semi-shoulder joint prosthesis under assistance of 3D printing technology can achieve the perfect anatomy reconstruction and partial function restoration of shoulder joint and the curative effect of short-term follow-up of the patient isobvious.

Objective:To explore the expressions of Foxp3⁺ Tregs and CD11c⁺ mDCs in the colorectal cancer (CRC) tissue and their infiltrations in the tumor draining lymph node (TDLN) tissue,and to explore the clinical significances. Methods:Fifty-two samples of surgical resection of the CRC patients were collected. The expressions of Foxp3⁺ Tregs and CD11c⁺ mDCs in the specimens of the CRC patients and TDLN tissue were detected by immunohistochemistry, and the relationship between the expression frequencies of Foxp3⁺ Tregs and CD11c⁺ mDCs and the clinicopathological characteristics of the patients were discussed. Results:The positive expression frequency of Foxp3⁺ Tregs in cancer tissue of the CRC patients was higher than that in adjacent normal mucosa tissue(P<0.01); The expression frequency of Foxp3⁺ Tregs in cancer tissue in the patients with positive lymph node metastasis was higher than that in the patients with negative lymph node metastasis(P<0.01); the expression frequency of Foxp3⁺ Tregs in the patients in TNM stage Ⅲ+Ⅳ was higher than that in TNM stage Ⅰ+Ⅱ(P<0.01); the positive expression frequency of Foxp3⁺ Tregs in metastatic TDLN (mTDLN) tissue was higher than that in metastatic free TDLN (mfTDLN) tissue(P<0.01). The positive expression frequency of CD11c⁺ mDCs in cancer tissue of the CRC patients was higher than that in adjacent normal mucosa tissue(P<0.01); the positive expression frequency of CD11c⁺ mDCs in the patients with positive lymph node metastasis was lower than that in the patients with negative lymph node metastasis(P<0.01);the positive expression frequency of CD11c⁺ mDCs in the patients in TNM stage Ⅲ+Ⅳ was lower than that in the patients in TNM stage Ⅰ+Ⅱ(P<0.01); the positive expression frequencies of CD11c⁺ mDCs in cancer tissue with different depths invasion were significantly different (P<0.05);the positive expression frequency of CD11c⁺ mDCs in mTDLN tissue was lower than that in mfTDLN tissue (P<0.01).There were no significant correlations between the expressions of Foxp3⁺ Tregs and CD11c⁺ mDCs in the CRC and TDLN tissues(P>0.05). Conclusion:The occurrence and development of CRC are related to the changes of Foxp3⁺ Tregs and CD11c⁺ mDCs expression frequencies in the local microenvironment of cancer tissue.Foxp3⁺ Tregs and CD11c⁺ mDCs play a role in inhibiting the cellular immunity in tumor microenvironment.

Objective:To analyze the facial profile of the patients with congenitally missing 2 mandibular incisors, and to investigate the changes of facial profile of the patients before and after orthodontic treatment by extraction of 2 maxillary premolars. Methods:The cephalometrics of 64 patients with congenitally missing 2 mandibular incisors were collected. The initial cephalometrics of 64 patients were used to characterize the facial profile. A total of 38 cases among all the 64 patients underwent the orthodontic treatment by extraction of 2 maxillary premolars, and the initial and final cephalometrics were used to compare the changes of facial profile before and after treatment. The cephalometric measurement (including hard tissue and soft tissue items) was performed and the results were analyzed statistically. Results:Compared with the normal values, the overjet, overbite, ANB, U1-NA angle, face angle, Y axis, Pg-NB, face convexity, soft tissue facial angle, H angle, and mentolabial sulcus to H-Line in the patients with congenitally missing 2 mandibular incisors were increased; SNB, L1-NB angle, L1-NB distance, L1-MP, occlusal plane angle, mentolabial sulcus anglewere decreased, and the differences were statistically significant (P<0.05). After orthodontic extraction treatment, the overjet, overbite, U1-NA angle, U1-NA distance, U1-SN, H angle,and subnasale to H-Line of the patients were decreased; the L1-NB angle, L1-NB distance, L1-MP, occlusal plane angle, Pg-NB, and mentolabial sulcus angle were increased; the differences were statistically significant (P<0.05). Conclusion:The mandibular development is insufficient but the chin development is adequate in the patients with congenitally missing 2 mandibular incisors, with larger overjet and overbite, lingually inclined mandibular incisors, deeper and sharper mentolabial sulcus. After treatment by extraction of 2 maxillary premolars, the overall appearance of the lateral profile of the patients remains unchanged, but the occlusion of anterior teeth is significantly improved.

Objective:To detect the expressions of periostin, matrix metalloproteinase-8 (MMP-8)and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2)in gingival tissue of the patients with chronic periodontitis,and to explore the possible role of periostin in the occurrence and development of chronic periodontitis and its related biological mechanism. Methods:A total of 23 normal gingival tissues(control group) and 19 inflammatory gingival tissues (case group)were selected. The expressions of periostin, MMP-8 and TIMP-2 in gingival tissue of the patients in two groups were detected by immunohistochemical SP method.The correlation between the expressions of periositn and MMP-8 and TIMP-2 in gingival tissue of the patients in two groups were detected by Spearman analysis. Results:The positive expression rate of periostin in gingival tissue of the patients in case group was lower than that in control group (P<0.05). The correlative analysis results demonstrated that the expression of periostin had negative relationship with the expression of MMP-8 (r=-0.51, P<0.05) in control group, and positive relationship with the expression of TIMP-2 (r=0.50, P<0.05);in case group, the expression of periostin had negative relationship with the expression of MMP-8 (r=-0.482, P<0.05), and positive relationship with the expression of TIMP-2 (r=0.655, P<0.05). Conclusion:Periostin can be used as a preliminary indicator for maintaining the structural stability of gingival tissue, and become a research target for periodontal tissue regeneration.

Objective:To study the values of exfoliated cell smear, DNA ploidy analysis, cell block and their combined detection in the diagnosis of malignant pleural effusion, and to provide the evidence for the diagnosis and treatment of malignant pleural effusion. Methods:A total of 300 cases of pleural effusion specimens were analyzed by DNA ploidy analysis to judge the benign and malignant pleural effusion;the centrifuged cell pellet was used for smear, the cell block was made, and the source of malignant pleural effusion was judged by immunohistochemistry method. The sensitivities and specificities of three methods and their combined detection in the diagnosis of malignant pleural effusion were compared. Results:The sensitivities of exfoliated cell smears, DNA ploidy analysis and cell block in the diagnosis of malignant pleural effusion were 83.13%, 84.44%, and 79.52%, respectively; the specificities were 82.95%, 86.64%, and 83.87%, respectively; the sensitivity of parallel test in the diagnosis of malignant pleural effusion was 98.79%; the specificity of serial test in the diagnosis of malignant pleural effusion was 99.54%. Conclusion:The combined detection of three methods can significantly improve the clinical diagnotic effect of malignant pleural effusion compared with single detection of three methods.

Objective:To investigate the effects of adiponectin (APN,also known as ADIPOQ)-related miR-371b-5p with differential expressions in the epicardial adipose tissue (EAT) and plasma in the patients with coronary artery disease (CAD) on the secretions of APN, Kruppel-like factor 4 (KLF4), interleukin-6 (IL-6) and monocyte chemokine 1 (MCP-1), and to elucidate the mechanism of action of miR-371b-5p in CAD. Methods:The EAT and blood samples were collected from the patients with CAD (CAD group,n=34) and non-CAD (non-CAD group,n=16). Real-time quantitative PCR (qRT-PCR) was used to detect the expression levels of miR-371b-5p in the EAT and plasma of the patients in two groups. The successfully induced 3T3-L1 preadipocytes were divided into(without any treatment), miR-371b-5p over-expression group (transfected with miR-371b-5p mimics) and miR-371b-5p inhibitor group (transfected with miR-371b-5p inhibitor). The relative expression levels of APN, KLF4, IL-6 and MCP-1 mRNA in the 3T3-L1 preadipocytes were detected by qRT-PCR method. The miR-371b-5p was analyzed by bioinformatics. Results:The expression levels of miR-371b-5p in the EAT and plasma of the patients in CAD group were higher than those in non-CAD group(P<0.01). Compared with blank control group,the expression levels of APN and KLF4 mRNA in 3T3-L1 preadipocytes in miR-371b-5p over-expression group were decreased(P=0.043, P=0.045), and the expression levels of IL-6 and MCP-1 mRNA in 3T3-L1 preadipocytes in miR-371b-5p over-expression group were increased (P=0.037, P=0.041). Compared with blank control group, the expression levels of APN and KLF4 in 3T3-L1 preadipocytes in miR-371b-5p inhibitor group were increased(P=0.025, P=0.027), and the expression levels of IL-6 and MCP-1 mRNA in 3T3-L1 preadipocytes in miR-371b-5p inhibitor group were decreased (P=0.039, P=0.041). The target genes predicted by miR-371b-5p were enriched in multiple biological functions and processes. The KEGG pathway was mainly enriched in the signaling pathways such as adipocytokines. The protein interaction results showed that the target genes APN,leptin(LEP) and AKT1 encoded proteins predicted by miR-371b-5p were located at the core of the interaction network. Conclusion:The expression levels of anti-inflammatory factors in the cells with over-expression of miR-371b-5p in EAT of the patients with CAD are decreased, the expression levels of inflammatory factors are increased,and miR-371b-5p is expected to become a new target for CAD treatment.

Objective:To analyze the results of bacteria distribution of wound secretion and the drug susceptibility test of the infected patients from Department of Hand Surgery,and to provide the evidence for antibiotic selection in the infected patients in early stage of treatment. Methods:The clincal data of hospitalized patients from Department of Hand Surgery were collected.The patients were divided into three groups according to the wound conditions. The patients in Group 1 who had no wound while being in hospital were infected after operation;the patients in Group 2 had the contaminated wound but no infection while being in hospital;the patients in Group 3 had definite infection while being in hospital. The results of bacterium culture of wound secretion and drug susceptibility test of the patients in three groups were analyzed. Results:The clinical data of 297 patients were obtained, including 17 patients from Group 1, 201 patients from Group 2, and 79 patients from Group 3. A total of 406 strains of 54 species of bacteria were isolated from 297 patients,including 178 strains(43.84%,178/406) gram-positive bacteria and 228 strains(56.2%,228/406) gram-negative bacteria. The four most common strains were Staphylococcus aureus(15.8%,64/406), Staphylococcus epidermidis(14.0%,57/406),Enterobacter cloacae(10.3%,42/406),and Serratia marcescens(9.9%,40/406).The results of drug susceptibility test showed that the detection rates of methicillin-resistant staphylococcus aureus(MRSA) and staphylococcus epidermidis were 9.4%(6/64) and 72.0%(46/64), respectively. There were no vancomycin-resistant, linezolid-resistant, and tigecycline-resistant positive strains among Staphylococcus susceptibility;and there were no the carbapenem-resistant positive strains among Enterobacter cloacae and Serratia marcescens.The susceptibility rate of Enterobacter cloacae to ceftriaxone was 83.3%(35/42), and its susceptibility to ceftazidime was 90.5%(38/42). The susceptibility rate of Serratia marcescens to ceftriaxone was 100.0%(40/40), and its susceptibility to ceftazidime was 97.5%(39/40). Conclusion:The first generation of cephalosporin and penicillinase-stabilized penicillin can be used as the preferred empirical antibiotics for the infected patients from Deparment of Hand Surgery in our hospital.The third and fourth generation cephalosporins and quinolones antibiotics can be used as the preferred antibiotics for the infected patients with open trauma as well as the possibility of G-bacillus infection.

Objective:To compare the urinary neutrophil gelatinase-associated lipocalin (NGAL) levels in the acute kidney injury (AKI) and non-AKI premature infants,and to explore the clinical value of NGAL level in the early diagnosis of AKI. Methods:A total of 85 premature infants were divided into AKI group (n=30) and non-AKI group (n=55) according to whether AKI occurred.The levels of urinary NGAL were detected by enzyme-linked immunosorbent assay (ELISA)method at 24, 48, 72, 120, and 168 h after birth. The levels of serum creatinine (Scr) of preterm infants were detected by dry chemical method.The receiver operation cllaracteristics(ROC) curve is used to judge the diagnostic value of each indicator. Results:There were no significant differences in gender, gestational ages, body weights at birth, ages at NICU of the premature infants between two groups (P>0.05).There were significant differences in the urinary NGAL levels of the premature infants between two groups at different time points (F=62.710, P<0.01);the urinary NGAL level was increased at 24 h after birth, reached the highest peak at 72 h afger birth, and decreased from 72 h after birth.The urinary NGAL levels of the premature infants in AKI group were higher than those in non-AKI group at 24, 48, 72 and 120 h after birth (P<0.05).There were statistically significant differences in the Scr levels of the pretmature infants between two groups at different time points (F=27.332, P<0.05).The Scr levels of the premature infants in AKI group were higher than those in non-AKI group at 48,72 and 120 h after birth (P<0.05). The area under ROC curve (AUC) of urinary NGAL level at 24 after birth was significantly greater than that of Scr level (P<0.01).When the cutoff value of urinary NGAL level was 6.95 ng·dL^-1,the sensitivity and specificity of urinary NGAL level in diagnosis of AKI were 0.900 and 0.836, respectively. Conclusion:The level of urinary NGAL can effectively predict the AKI in the premature infants.

Objective:To observe the influence of preemptive analgesia by nalbuphine combined with flurbiprofen in the platelet activity indexes of the patients with decompensated cirrhosis, and to explore its quality of analgesia in the patients with decompensated cirrhosis. Methods:A total of 120 patients with decompensated cirrhosis were randomly divided into combination group (40 cases, given nalbuphine combined with flurbiprofen before anesthesia), nalbuphine group (40 cases, given nalbuphine before anesthesia) and flurbiprofen group (40 cases, given flurbiprofen before anesthesia). The postoperative patient-controlled intravenous analgesia (PCIA) was carried out with sufentanil in all the patients. The indexes of visual analog score (VAS), PCIA compression number, sufentanil consumption, Ramsay score,thrombelastogram (TEG) indexes (R value, K value, α angle, MA value and CI value) and the maximum aggregation rate by arachidonic acid (MARAA) and β-endorphin (β-EP) level,recovery time, and agitation occurrence rate of the patients in various groups were recorded 12 h and 24 h after operation. Results:The indexes of VAS, PCIA compression number, sufentanil consumption and β-EP level of the patients in combination group were lower than those in the other groups 12 and 24 h after operation (P<0.05), but the differences of the above indexes of the patients in nalbuphine group and flurbiprofen group were not significant (P>0.05). The Ramsay scores of the patients in three groups had no differences at 12 and 24 h after operation (P>0.05). Compared with nalbuphine group, the R values and K values of the patients in combination group and flurbiprofen group at 12 and 24 h after operation were increased (P<0.05), and the α angle, MA values, CI values and MARAA of the patients in combination group and flurbiprofen group were decreased(P<0.05),but the above indexes of the patients in combination group and flurbiprofen group had no significant differences (P>0.05). The recovery time of the patients among three groups had no significant difference (F=2.054, P=0.102). The agitation occurrence rate during recovery period of the patients in case group was the highest (F=5.624, P=0.001), and there was no significant difference in the agitation occurrence rate during recovery period of the patiens between nalbuphine group and flurbiprofen group (χ²=3.020, P=0.091). Conclusion:Combination of nalbuphine and flurbiprofen in the patients with decompensated cirrhosis before analgesia can improve the quality of postoperative analgesia and can't inhibit the activity of platelet obviously.

Objective:To explore the safeties and efficacies of temporal cortex approach, transsulcus or fissure microsurgery and drilling drainage in the treatment of the patients with supratentorial hypertensive intracerebral hemorrhage (SHICH),and to provide the references for choosing the operation methods for the SHICH patients. Methods:The clinical materials of 118 patients with SHICH were collected and were divided into temporal cortex approach group (cortex group,n=39), transsulcus or fissure microsurgery group (fissure group,n=31) and drilling drainage group (drilling group,n=48) according to their different operation methods. The age, hematoma volumes, Glasgow Coma Scale Score(GCS), time from onset to operation, hematoma clearance rates,improvement rates 24 h after operation, rebleeding rates after operation, intracranial infection rates, survival rates,and good prognosis rates of the patients in three groups were recorded. Results:The hematoma clearance rate, improvement rate, survival rate and good prognosis rate of the patients in fissure group were significantly higher than those in cortex group (P<0.05). There were no statistical differences in the time from onset to operation, rebleeding rates after operation and intracranial infection rates among the patients in three groups (P>0.05). There were no statistically significant differences in the prognosis indexes(hematoma,GCS,survival rate,and good prognosis rate) of the patients between 30-49 mL and 50-69 mL subgroups in fissure group and drilling group. The age of patients in 50-69 mL subgroup in drilling group was significantly higher than that in fissure group(P<0.05). Conclusion:The efficacies and prognosis of SHICH patients treated by transsulcus or fissure microsurgery and drilling drainage are better than those by temporal cortex approach.Drilling drainage is more suitable to the old patients with above moderate hematoma; and transsulcus or fissure microsurgery is more suitable to the patients with massive hematoma.

Objective:To investigate the reasons for the occurrence of postsurgical gastroparesis syndrome (PGS) in the patients with gastric cancer after gastric cancer operation, and to elucidate the related risk factors and treatment measures for the PGS patients,and to provide the basis for improving the quality of life of the patients with PGS. Methods:The clinical data of 1 015 patients underwent radical gastrectomy were retrospectively analyzed and the patients were divided into PGS group(n=39) and non-PGS group(n=976) according to whether the PGS appeared after operation. Single factor analysis was performed according to age, gender, surgical method and anastomosis in the patients in two groups.The risk factors related to PGS were analyzed. Results:There was no difference in the incidence of PGS in the patients ≥ 56 years old and the patients <56 years old (P>0.05); the incidence of PGS in the male patients was not significantly different from that in the female patients(P>0.05); the incidence of PGS in the patients underwemt radical gastrectomy was higher than that in the patients underwent laparoscopic radical gastrectomy (P<0.01); the incidence of PGS in the patients underwent Billroth Ⅰ anastomosis was lower than that in the patients underwent Billroth Ⅱ anastomosis (P<0.01). All the PGS patients were cured after treated with a combination method of drugs and psychology. Conclusion:The occurrence of PGS after radical gastrectomy has nothing to do with the physiological factors of the patients such as gender and age. The incidence of PGS after laparoscopic radical gastrectomy is lower than that after open radical gastrectomy,and the incidence of PGS after Billroth Ⅰ anastomosis is lower than that after Billroth Ⅱ anastomosis.

Objective:To observe the curative effects of artificial femoral head replacement using sternal wire binding and plate fixation in the treatment of the patients with unstable intertrochanteric fracture, and to provide the basis for its treatment. Methods:A total of 52 elderly patients with intertrochanteric fracture treated with artificial femoral head replacement were divided into sternal wire binding group (the patients were given sternal wire binding of greater trochanter,n=28) and plate fixation group (the patients were given plate fixation of greater trochanter,n=24). The operation time, blood loss, the drainage volumes after operation, the time of getting off the bed first, the incidences of early complications, and the Harris scores of hip joint at the discharge time and 1 month, 3 months, 6 months and 12 months after operation of the patients in two groups were recorded. Results:Compared with plate fixation group, the operation time of the patients in sternal wire binding group was decreased (P<0.05), the blood loss of the patients in sternal wire binding group was decreased (P<0.05), but there were no significant differences in the drainage volumes after operation, the time of getting off the bed first, the incidences of complications after operation, and the Harris scores of hip joint at the discharge time and 1 month,3 months, 6 months, and 12 months after operation of the patients between two groups(P>0.05). Conclusion:Compared with plate fixation, sternal wire binding of the greater trochanter in femoral head replacement in the treatment of the patients with unstable intertrochanteric fracture has the advantages of shorter operative time, less blood loss volume,better postoperative recovery of hip joint function,which is suitable for the clinical application.

Objective:To compare the perioperative and recent curative effects of laparoscopic radical resection of low rectal rectal carcinoma by natural orifice specimen extraction surgery(NOSES) with two-step procedure to turn rectum out from the abdominal cavity and traditional one-step procedure to turn rectum out from abdominal cavity. Methods:The clinical data of 63 patients who were treated by laparoscopic radical resection of low rectal rectal carcinoma by NOSES were analyzed retrospectively. A total of 39 patients were treated with the method of two-step procedure to turn rectum out from the abdominal cavity (two-step procedure group)and 24 patients were treated with the traditional one-step procedure to turn rectum out from the abdominal cavity(traditional one-step procedure group). Results:The maximum tumor diameters, the successful rates, tumor fragmentation rates, operation time, intraperative bleeding loss, postoperative exhaust time, anastomotic leakage rates after operation,and the short-term-follow-up results of the patients in two groups were recovered. The specimens of 39 patients in two-step procedure group were successfully pulled out from the anus, the anastomosis was finishehed after resection, and the NOSES of low rectal carcinoma was successfully completed,while 3 cases were failed of 24 cases in traditional one-step procedure group, the successful rate was decreased(P=0.024).After the specimen were pulled out from the anus,the fragmentation rate of the patients in two-step procedure group was lower than that in traditional one-step procedure group (P=0.004). The operation time of the patients in two-step procedure group was less than that in traditional one-step procedure group (t=4.266,P<0.01).There were no significant differences in the maximum diameters of tumor(t=-0.406,P=0.686),intraoperative bleeding loss(t=1.799,P=0.075), postoperative exhaust time (t=-0.144,P=0.886), postoperative anastomotic leakage rate(P=1.000)of the patients between two groups. During 3 to 24 months of follow-up,there were no significant differences in the recurrence rate(P=0.140) and metastatic rate of carcinoma (P=0.36) of the patients between two groups. Conclusion:Two-step procedure to turn rectum out from the abdominal cavity can reduce the pressure on the tumor during turning the rectum out,which can save the operation time and get good short-term curative effect during the radical resection of low rectal carcinoma by NOSES.

Objective:To analyze the clinical diagnosis and treatment and prognosis of the patients with endometrial dedifferentiated carcinoma, and to provide the evidence for its diagnosis and treatment. Methods:The general data, auxiliary examinations,treatment methods and pathological features of 12 patients with endometrial dedifferentiated carcinoma were analyzed retrospectively;literature review was performed,and its diagnosis and treatment methods were analyzed. Results:Among the 12 patients with endometrial dedifferentiated carcinoma, 5 patients in late clinical stage all died, and the average survival time was 10 months;the remaining 7 patients in early clinical stage got a disease-free survival time of 43 months.Twelve patients underwent total hysterectomy, bilateral accessory resection and pelvic lymphadenectomy, of which 2 patients underwent omentectomy and appendectomy, with an average survival time of 8 months; a total of 10 patients remained the omentum majus and appendix, with an average survival time of 24 months.Of the 12 patients, a total of 10 patients received radiotherapy or chemotherapy with an average survival time of 32 months, and 2 patients did not receive the radiotherapy or chemotherapy with an average survival time of 16 months. Conclusion:The patients with endometrial dedifferentiation carcinoma in late clinical stage have shorter survival time and worse prognosis than the patients in early clinical stage.The survival time of patients without chemotherapy or radiotherapy is shorter and the prognosis is worse than those of the patients with chemotherapy or radiotherapy.When the lesion invades omentum majus or appendix,omentectomy and appendectomy are the routine surgical methods for endometrial dedifferentiated carcinoma; resection of the omentum majus or appendix may have no significant effect on the survival time of patient.

Objective:To observe the efficacy of Endobutton combined with the suture in the treatment of Meyers-Mckeever-Zaricnyj type Ⅳ tibial eminence fracture, and to explore the characteristics and treatment methods of Meyers-Mckeever-Zaricnyj type Ⅳ fracture. Methods:The clinical data of seven patients with Meyers-Mckeever-Zaricnyj type Ⅳ tibial eminence fracture were collected;the literature review was performed,and the surgical treatment methods,key points of operation and precautions of the patients were analyzed. Results:After arthroscopic debridement, 3 kinds of main fractures were found:anterior cruciate ligament connected fragment(ACLCF), anterior horn of lateral meniscus connected fragment(ALMCF), and posterior-medial cartilage connected fragment(PMCCF). The PMCCF was first fixed after pre-reduction from back to front and from deep to shallow orders, then the ALMCF and ACLCF were fixed, avoiding the secondary displacement and bone cutting. The follow- up periods of 7 patients were 6-12 months, with an average followed-up period of 7 months. The front drawer test (-), Lachman test (-);the average Lysholm score of patients before operation was less than that in the last follow-up period (P<0.05).The knee joint stability of patients was good, and the knee joint had no movement disorder and infection. Conclusion:The intra-operative reduction order is crucial during the operation of Meyers-Mckeever-Zaricnyj type Ⅳ tibial eminence fracture. Perfect fixation can be achieved by using Endobutton combined with suture.

Objective:To investigate the clinical features,pathological features,treatment methods and prognosis of 5 patients with sarcomatoid carcinoma of throat. Methods:The clinical data of 5 patients with sarcomatoid carcinoma of throat confirmed by pathology were retrospectively analyzed,and the relevant literatures were reviewed. Results:Among 5 patients with sarcomatoid carciroma of throat,4 patients were male and 1 patient was female,with the onset age from 55 to 77 years old. Among them, 3 patients had a long-term smoking history and 2 patients had a long-term drinking history.The chief complaints of the patients were pharyngeal pain,foreign body sensation in the pharynx,hoarseness and dyspnea,etc.All 5 patients underwent electronic laryngoscope examination,and 1 case presented cauliflower mass,while the other patients presented smooth mass.CT examination was performed in 2 patients,and the mass showed soft tissue density shadow without bone destruction. All 5 patients received surgical treatment,including 2 cases of adjuvant radiotherapy after operation and 1 case of chemotherapy after operation.The postoperative histopathological examination and immunohistochemical staining results showed that the diagnosis of 5 patients was sarcomatoid carcinoma.The postoperative follow-up time was 1-36 months,and the follow-up rate was 100%;among 5 patients,1 case survived and 4 cases died. Conclusion:Sarcomatoid carcinoma of throat has no specifically clinical characteristics,and mainly present polypoid growth and is easy to be misdiagnosed.Its malignant degree is higher than that of squamous cell carcinoma,and it is easy to relapse and metastasis in the late stage. Its prognosis is poor,the diagnosis depends on the pathological examination,and surgical resection is preferred.

Objective:To investigate the clinical features of extra-axial medulloblastoma(MB) in the cerebellopontine angle(CPA) area and analyze the imaging features and clinical prognosis, and to provide the reference for its clinical diagnosis and treatment. Methods:The clinical data of two patients diagnosed as extra-axial MB in CPA area were collected;the literature review was performed,and the clinical characteristics,diagnostic and treatment methods were analyzed. Results:Two patients were admitted to the hospital due to headache accompanied by hearing loss.The magnetic resonance imaging (MRI) results indicated a mass in the CPA area.All the two patients were treated with the cerebellopontine angle tumor resection. The operation of the two patients was successful, and the tumor located outside the axis seen in the operation, which was confirmed as MB by postoperative pathology. No recurrence or metastasis of tumor was found 15 months after follow-up. Conclusion:The possibility of MB should be considered before operation for the occupying lesions in the extra-axial CPA area.The active operation can improve the prognosis and prolong the survival of the patients.

Objective:To analyze the clinical features and treatment methods of one infant with low-grade malignant myofibroblastic sarcoma(LGMS) in the left temporal region, and to provide the basis for its diagnosis. Methods:The clinical data of one infant with LGMS in the left temporal region were collected, and the clinical features, diagnosis and treatment of the patient with LGMS were analyzed combined with the relative literatures. Results:A 4-month-old infant was admitted because of left temporal mass for 20 d. The CT image showed a subcutaneous space-occupying lesion in the left temporal region, the CT value was 20-40 HU, and the size was about 2.0 cm×2.0 cm×2.0 cm and the lesion invaded the skull. The enhanced MRI images showed an irregular lesion in the left temporal region, the lesion invaded the skull, the border with the surrounding brain tissue was still clear, the size was about 1.9 cm×1.6 cm×2.0 cm, and the T1WI, T2WI and FLAIR results presented as isointensity in the lesion. The lesion showed inhomogeneous enhancement.The CT and MRI results showed that the subcutaneous tumor invaded the skull, suggesting that there may be malignant tumor. The patient underwent left temporal subcutaneous intracranial and extracranial mass operation to completely resect the lesion, and the pathological diagnosis was LGMS. The patient did not receive radiotherapy or chemotherapy after operation. There was no recurrence after follow-up for 16 months. Conclusion:LGMS is a kind of invasive tumor without specific imaging findings. Total resection is an effective treatment. The pathological diagnosis is the gold standard for the diagnosis of LGMS. The LGMS patients should be given regularly follow-up after operation.

Objective:To study the clinical characteristics of eucine-rich glioma-inactivated1protein(LGI1) antibody positive limbic encephalitis (LE), and to provide the reference for its diagnosis and streatment. Methods:The clinical data of a patient with anti-LGI1 antibody positive LE who presented the hypothalamus symptoms was analyzed, the literature review was performed, and the diagnosis and treatment methods of the LE patients were summarized. Results:The patient showed the following symptoms 2 months before hospitalisation,such as sexual dysfunction,overeating, taciturnity, daytime hyposia, intermittent sweating, fever without inducement, hyponatremia, intermittent memory loss and faciobrachial dystonic seizure (FBDS), and the above symptoms progressively exacerbated. After hospitalization,the Mini-Mental State Examination(MMSE)score was 22;the patient could't finish the evaluation of Montreal Cognitive Assessment Scale(MoCA); the serum anti-LGI1 antibody was positive (1:100),and the cerebrospinal fluid anti-LGI1 antibody was positive(1:10). The MRI results showed T1 low signal, T2, Flair high signal, DWI high signal in bilateral hippocampus, amygdala and medial temporal lobe, especially in the right side. The patient was diagnosed as anti-LGI1 antibody positive LE, whose symptoms relieved rapidly after administration of immunoglobulin and steroid therapy. After 6- month-follow-up, the patient recovered completely. Conclusion:The positive results of serum and cerebrospinal fluid anti-LGI1 antibody in the patients combined with the MRI results of head can confirm the diagnosis of anti-LGI1 antibody positive LE. The hypothalamus symptoms, memory loss and FBDS are improved rapidly after immunoglobulin and steroid therapy. The hypothalamic injury possibly exist in the early-stage of anti-LGI1 antibody positive LE.

Objective:To analyze the diagnosis and treatment methods of one acute gout attack patient with persistent medium and high fever as the first manifestation, and to improve the understanding of the clinicians for acute gout attack with persistent medium and high fever as the first manifestation. Methods:The cliniccal data of an elderly male patient with fever of unknown cause was collected; the literature review was perfomed and the diagnosis and treatment methods of the disease were explored. Results:The patient was admitted to hospital because of persistent fever for 4 d. The count of white blood cells in the routine blood test was high and the left lower lobe inflammation was found in the chest CT scanning. The patient was given the active anti-infection treatment for 10 d,but there was no obvious relief of fever manifestation, and the count of white blood cells remained the high level; no positive results inducing fever were found through comprehensive laboratory examinations and imageological examination. The gout nodules on both feet of the patient were found by physical examination and the level of uric acid was high, so the patient was given experimental anti-gout treatment combined with his previous history of gout. The patient's fever manifestation gradually relieved, the count of white blood cells in routine blood test and the level of uric acid were gradually decreased to the normal levels. The acute gout attack was considered as the cause of fever. Conclusion:For the patients with fever of unknown cause, especially with medium and high fever, if the systematic anti-infection treatment is not effective,the possibility of acute gout attack should be considered.

Objective:To construct a humanized mouse models with human immune system and human tumor growth, and to provide the basis for human tumor immunological research and evaluation on the efficacy of tumor immunotherapeutic durgs. Methods:The humanized mice with human immune system reconstruction were established by intravenous injection of human CD34⁺ fetal liver cells combined with implantation of human fetal thymic tissue under the renal capsule of NOD/SCID IL 2rg-/- after sub-lethal total body irradiation. The peripheral blood was collected,and the immune cell reconstitution was monitored every 3 weeks from the 3th week after humanization, including the ratios of CD45⁺, CD33⁺, CD3⁺, CD3⁺CD4⁺, CD3⁺CD8⁺,and CD56⁺ cells. The human A375 melanoma cells were subcutaneously inoculated at the 15th week after humanization,and the tumor volume was measured every 5 d.. Six weeks after tumor inoculation, the mice were sacrificed and the composition of human immune cells as well as their phenotypes in peripheral blood, bone marrow, lymph node,spleen and tumor tissue were measured by flow cytometry. The composition of human immune cells as well as their phenotypes in tumor tissue were measured by immunohistochemistry. Results:At the 15th week after humanization, the ratio of human T cells was (33.53±8.57)% and the ratio of human B cells was (43.33±10.05)% in the peripheral blood of the humanized mice. The human melanoma grew slowly during the first 3 weeks, and the human tumor sizes expanded rapidly and reached (1 564±334.3) mm³ at the 6th week after tumor inoculation. The immunohistochemical and flow cytometry analysis results showed that a large number of human T cells, which mainly composed by CD8⁺ T subsets, infiltrated in the human melanoma. The ratio of CD8⁺/CD4⁺ in tumor tissue was significantly higher than those in the peripheral blood, bone marrows, lymph nodes and spleen tissue (P<0.01). The levels of PD-1 molecules expressed in tumor-infiltrated human CD4⁺ T and CD8⁺ T cells were higher than those in the peripheral blood and immune organs. Conclusion:A humanized mouse models with human immune system and allogeneic human melanoma is successfully established. The high levels of human immune cells, including human T cells and B cells, can be detected in the peripheral blood, lymph organs, and human tumor tissue in this mouse model.