Objective: To treat the cervical cancer HeLa cells by using calcium inhibitors combined with chemotherapy drugs,and to explore the effect of Ca²⁺ on autophagy in cancer cells,and to provide basis for tumor treatment and drug research.Methods: The human cervical cancer HeLa cells were at logrithmic growth phase randomly divided into control group,cisplatin group,2-APB group,and cisplatin + 2-APB group(combination group).MTT assay was used to measure the survival rates of HeLa cells; Ca²⁺-sensitive fluorescent dye was used to detect the levels of Ca²⁺ in cytoplasm and mitochondrial; the colocalization of LC3 and P62 were examined under confocal microscope.Results: The MTT assay results showed that compared with control group,the survival rates of HeLa cells in cisplatin and combination groups were signifcantly decreased(P<0.05).Compared with cisplatin group,the survival rate of HeLa cells in combination group was significantly increased(P<0.05).The Ca²⁺-sensitive fluorescent dye detection results showed that compared with cisplatin group,the mitochondrial free Ca²⁺ level and the fluorescence intensity of colocalization of LC3 and P62 protein in combination group were significantly decreased.Conclusion: Chemotherapy drugs can induce the autophagy of tumor cells,and calcium inhibitors can increase the sensitity of tumor cells to chemotherapy drugs by reducing the autophagy levels.

Objective: To investigate the effect of a new type of mesoporous bioactive glass with a hierarchical pore structure on the proliferation and differentiation of osteoblasts, and to clarify its osteogenetic differentiation effect. Methods: The MBG structure was observed by scanning electron microscope(SEM);the MG63 cells at logarithmic growth phase were selected and divided into MBG group(cultured with MBG), NBG group(cultured with NBG) and blank control group(cultured with blank cultrue medium), and they were cultured for 1,3,and 5 d.The proliferation rate of MG63 cells was analyzed by MTT method; enzyme-linked immunoassay method was used to analyze the alkaline phosphates(ALP) activity in MG63 cells; real-time PCR was used to detect the relative expression levels of Sp7 mRNA and Runx2 mRNA in MG63 cells after cultured for 24 h. Results: The SEM results showed that the MBG had porous scaffold structure.Compared with blank control group and NBG group,the proliferation rates of MG63 cells in MBG group at 1,3,and 5 d were significantly increased(P<0.01).Compared with blank control group and NBG group, the activities of ALP in the MG63 cells in MBG group at 1,3,5 d were significantly increased(P<0.01). Compared with blank control group and NBG group,the relative expression level of Sp7 mRNA in the MG63 cells in MBG group was significantly increased 24 h after culture(P<0.01), while the relative expression level of Runx2 mRNA in the MG63 cells in MBG group had no significant differences (P>0.05).Conclusion: Compared with NBG,the MBG presents the promotive effect on the proliferation and differentiation of osteoblasts in vitro with low cytotoxicity.

Objective: To express the FLAG-Mu2 protein in drosophila S2 cells,and to investigate its subcellular localization and to purify and identify the FLAG-Mu2 protein complex,and to provide basis for the study of Mu2 protein.Methods: The vector was transfected into drosophila S2 cells. The subcellular localization of FLAG-Mu2 protein was tested by confocal laser scanning microscope. The expression levels of FLAG-Mu2 protein in nuclear extractions were examined using nuclear and cytoplasmic protein extraction method. The FLAG-Mu2 complex was preciptated by precipitation technology and purified by chromatographic steps and the components were analyzed by mass spectrometry.Results: The confocal laser scanning microscope results showed that the FLAG-Mu2 protein mainly expressed in nucleus.The nuclear and cytoplasmic protein extraction results showed that the fusion protein,with a relative molecular weight of 140 000,mainly existed in nuclear extraction. The FLAG-Mu2 complex was purified and analyzed by mass spectrometry,and its results showed that there were 5 components,including pif1A,CG31782,mip120,CG31690,and G protein-sa60A,may be included in FLAG-Mu2 complex.Conclusion: The FLAG-Mu2 protein mainly localizes in the nucleus of drosophila S2 cells; a total of 5 kinds of proteins are found to be potential components of FLAG-Mu2 protein complex.

Objective: To observe the effects of agiophyllum oligosaccharides(AOS) on the general characterization,blood glucose,blood lipid,and insulin sensitivity of the diabetic Goto-Kakizaki(GK) rats,and to disscuse the curative effect of AOS in the patients with type 2 diabetic mellitus.Methods: A total of 60 type 2 diabetic GK rats were randomly divided into model control (MC),glenn benzene urea(GLB),metformin (ME),low (AOS-L),medium(AOS-M),and high doses (AOS-H) of AOS groups,and the homologous Wistar rats were selected as normal control(NC) group;there were 10 rats in each group.All animals were administered with AOS by gavage for 8 weeks.The body weights,food intakes and water intakes of the rats were tested each week.The level of blood glucose was tested by blood glucose meter test for every 2 weeks.The oral glucose tolerance and blood hematoglobin ALC(HbAlc) levels were analyzed after 8 weeks and the serum levels of fasting serum glucose(FPG),fasting plasma insulin (FINS),glycosylated serum protein(GSP), triglyceride (TG),total cholesterol (TC),high-density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) were analyzed in the end of test,at the same time the insulin sensitivity index(IAI) was calculated.Results: Compared with MC group,the general state of rats in administration groups were improved significantly,and the food and water intakes of the GK rats in administration groups were reduced in a certain degree.The food intakes of rats in AOS-M group and AOS-H group were closed to those in GLB group and ME group,and the decline degrees of water intake of rats in different doses of AOS groups were less than those in ME group and GLB group.Compared with MC group,the levels of random blood glucose,HbAlc, and GSP of the rats in different doses of AOS groups(P < 0.05 or P < 0.01) were significantly,especially in AOS-M and AOS-H groups,while there were no significant differences in the levels of random blood glucose,HbAlc, and GSP of rats between different doses of AOS groups(P>0.05).Compared with MC group,the oral glucose tolerance levels of rats in all administration groups were increased and the glucose tolerance areas under curve(AUC) were decreased(P < 0.05 or P < 0.01),especially in AOS-M group.The FPG levels of rats in AOS-M, GLB, and ME groups were obviously lower than that in MC group (P<0.05).The IAI of rats in administration groups were higher than that in MC group(P<0.05).Compared with MC group,the serum levels of TG,TC,and HDL-C of rats in AOS-L group were obviously decreased(P < 0.05 or P < 0.01),and the serum levels of TG and HDL-C in AOS-M group were decreased(P<0.05).Conclusion: AOS can improve the general characterization of GK rats,and reduce the blood glucose level,regulate the blood lipid level and increase the insulin sensitivity.The medium dose of AOS has the better effect.

Objective: To study the promotive effects of leptin and human placenta-derived mesenchymal stem cells(HPMSCs) on healing of combined radiation-wound injury of backs in the rats, and to clarify the synergistic effects of HPMSCs and leptin on healing of combined radiation-wound injury.Methods: A total of 30 Wistar rats were randomly divided into HPMSCs transfected with leptin group (n=10), HPMSCs group (n=10), and blank control group (n=10). The model of combined radiation-wound injury was constructed.The HPMSCs transfected with leptin,the HPMSCs transfected with empty vector, and normal saline were transplanted subcutaneously into the rats in various groups 24 h after radiotherapy, respectively. The color, dermatoglyph,hardness,temperature,haigrowth,exudation,necrotic aera coverage and acupuncture bleeding reaction of the rats in various groups were observed. The survival rates of flap of the rats in various groups was detected.The histomorphology of flap after radiotheraphy in various groups was observated by HE staining. The expressions of von Willebrand factor(vWF) and leptin in flap of the rats in various groups after radiotheraphy were detected by immunohistochemical method.Results: On the 7th day after administration, the boundary of survival area and necrotic area of the rats in various groups were clear.The nectotic area of flap was hard and the color was ruddy,and the nectotic area of flap of the rats in HPMSCs transfected with leptin group and HPMSCs group were less than that in blank control group.On the 7th day after administration, the survival rates of flap of the rats in HPMSCs transfected with leptin group and HPMSCs group were higher than that in blank control group(P<0.05).The survival rate of flap of the rats in HPMSCs transfected with leptin group was higher than that in blank control group(P<0.05). The HE staining results showed that on the 14th day after administration, there were the most abundant granulation tissues, new capillaries and fibroblasts and there were less inflammatory cell infiltration around the capillaries; the collagen arranged irregularly in HPMSCs transfected with leptin group.On the 14th day after administration, the immunohistochemical staining results showed that there were a lot of yellow particals (positive expression) in HPMSCs transfected with leptin group and HPMSCs group.A lot of leptin was shown in HPMSCs transfected with leptin group.Conclusion: In the role of local microenvironment, HPMSCs can differentiate into endothelial cells and promote the formation of new capillaries.HPMSCs and leptin have synergistic effects in the process of wound healing, and the HPMSCs with gene-modified or not could significantly increase the survival rate of flap after radiotherapy, and the HPMSCs transfected with leptin have better effect.

Objective: To study the feasibility of hepatitis C virus (HCV)multi-epitope DNA vaccines prepared by minimalistic immunologically defined gene expression,and to clarify its potential application value in the field of DNA vaccines.Methods: A minimalistic immunologically defined gene expression DNA vaccine(1 346 bp) containing CMV promoter,HCV 1b subtype multi-epitope, and bovine growth hormone (BCG) polyA sequence was prepared by artificial synthesis and PCR methods,then it was named M-HCV-epi.A control DNA fragment,which structure gene was replaced by a DNA fragment without homologous to HCV,was prepared and named V-pcDNA3.1. A total of 12 ICQ mice were randomly divided into experimental group(n=6) and control group(n=6);the mice were subcutaneously injected with M-HCV-epi and V-pcDNA3.1 with the dosage of 20 μg,respectively.The levels of interferon-γ (IFN-γ) mRNA in spleen cells of the immunized mice were detected by QRT-PCR method. A total of 3 HCV 1b subtype epitope polypeptides and a control polypeptide were synthetically prepared.The levels of IFN-γ in culture supernatant of spleen cells were detected by ELISA method after the cells were stimulated with the polypeptides prepared above.Results: Compared with control group,the level of IFN-γ mRNA in spleen cells of the mice in experiment group was increased to (1.50±0.18) times(P<0.05); compared with control group,the level of IFN-γ in the culture supernatant of spleen cells of the mice in experiment group was increased (P<0.05).Conclusion: The HCV multi-epitope DNA vaccine with minimalistic immunologically defined gene expression can induce the cellular immune response,and may have application values in the field of DNA vaccines.

Objective: To explore the effect of SOX2 on the neural differentiation of dental pulp stem cells (DPSCs), and to provide basis for studying the mechanism of neural differentiation of DPSCs.Methods: The DPSCs (DPSCs group) were isolated from human dental pulp of third molar, and the SOX2 over exression DPSCs (DPSCs-SOX2 group) were established via retrovirus infection. The empty vector-infected DPSCs (DPSCs-vector group) were regarded as control group. The DPSCs in various groups were induced into neural progenitor cells (NPCs) by neural differentiation medium. The proliferation index (PI) of NPCs in various groups was evaluated by CCK-8 method; the expression levels of Nestin and Pax6 gene in NPCs in various groups were detected by qPCR and FACS; the differentiation abilities of NPCs and the expression efficiencies of β3-Tubulin after differentiation in various groups were evaluated by immunofluorescence and FACS methods.Results: All of normal DPSCs, DPSCs-vector and DPSCs-SOX2 derived from NPCs could be differentiated into NPCs successfully through the induction of neural differentiation medium. Compared with DPSCs and DPSCs-vector groups, the PI of DPSCs in DPSCs-SOX2 group was increased (P<0.05), and the expression levels of Nestin and Pax6 in NPCs were increased (P<0.05). The expression efficiency of β3-Tubulin in differentiated NPCs in DPSCs-SOX2 group was higher than those in DPSCs and DPSCs-vector groups (P<0.05).Conclusion: SOX2 could have a certain promotive effect on the neural differentiation of DPSCs.

Objective: To discuss the hypoglycemic effects of saponins, flavonoids, and polysaccharides of fenugreek with different component compatibilities on the type 1 diabetic rats,and to study the optimal compatibility proportion of three kinds of components in reducing the blood glucose level.Methods: The saponins, flavonoids,and polysaccharides were extracted from the fenugreek and then they were separated and purified by column chromatography and decoction and alcohol sedimentation technique, respectively. The compatibility proportion of polysaccharides, flavonoids and saponins as well as the grouping were determined by uniform design method and the ratioes of three kinds of components to fenugreek. The administration groups were divided into low dose of polysaccharides compatibility group (polysaccharides:saponins:flavonoids 3:6:1), middle dose of polysaccharides compatibility group (polysaccharides:saponins:flavonoids 12:8:1),and high dose of polysaccharides compatibility group (polysaccharides:saponins:flavonoids 10:4:1). A total of 35 type 1 diabetic rats induced by streptozotocin(STZ) were divided into model control group (n=9), positive drug group (n=7) and low (n=6), middle(n=6), and high(n=7) doses of polysaccharides compatibility groups, respectively. Normal control group (n=11) was set up at the same time. The rats in normal control group and model control group were given pure water with the dose of 10 mL·kg^-1·d^-1. The metformin hydrochloride was administrated to the rats in positive drug group wtih the dose of 0.018 g·kg^-1·d^-1. The components of fenugreek were given to the rats in three doses of polysaccharides compatibility groups by the proportion mentioned above by intragastric administration for 28 d with the dose of 0.45 g·kg^-1·d^-1. The levels of fasting blood glucose(FBG), urinary protein in urine,serum triglyceride (TG), total cholesterol(TC), and advanced glycation endproducts(AGEs) in the rats were determined. The morphology of pancreas and kidney tissues of the rats were observed by HE staining.Results: The levels of FBG, urinary protein,serum TG, TC, and AGEs of the rats in model group were increased significantly (P<0.01) compared with normal control group. The levels of FBG of the rats in three doses of polysaccharides compatibility groups and positive drug group were decreased gradually and were decreased significantly on the 28th day after administration (P < 0.05 or P < 0.01) compared with model group; the levels of urinary protein,serum TG, TC, and AGEs were significantly decreased (P<0.01). The levels of urinary protein, TG, TC, and AGEs of the rats in low dose of polysaccharides compatibility group were decreased most significantly. The damage of pancreas and kidney tissues of rats in three doses of polysaccharides compatibility groups and positive drug group were improved compared with model group. Conclusion: The three components of fenugreek (polysaccharides:saponins:flavonoids) at the compatibility proportion of 3:6:1 can better regulate the blood glucose of type 1 diabetic rats and protect the pancreas and kidney tissues of rats.

Objective: To investigate the induction of transforming growth factor beta 1 (TGF-β1) on epithelial to mesenchymal transition (EMT) in glioma U87 cells, and to provide evidence to clarify the mechanism of glioma invasion growth.Methods: The glioma U87 cells were treated with different concentrations of TGF-β1 in vitro and divided into 2.5×10^-6,5.0×10^-6,10.0×10^-6,and 20.0×10^-6g·L^-1 TGF-β1 groups,and 0×10^-6g·L^-1 TGF-β1 treatment was regarded as blank control group.The relative expression levels of epithelial marker(E-cadherin),mesenchymal markers(N-cadherin,Vimentin),and transcription factors(Snail,Slug, and Zeb1 proein) in the U87 cells in various groups were detected by Western blotting method; the morphology of U87 cells was observed by inverted microscope; the relative wound space of U87 cells was examined by scratch healing assay.Results: With the increasing of concentrations of TGF-β1,the relative expression levels of E-cadherin in glioma U87 cells in 2.5×10^-6,5.0×10^-6,10.0×10^-6, and 20.0×10^-6 g·L^-1 groups were gradually decreased compared with blank control group (P < 0.05 or P < 0.01).The relative expression levels of N-cadherin and Vimentin showed an increasing trend,and the relative expression levels of N-cadherin were increased by 11.5%(P>0.05),39.4%(P>0.05),51.0%(P<0.05), and 76.9%(P<0.05) in 2.5×10^-6,5.0×10^-6,10.0×10^-6, and 20.0×10^-6 g·L^-1 TGF-β1 groups compared with blank control groups. The relative expression levels of Vimentin in the U87 cells in TGF-β1 10×10^-6 and 20×10^-6g·L^-1 groups were higher than those in blank control group(P<0.01). The relative expression levels of Snail,Slug and Zeb1 proteins in the U87 cells in 5.0×10^-6,10.0×10^-6, and 20.0×10^-6g·L^-1 groups were higher than those in blank control group(P < 0.05 or P < 0.01). Compared with blank control group,the morphology of U87 cells in 10×10^-6g·L^-1TGF-β1 group became more stretching,long and narrow, and the cell migration ability was also significantly enhanced.Conclusion: Within a certain concentration range,TGF-β1 could induce EMT in glioma U87 cells in a concentration-dependent manner.

Objective: To detect the inhibitory effect of gallic acid on the proliferation of human non-small cell lung cancer A549 cells,and to investigate the molecular mechanisms of its drug toxicity.Methods: The human non-small cell lung cancer A549 cells were cultured in vitro.The cells were divided into control group,low,middle,and high dosages of gallic acid groups (0,300,500 and 750 μmol·L^-1).The survival rates of cells were tested by MTT method; the morphology of A549 cells were tested by Switzerland and Giemsa staining; the apoptotic rates of A549 cells and the levels of reactive oxygen species (ROS) in A549 cells were analyzed by FCM; the expression levels of Fas and FasL in the A549 cells in various groups were detected by Western blotting method; the activities of catalase(CAT),superoxide dismutase (SOD)and glutathione peroxidase(GSH-Px) in the A549 cells in various groups were analyzed by spectrophotometry.Results: Compared with gallic acid treatment for 6 h,the survival rates of A549 cells treated for 12 and 24 h in different dosages of gallic acid groups were significantly decreased(P<0.05),and the cell survival rates were gradually decreased with the prolongation of treatment time.Compared with control group,the survival rates of A549 cells in different dosages of gallic acid groups were significantly decreased after treatment for 24 h (P<0.05).The morphology of A549 cells in different dosages of gallic acid groups showed the typical apoptosis which were observed by Switzerland and Giemsa staining,such as the cytoplasmic shrinkage, small and round cell volume and so on.The results of and so on apoptotic examination showed that the apoptotic rates were increased with the increasing of dosage of gallic acid.The expression levels of Fas and FasL in A549 cells were increased with the increasing of gallic acid dosages.The expression levels of Fas were positively correlated with the expression levels of FasL (r=0.92,P<0.05).Compared with control group,the levels of ROS in A549 cells in low,middle and high dosages of gallic acid groups were increased (P<0.05).Compared with control group,the activities of CAT in A549 cells in different dosages of gallic acid groups were also increased,but there were no significant differences (P>0.05); the activities of CAT in A549 cells in different dosages of gallic acid groups were reduced with the increasing of gallic acid dosage.The activity of SOD in low dosage of gallic acid group was higher than that in control group (P<0.05).The activities of SOD in middle and high dosage of gallic acid groups were higher than that in control group (P<0.05),but there were no significant differences between various groups (P>0.05).The activities of SOD were reduced with the increasing of gallic acid dosages.The activity of GSH-Px in A549 cells in low dosage of gallic acid group was higher than that in control group,but there was no significant difference between various groups (P>0.05).The activities of GSH-Px in middle and high dosages of gallic acid groups were lower than that in control group,but there were also no significant differences between various groups (P>0.05).The activities of GSH-Px were also reduced with the increasing of gallic acid dosage.There were no significant differences of the activities of CAT,SOD, and GSH-Px in A549 cells between low,middle, and high dosages of gallic acid groups(P>0.05).Conclusion: Gallic acid can inhibit the proliferation of lung cancer cells by oxidative damages,and the Fas/FasL signal pathway might be one of the mechanisms for apoptosis-induced by gallic acid.

Objective: To detect the expression levels of microRNA-34a(miR-34a) in gingival squamous cell carcinoma (GSCC) tissue and adjacent tissue,and to clarify its correlation with the pathological characteristics of GSCC patients,including tumor size, malignancy degree,age,gender,tumor site and lymph node metastasis.Methods: The tissue samples including 30 GSCC tissues and matched carcinoma adjacent tissues were selected. The expression levels of miR-34a in GSCC tissue and carcinoma adjacent tissue were detected by quantitative reverse transcription-PCR(qRT-PCR) method. The relationship between its expresion level and the clinicopathological characteristics of GSCC patients were analyzed.Results: Compared with carcinoma adjacent tissue,the expression level of miR-34a in GSCC tissue was significantly decreased (t=3.03,P=0.004).The expression level of miR-34a in GSCC tissue had no relationship with the age,gender, and lymph node metastasis of the patients(P>0.05),but had relationship with the tumor size and tumor malignancy degree (P<0.05).Conclusion: The expression level of miR-34a is down-regulated significantly in GSCC tissue.The detection of the expression level of miR-34a contributes to judge the histological subtype.

Objective: To explore the effect of Ginseng and Coptis composition on the biochemical indexes and morphology of kidney tissue in the diabetes nephrophthy (DN) mice,and to provide basis for its application in DN treatment.Methods: Ten C57 mice were selected as normal control group.A total of 240 C57 mice were induced to establish DN models by feeding with high-sugar and high-fat diet combined with small dose of streptozotocin(STZ).The DN mice were randomly divided into model control group,low(80 mg·kg^-1+20 mg·kg^-1),medium(160 mg·kg^-1+40 mg·kg^-1), and high(320 mg·kg^-1+80 mg·kg^-1) doages of Ginseng and Coptis composition groups, total alkaloid from Coptis group(320 mg·kg^-1), total Ginsenoside group(80 mg·kg^-1),and captopril group(8 mg·kg^-1); there were 10 mice in each group. After treated for 8 weeks,the physiological status of C57 mice in various groups were observed.The levels of fasting blood glucose(FBG),microalbuminuria(MAU),serum creatinine (Scr),blood urea nitrogen (BUN),total cholesterol (TC),and triglyceride(TG) of mice in various groups were measured.The pathomorphology of kidney tissue of mice was observed by HE staining.Results: Compared with model control group,the physiological status of C57 mice in different dosages of Ginseng and Coptis composition groups,such as the urine volume and the body weight increase were significantly improved,and the levels of FBG were decreased (P<0.05); the levels of MAU,BUN,TC, and TG were also decreased (P<0.05).The glomerular and renal interstitial lesions of the kidney tissue in different dosages of Giseng and Coptis composition groups were significantly improved.Conclusion: Ginseng and Coptis composition has therapeutical effect on early DN.

Objective: To prepare the hepatocellular injury-conditioned medium (CM), and to simulate the microenvironment of injured tissue in vitro, and to explore the effect of CM on hepatic differentiation of bone marrow mesenchymal stem cells (BMMSCs) in the rats.Methods: The BMMSCs from 1-week-old rats were isolated and cultured by differential adherence and differential gradient method combined with optimized gradient centrifugation and culture time for adherent fluid exchange.The rats aged 6-8 weeks were injected intraperitoneally with 60% carbon tetrachloride solution (0.75 mL·kg^-1) to prepare the hepatocyte injured tissue homogenate supernatant (CM1) and hepatocyte injured serum (CM2). The third passage (P3) BMMSCs were randomly divided into 0%CM group (control group, cultured with 10%FBS L-DMEM), 10%CM1 group, 20%CM2 group, and 10%CM1+10%CM2 group. The morphology of cells was observed under inverted microscope;the expressions of cell-surface marker CD105 in BMMSCs and the positive expression rates of albumin (ALB) in cells in various groups were detected by immunohistochemistry method;the positive rate of lipid droplet formation in the pocess of adipogenic differentiation was detected by oil red O staining; the expression levels of alpha-fetoprotein (AFP) and ALB in the cells in various groups were detected by RT-PCR method;the positive rate of glycogen synthesis was detected by periodic acid-shiff staining; the hepatic differentiation ability of BMMSCs in various groups were identified and compared. Results: After primary culture of bone marrow cells for 72 h, the adherent cells showed round or spindle-shaped, covered with the bottom of the fibrous shuttle,and showed uniform and orderly shape, and the P3 BMMSCs surface markers CD105 showed positive expression.The P3 BMMSCs were induced f or 4 d by dexamethasone, insulin, 3-isobutyl-1-methylxanthine and indomethacin (DX+IBMX+IS+ID), the formation of lipid droplets was observed under microscope. After P3 BMMSCs were induced by CM, the expressions of AFP and ALB were positive,and the staining of cell glycogen was positive. After induced for 7 d, the cytoplasmic contents of P3 BMMSCs were abundant and the cells were round or triangular or liver board-like arrangement in CM groups. After induced for 7, 14 and 21 d, compared with control group, the positive expression rates of ALB in BMMSCs in CM groups were significantly increased (P<0.01); the positive expression rates of ALB in BMMSCs in 10% CM1+10% CM2 group were higher than those in 10%CM1 group and 20%CM2 group (P<0.05). After induced for 7 and 14 d by CM, compared with control group, the positive rates of glycogen synthesis in BMMSCs in CM groups were significantly increased (P<0.01);the positive rates of glycogen in 10%CM1+10%CM2 group were higher than those in 10% CM1 group and 20% CM2 group (P<0.05).Conclusion: CM1 and CM2 could induce the differentiation of BMMSCs into hepatocyte-like cells,and the lower concentrations of CM1 and CM2 can achieve the same effect.The microenvironment provided by CM1 and CM2 is more beneficial to the differentiation of BMMSCs into hepatocytes, which could induce cell differentiation more efficiently.

Objective: To evaluate the role of low expression of let-7 in the prognosis of lung cancer and the clinical significance of relationship between the HMGA2 expression and lung cancer by Meta-analysis method.Methods: The studies about the correlation between the low expression of let-7 and the prognosis of lung cancer and the correlation between the HMGA2 expression and the occurrence and development of lung cancer were collected by searching PubMed,Web of Science,Medline,Cochrane Library,Google, Scholar,CNKI,CBM,and Wanfang,and VIP databases.The StataSE 12.0 software was used for this Meta-analysis.Results: A total of 10 studies about the relationship between the low expression of let-7 and the prognosis of lung cancer (the patients with non-small cell lung cancer) were enrolled in this Meta-analysis.The low expression of let-7 indicated a poor overall survival rate,and the combined value of hazard ratio(HR) was 1.55 (95%CI:1.16-2.09,P<0.05).A total of 9 case-control studies about the relatlonship between the expression of HMGA2 and the occurrence and development of non-small cell lung cancer were selected for this Meta-analysis. The positive expression rate of HMGA2 in lung cancer tissue was significantly higher than that in paracancer normal tissue(OR=5.92,95% CI:3.96-8.85,P=0.000).The expression of HMAG2 in lung cancer tissue was pasitively related with the TNM stage and the lymph node metastasis of the patients(OR=0.26,95%CI:0.16-0.41,P<0.05;OR=0.29,95%CI:0.12-0.71,P<0.05).Conclusion: The low expression of let-7 is closely related to the poor prognosis of lung cancer patients.The highly positive expression rate of HMGA2 in lung cancer tissue is positively correlated with the occurrence and invasiveness of lung cancer.

Objective: To evaluate the effectiveness of 9 kinds of traditional Chinese herb injections combined with transcatheter arterial chemoembolization (TACE) in the treatment of hepatocellular carcinoma and improvement of life quality of the patients using network Meta-analysis.Methods: The Cochrane Library, PubMed, EMbase and CBM, VIP Chinese Journal Database, Wanfang database, and CNKI database were searched by computer. The retrieval time for each database was from establishment time to December 30, 2015. The literature were screened according to the pre-established inclusion and exclusion criteria, and the qualities of these included studies were evaluated. R software was used for statistical analysis and the results of randomized controlled trial (RCTs) were combined and sorted by network Meta-analysis.Results: A total of 86 RCTs including 5 791 patients were finally included. Based on the probabilistic ranking results, the total effective rates were arranged in the descending order, Cantharidin Injection (OR=5.60, 95%CI:3.55-8.84, P=0.920), Kanglaite Injection (OR=5.66, 95%CI:2.96-10.82, P=0.906), Elemene injection (OR=3.34, 95%CI:1.77-6.32, P=0.601), Compound Kushen Injection (OR=2.86, 95%CI:1.84-4.46, P=0.478), Brucea Javanica Oil Injection (OR=2.85, 95%CI:1.94-4.19, P=0.475), Cinobufacini Injection(OR=2.79, 95%CI:2.02-3.85, P=0.452), Delisheng Injection (OR=2.74, 95%CI:1.22-6.15, P=0.448), Aidi Injection (OR=2.71, 95%CI:2.12-3.47, P=0.418) and Kangai Injection (OR=2.37, 95%CI:1.55-3.66, P=0.301). The probabilities of improvement of life quality were ranked from high to low order, Aidi Injection (OR=3.06, 95%CI:1.25-7.47, P=0.669), Kangai Injection (OR=3.38, 95%CI:0.50-22.74, P=0.655), Kanglaite Injection (OR=2.09, 95%CI:0.21-21.38, P=0.599), Delisheng Injection (OR=3.33,95%CI:0.05-210.09, P=0.586) and Cinobufagin Injection (OR=2.39,95%CI:0.47-12.26, P=0.556), Brucea Javanica Oil Injection (OR=2.25,95%CI:0.74-6.81, P=0.547), Cantharidin Injection (OR=1.82,95%CI:0.62-5.35, P=0.464), Elemene Injection (OR=2.20,95%CI:0.08-62.31, P=0.403) and Compound Kushen Injection (OR=1.26,95%CI:0.37-4.27, P=0.325).Conclusion: Cantharidin Injection, Kanglaite Injection, and Elemene Injection are more relatively effective than the other six kinds of traditional Chinese herb injections. Aidi Injection, Kangai Injection, and Kanglaite Injection can improve the life quality better than the other six kinds of traditional Chinese herb injections.

Objective: To investigate the association between PLA2G12A polymorphism and autism spectrum disorder (ASD) in North Han Chinesechildren,and to explore whether PLA2G12A is a susceptibility gene for ASD.Methods: A total of 85 ASD children (case group) and 183 healthy children(control group) were enrolled in our case-control study.Four SNPs (rs6533451,rs2285714,rs2285713,and rs11728699) on PLA2G12A gene were selected and the genotype and allele were tested using Sequenom Mass ARRAY System.The SNPStats program was used for analyzing the Hardy-Weinberg equilibrium test;the differences in genotypic and allelic frequencies of the subjects between two groups were detected; the association between ASD and 4 SNPs was analyzed by genetic models; the Haploview 4.2 software was used to analyze the association between ASD and haplotypes.Results: The distribution of genotypes and alleles of all SNPs sites did not deviate from Hardy-Weinberg equilibrium in both case and control groups (P>0.05). There were no significant differences of the genotypic and allelic frequencies of four SNPs sites of the subjects between case and control groups (P>0.05). There were no significant association between ASD and the gene polymorphisms of four SNPs sites under five genetic models (P>0.05).The haplotypes composed by rs251684,rs2307279,and rs156631 sites were not significantly associated with ASD (P>0.05).Conclusion: PLA2G12A may not be a candidate gene for ASD in North Han Chinese children.

Objective: To discuss the expressions of hMOF protein and mRMA in endometrial cancer tissue,and to illuminate the possible mechanism of hMOF in the occurrence and development of endometrial cancer.Methods: The PT-PCR and immunohistochemistry methods were used to detect the expressions of hMOF protein and mRNA in 38 cases of endomerial cancer tissues and 20 cases of normal endometrial tissues,and to analyze the relationship between its expression level and the clinical characteristics and prognosis of the patients with endometrial cancer.Results: The immunohistochemistry results showed that the positive expression rate of hMOF protein in endometrial cancer tissue was lower than those in normal endometrial tissue(P=0.037); the positive expression rate of hMOF protein in stage Ⅰ patients was higher than those in stage Ⅱ and Ⅲ-Ⅳ patients(χ²=6.591,P=0.037);the positive exprssion rate of hMOF protein in the patients with poor differentiation was lower than those in the patients with moderate and high differentiation(χ²=7.351,P=0.025).The RT-PCR results showed that the percentage of the patients with expression up-regulation of hMOF mRNA in endometrial carcinoma tissue was remarkly decreased compared with normal endometrial tissue(P=0.003).Conclusion: hMOF protein may participate in the occurrence and development of endometrial cancer,and hMOF protein could be a biomarker for prognosis of endometrial cancer.

Objective: To understand the typing of Rh system phenotypes and analyze the positive rate of irregular antibody in Rh system,and to provide basis for blood transfusion of the elderly patients in clinic.Methods: A total of 587 elderly blood tranfusion candidates were included. Microcolumn gel test was applied to detect the antigens of Rh system and identify the irregular antibody in Rh system.Results: There were 7 kinds of Rh blood group phenotypes with postive expression of RhD in 587 patients; the positive rates of irregular antibodies from high to low were CCDEE 38.7%(227/587),CcDEe 34.8%(204/587),CcDee 12.4%(73/587),ccDEE 8.7%(51/587),ccDEe 4.6%(27/587) and CCDEe 0.7%(4/587),ccDee 0.2%(1/587).There were 6 kinds of irregylar antibodies in 587 patients;the positive rate was 1.02%,including 4 cases of Anti-E (66.67%),1 case of Anti-c(16.67%),1 case of anti-M(16.67%).The postive rate of irregular antibody in the patients with blood transfution history and pregnancy history (2.65%,51/89) was higher than those without blood transfusion history or pregnancy history 0.25%,13/98)(χ²=5.11,P<0.05). There were no significant differences of the positive rates of irregular antibody between different nationalities(χ²=0.10,P>0.05).Conclusion: The irregular antibody of Rh system is the major antibody in the eldery blood transtusion candidates.The detection of typing of Rh system phenotypes and identification of irregular antibodies in the patients with blood transfusion history and pregnancy history should be performed to ensure the safty of blood transfusion.

Objective: To explore the risk factors of ovarian hyperstimulation syndrome(OHSS), and to study the intravenously injected with preventive effect of hydroxyethyl starch solution on OHSS.Methods: A randomly prospective clinical trial including 192 fertilization-embryotransfer(IVF-ET) patients with high risk of OHSS was performed in vitro,and the patients were randomly divided into experimental group(n=101) and control group(n=91).The patients in experimental group were intravenously injected with 500 mL hydroxyethyl starch solution per day during 24 h of oocyte retrieval and lasted for 3 d;the patients in control group were not treated with any drugs.The general data,the incidence of OHSS,and the incidence of moderate and severe OHSS of the patients in two groups were compared; the relative factors of moderate and severe OHSS were analyzed by Logistic regression analysis.Results: The Logistic regression analysis results showed that the smaller age and body mass index were the protective factors of OHSS (OR<1)and the higher E2 value on the human chorionic gonadotropin(HCG) injection day and the larger number of follicles taken from the oocytes(OR>1) were the risk factors of OHSS.The incidence of OHSS of the patients in experimental group was 31.68%(32/101),which included 21 cases of moderate OHSS (20.79%) and 11 cases of severe OHSS(10.89%);the incidence of OHSS in control group was 46.15%(42/91),which included 24 cases of moderate OHSS(26.37%) and 18 cases of severe OHSS(19.78%);the incidence of OHSS and severe OHSS of the patients in experimental group was significantly lower than those in control group(χ²=4.23,P<0.05;χ²=2.95,P<0.05).Conclusion: Age,body mass index,E2 value on the HCG injection day,and the number of follicles taken from the oocytes are the related factors of OHSS,and play an important role in predicting the incidence of OHSS;hydroxyethyl starch solution can markedly decrease the incidence of OHSS,and has better preventive effect on severe OHSS.

Objective: To discuss the therapeutic effect of Compound Zikui Yuanyang Capsule in the treatment of the patients with benign prostatic hyperplasia (BPH), and to illuminate the clinical effect of Compound Zikui Yuanyang Capsule in the treatment of the BPH patients with obvious kindney deficency syndrome.Methods: A total of 66 patients were divided into experimental group (given Zikui Yuanyang Capsule) and control group (given placebo). The International Prostate Symptom Score (I-PSS), Qmax,Quality of Life Score (QOL), International Index of Erectile Function(ⅡEF),and testosterone level of the patients in two groups were detected before and after treatment.Results: In Chinese medicine symptoms, the percentages of patients with clinical control,marked effectivness,and effectiveness in experimental group and control group were 10%, 13%, 57% and 0%, 6.25%, 16.00%, respectively, and the total effective rates were 86.00% and 22.25%, respectively; the total effective rate of the patients in experimental group was superior to that in control group (P<0.05). In the main symptoms, the number of the patients with frequent urination disorder, more night urination number, and dysuria in experimental group was less than that in control group (P<0.05). Conclusion: Compound Zikui Yuanyang Capsule has better clinical efficacy and safety in the treatment of the BPH patients with kidney deficiency symptoms.

Objective: To construct a database based on the analysis of the soft tissue profile of Northern Chinese young adults by means of angular measurement made on the standardized photographic records taken under natural head position,and to provide basis for the orthodontists and oral surgeons to make diagnosis and treatment in clinic.Methods: A total of 113 students (67 female and 46 male) who met the inclusion criteria from Jilin University were selected.The right profile records under natural head position were taken.The facial profile of soft tissue was analyzed by coreldraw software.A total of 12 angular parameters were detected.The reproducibility of the measurement was analyzed by Dahlberg's formula.An independent sample t-test was used to analyze the differences in the measurement values of different soft tissue profile angles between different genders.Results: Except for nasolabial angle (Cm-Sn-Ls),mentolabial angle (Li-Sm-Pg), lower facial height angle (Sn-Trg-Me),the method errors of the other measurements were small.The mean values of nasofrontal angle (G-N-Prn),nasal angle (N-Prn/Sn-Cm),mentolabial angle (Li-Sm-Pg),angle of facial convexity (G-Sn-Pg),and angle of total facial convexity (G-Prn-Pg) of the female were greater than those of the male (P < 0.05 or P < 0.01),while the mean values of vertical nasal angle (N-Prn/TVL) and superior triangle (Pg-G-Prn) of the male were greater than those of the female (P<0.01).The standard deviation of nasolabial angle (Cm-Sn-Ls) and mentolabial angle (Li-Sm-Pg) were greater.Conclusion: A database based on the angular analysis of the soft tissue profile of Northern Chinese young adults under nature head position is obtained,which can provide basis for the clinical diagnosis and treatment of the orthodontists and oral surgeons.

Objective: To observe the changes of expression levels of serum soluble Tim-3(sTim-3) and its regulation effect on Th1/Th2 differentiaton in the patients with unexplained recurrent spontaneous abortion(URSA),and to investigate the possible immunmechanism of sTim-3 in the pathological process of URSA. Methods: A total of 52 patients with URSA were regarded as URSA group,and 20 normal pregnancy women at the same period were selected as normal pregnancy group and 20 non-pregnancy women in gestation age were selected as healthy control group.ELISA method was used to detect the levels of serum sTim-3 and galactose binding protein 9(Galectin-9) and the levels of serum Th1 cytokines interferonI-γ(IFN-γ) and IL-2 and Th2 cytokines IL-4 and IL-10 of the subjects in three groups.Results: Compared with healthy control group,the levels of serum sTim-3(t=7.34,P<0.05)and Galectin-9 (t=4.86,P<0.05) of the patients in URSA group were increased. The level of serum sTim-3 of the patients in URSA group was significantly higher than that in normal control group(t=5.63,P<0.05),but the level of serum Galectin-9 of the pathents in URSA group was lower than that in normal control group(t=2.76,P<0.05).The levels of serum IFN-γ and IL-2 of the subjects in normal control group(t=2.81,P<0.05;t=2.84,P<0.05) and URSA group(t=3.66,P<0.05;t=2.91,P<0.05) were lower than those in healthy control group.The levels of serum IFN-γ and IL-2 of the patients in URSA group were higher than those in normal control group(t=6.11,P<0.05;t=5.95,P<0.05).The levels of serum IL-4 and IL-10 of all the subjects in normal pregnancy group and URSA group were higher than those in healthy control group (t=2.53,P<0.05;t=2.52,P<0.05;t=2.69,P<0.05;t=2.33,P<0.05); the IL-4 and IL-10 levels of the patients in URSA group were lower than those in normal control group (t=6.48,P<0.05;t=7.76,P<0.05).Conclusion: The increased expression of sTim-3 and the imbalance of Th1/Th2 homeostasis may be one of the key immunomechanisms of inducing recurrent abortion in the URSA patients.

Objective: To investigate the expressions of Ki-67 and cyclooxygenase-2(COX-2) in colorectal carcinoma tissue and their clinical significances,and to provide basis for the diagnosis and treatment of colorectal carcinoma.Methods: A total of 71 colorectal carcinoma patients were selected.The positive expression rates of Ki-67 and COX-2 in colorectal carcinoma tissue (colorectal cancer group,71 cases),para-carcinoma tissue(para-carcinoma group,71 cases),and normal mucosal tissue (normal group,20 cases) were detected by immunohistochemical SP method.The relationship between the expressions of Ki-67 and COX-2 and metastasis,different stages and prognosis of colorectal carcinoma were analyzed.The correlation between the expressions of Ki-67 and COX-2 was analyzed by Spearman analysis method.Results: The psitive expression rates of COX-2 and Ki-67 in colorectal carcinoma tissue were obviously higher than those in adjacent tissue and normal mucosal tissue(χ²=24.581,P<0.05;χ²=11.538,P<0.05).The positive exprssion rates of COX-2 and Ki-67 in carcinoma tissue of the patients with lymph node metastasis were significantly higher than those in the patients without lymph node metastasis(χ²=5.021,P=0.025;χ²=7.289,P=0.007).The positive expression rates of Ki-67 and COX-2 in carcinoma tissue in the patients at Ⅰ+Ⅱ stage were significantly lower than those in the patients at Ⅲ+Ⅳ stage(χ²=8.570,P=0.003;χ²=11.409,P=0.001).The expressions of COX-2 and Ki-67 in colorectal carcinoma tissue of the patients had a positive correlation(r=0.520,P<0.05).The post-operation medium survival time of the patients were shortened with the increasing of the expression levels of Ki-67 and COX-2, and the 1-,3-,and 5-year survival rates of the patients with positive expressions of COX-2 and Ki-67 were decreased gradually.Conclusion: The abnormal expressions of COX-2 and Ki-67 play an important role in the occurrence and development of colorectal carcinoma and may be responsible for the occurrence,development,invasion and metastasis of tumor.The expressions of COX-2 and Ki-67 may be the reference indicators to judge the biological behavious of colorectal carcinoma.

Objective: To investigate the levels of B type natriuretic peptide fibrinogen (BNP) of the systemic infection patients with different cardiac function classifications, and to detect the ventricular structure and the hemodynamics and cardiac function of the patients by transthoracic echocardiography,and to clarify the characteristics of the cardiac function of patients with systemic infection.Methods: The clinical data of 114 patients with systemic infection were collected and analyzed retrospectively.All the patients were divided into heart failure group (n=35) and non-heart failure group (n=79)according to whether or not heart failure occured.The levels of plasma BNP were detected in all the patients,and the values of left ventricular end diastolic diameter (LVIDD) and left ventricular ejection fraction (LVEF) were detected by transthoracic echocardiography.The cardiac function classifications of all the patients were confirmed by the Killip classification method.The correlations between the cardiac function classifications and the levels of plasma BNP and the values of LVEF were analyzed.The diagnostic sensitivity and specificity of the patients with systemic infection complicated with heart failure were analyzed by BNP level detection,transthoracic echocardiography, and BNP level detection combined with transthoracic echocardiography, respectively.Results: Compared with non-heart failure group,the plasma BNP levels and LVIDD values of the patients in heart failure group were obviously increased(t=8.985,P=0.039; t=11.459, P=0.028),but the LVEF values were obviously decreased (t=9.852, P=0.031). The Killip classification of cardiac function had positive relationship with the plasma BNP levels in the patients with systemic infection (r=0.90,P<0.05); the Killip classification of cardiac function had negative relationship with the LVEF values in the patients with systemic infection (r=-0.785,P<0.05).The diagnostic sensitivity and specificity of the patients with systemic infection complicated with heart failure detected by BNP detection combined with transthoracic echocardiography were higher than those detected by other two detection methods alone(P<0.05).Conclusion: The diagnostic sensitivity and specificity of patients with systemic infection complicated with heart failure can be obviously improved by BNP detection combined with transthoracic echocardiography.

Objective: To compare the clinical characteristics between the systemic lupus erythematosus (SLE) patients with positive human cytomegalovirus (HCMV)-IgM antibody and negative HCMV-IgM antibody, and to provide basis for the clinical diagnosis and treatment of SLE.Methods: A total of 63 SLE patients with positive HCMV-IgM and 66 SLE patients with negative HCMV-IgM were selected.The patients who had never used corticosteroid hormone or immunosuppressive drugs were included in initial treatment group(n=48),while those who used either of them were included in retreatment group(n=81).The incidence of clinical manifestations,SLE disease activity index (SLEDAI), absolute counts of CD4⁺ T and CD8⁺T lymphocytes,and CD4⁺/CD8⁺ ratio of the patients in two groups were compared.Results: The anemia,fever,liver dysfunction,leukopenia, and rash were the most common clinical manifestations of the SLE patients.The incidence rates of rash,arthritis,serositis and renal dysfunction and SLEDAI in positive HCMV-IgM group were higher than those in negative HCMV-IgM group (P<0.05). Compared with negative HCMV-IgM group,the absolute count of CD4⁺T lymphocytes was decreased,and the absolute count of CD8⁺ T lymhocytes was increased in positive HCMV-IgM group (P>0.05). The incidence rates of rash and arthritis of the patients with positive HCMV-IgM in initial treatment group were higher than those patients with negative HCMV-IgM (P<0.05). The incidence rates of clinical manifestations of the patients with positive HCMV-IgM and with negative HCMV-IgM in retreatment group had no significant differences(P>0.05).Conclusion: The clinical manifestations of the SLE patients with positive and negative HCMV-IgM are different,and the manifestations of patients with positive HCMV-IgM are more serious.

Objective: To observe the curative effects of modified constraint-induced movement therapy(CIMT) and task-oriented occupation therapy(OT) in the rehabilitation of activity of daily living (ADL) in the patients with sub-acute stroke,and to discuss whether the modified CIMT was suitable for the improvment of ADL.Methods: A total of 90 sub-acute stroke patients were divided into CIMT group(n=48) and control group(n=42),and there were 10 patients dripped out during the study,so there were 42 patients in CIMT group and 38 patients in control group at last.The patients in CIMT group were treated with CIMT,and the patients in control group were treated with task-oriented OT.The scores of Wolf Motor Function Test(WMFT),Fugl-Meyer assesment (FMA) and Barthel index(BI) of the patients in two groups were assessed before treatment and 21 d after treatment,and 30 d after the end of treatment.Results: Compared with before treatment,the WMFT,FMA, and BI scores of the patients in CIMT and control groups after treatment were improved significantly (P<0.05).Compared with control group,the WMFT, FMA,and BI scores of the patients 21 d after treatment in CIMT group were improved (P<0.05).Compared with 21 d after treatment,the WMFT and BI scores of the patients in CIMT group were increased 30 d after the end of treatment (P<0.05),but the FMA score had no change(P>0.05); there were no changes of WMFT,FMA,and BI scores of the patients in control group(P>0.05).Conclusion: The curative effect of modified CIMT in the rehabilitation of ADL in the patients with sub-acute-stroke is superior to task-oriented OT.CIMT could be applied in rehabilitation of the sub-acute stroke patients.

Objective: To compare the curative effects of laparoscopic pyelolithotomy combined with flexible cystoscopy (LPLFC) and percutaneous nephrolithotomy (PCNL) in treatment of complex renal calculi, and to evaluate its safety.Methods: A total of 62 patients with complex renal calculi were randomly divided into LPLFC group (n=31,treated with LPLFC) and PCNL group(n=31,treated with PCNL). The operative time, the hospitalization time,the decreased values of postoperative hemoglobin,occurrence of complications and one-stage stone clearance rates of the patients in two groups were compared.Results: All the operations were finished successfully.A total of 8 patients in LPLFC group complicated with homolateral adrenal or renal diseases received synchronic laparosopic surgery. The operative time and the decreased value of postoperative hemoglobin of the patients in LPLFC group were significantly shorter and lower than those in PCNL group(t=18.3,P<0.01;t=33.2,P<0.01). A total of 5 patients(16.1%) in PCNL group required transfusion postopratively because of blood loss. The hopitaliztion time and one-stage stone clearance rate of the patients in two groups had no significant differences (t=21.2,P=0.7;t=21.9,P=0.71).Conclusion: LPLFC is a minimally invasive operation method with less blood loss, shorter operative time and higher operative safety in treatment of complex renal calculi.

Objective: To explore the clinical features of the patients with Kartagener syndrome(KS), and to review the associated literaturesabout its diagnosis and treatment, and to improve the understanding of the clinician for KS.Methods: The clinical data of one KS patient was collected and the therapeutic effect was analyzed.Results: The patient was clearly diagnoed as KS after relevant examinations.The symptoms such as cough,expectoration,and shortness of breath of the patient were improved after symptomatic treatment,such as anti-inflammation,relieving cough and reducing sputum,and spasmolysis.Conclusion: KS is a congenital autosomal recessive disease.The clinical treatment can only relieve the symptoms,but the structural change is irreversible.The genetics of KS needs further study to reduce its occurrence.

Objective: To explore the clinical characteristics,laboratory indicator features,treatment,and prognosis of the patients with adult-onset Still's disease(AOSD) complicated with hemophagocytic syndrome(HPS),and to provide basis for its diagnosis and treatment.Methods: The clinical data including age,gender,peak of temperature,rash,joint swelling,polyarthralgia,lymphadenopathy,hepatosplenomegaly,and the treatment and prognosis,and the laboratory data including the absolute value of peripheral white blood cells (WBC),neutrophills (NE#),lymphocytes (LY#),monocytes (MO#) and red blood cells (RBC),hemoglobin (HGB),platelet (PLT),aspartate aminotransferase (AST),alanine aminotransferase (ALT), glutamyl-transpeptidase (GGT),alkaline phosphatase (ALP),cholinesterase (CHE),serum ferritin (SF),fibrinogen (FBG),myelogram of three patients with AOSD complicated with HPS were collected and retrospectively analyzed.Results: The general situation showed that all of three patients were female aged over 50 years old. The onset time of HPS was 1 month,1 month and 5 months after AOSD,respectively. All the patients presented fever,rash,and arthritis as the initial symptoms without hepatomegaly or splenomegaly.Only No.3 patient presented lymphadenophy at the beginning of AOSD.After glucocorticoid treatment,the symptoms such as fever,rash,arthritis,and lymphadenophy were improved.The initial symptom of AOSD patient after complicated with HPS was fever,and the temperature was over 39℃.Two patients presented jaundice,no one presented hepatomegaly,splenomegaly or lymphadenophy.The laboratory test results showed that the cytopenia and presistently increasing SF were the early sensitive indicators of AOSD complicated with HPS.The changes of liver enzymes and FBG level followed behind the changes of blood cells and SF level. The phagocytosis was found when bone marrow puncture was performed at the early stage of cytopenia.Conclusion: The clinical characteristic of AOSD patients complicated with HPS is an unexplainedly,recurrently, and continuously high fever,and the characteristics of laboratory test are cytopenia(especially thrombocyopenia),the persistently increasing SF,severe liver injury(especially the increased ALT level),hypofibrinogenemia, and phagocytosis found in bone marrow puncture. The AOSD patients should be early diagnosed and treated in order to reduce the relapse rate and mortality rate.

Objective: To analyze the clinical materials of one patient with flail arm syndrome (FAS), and to provide evidence for its clinical diagnosis and treatment.Methods: The neurological examination,MRI of cranial,cervical and lumbar, and electromyogram (EMG) were performed in the patient and the patient was clinically diagnosed as FAS; the FAS patient was given the treatment to improve the circulation and nourish the nerves.At the same time, comparative analysis of the results between this case report and other case reports of FAS was performed.Results: The patient was diagnosed as FAS after neurological examination and accessory examination:after symptomatic treatment,the condition of patient had the continuous progress,and the upper extremities of the patient were heavier than those of the lower extremities.The patient died of phlegm blocking airway 9 months after discharged.Conclusion: The patients with bilateral symmetric proximal upper extremities weakness and atrophy should be considered as FAS after the other factors are excluded by EMG and imaging examinations.

Objective: To compare the therapeutic effects between primary closue and delayed closure in the patients with dog bite wound,and to discuss the feasibility of primary closure.Methods: A total of 31 patients with dog bite wound were included. The patients were divided into primary closure group(n=20) and delayed closure group (n=11) according to the closue methods. The patients in primary closure group were given the routine soapy water to wash the wound, skin disinfection, local anesthesia, debridement, gentamicin and metronidazole douche, flap transfer and wound closue, antibiotics after the operation, and vaccination of rabies vaccine and tetanus vaccine. The patients in delayed closue group were given the conventional soapy water to wash the wound, skin disinfection, local anesthesia, debridement, gentamicin and metronidazole douche, sterile gauze to cover the wound, regular dressing replacement, delayed closure of wound,routine injection of rabies vaccine and tetanus vaccine after the operation,and systemic antibiotics. The infection rate of wound and Vancouver Scar Rating Scale (VSS) scores of the patients in two groups were compared.Results: Among 20 patients in primary closure group,there were 2 patients (10.00%) showed different degrees of wound infection,and there was no systemic infection, rabies and tetanus; among 11 patients in delayed closure group,there were 2 cases (9.09%) showed different degrees of wound infection,and there was no systemic infection, rabies and tetanus. There were no significant differences in the infection rates of wound of the patients between two groups (P>0.05). The VSS score of the patients in primary closure group (Z=257) was higher than that in delayed closure group (Z=239), and the difference was statistically significant (P<0.05).Conclusion: The infection rates of wound of the patients in primary closure group and delayed closure group are basically the same. However, the wound treated with the primary closure shows cosmetic appearance compared with delayed closure. Therefore,primary closure may be an acceptable therapeutic method for the patients with dog bite wound.

Objective: To observe the efficacy,security,and compliance of new hybrid fusion technology in the treatment of adult degenerative scoliosis (ADS),and to provide the basis for its clinical application.Methods: A total of 50 cases of ADS patients were divided into observation group(n=25) and control group(n=25).The patients in observation group were treated with new hybrid fusion technology,and the patients in control group were treated with traditional open surgery. The operation time,blood loss during the operation,surgical adherence,incidence of complications,improvement rate of lumbar function and fusion rate of bone graft of the patients in two groups were compared.Results: The operation time of patients in observation group was shorter than that in control group (t=6.199,P=6.199); the blood loss during the operation was less than that in control group (t=6.242,P=6.242). The total clinical effective rates of the patients in two groups had no statistically significant difference (χ²=0.758,P=0.758).The incidence of complications of the patients in observation group was lower than that those in control group (χ²=4.667,P=4.667), but the compliance rate were higher than that in control group (χ²=5.357,P=5.357).After treated for 1 and 3 months, the improvement rates of lumbar function and fusion rates of bone graft of the patients in two groups had no statistically significant differences (χ²=1.471,P=0.225; χ²=0.725,P=0.395; χ²=1.587,P=0.208; χ²=0.739,P=0.390);after treated for 6 and 12 months,the above data of the patients in observation group were higher than those in control group (χ²=4.500,P=0.034; χ²=4.667,P=0.031; χ²=5.357,P=0.021; χ²=7.018,P=0.008).Conclusion: New hybrid fusion technology used in ADS treatment has high security,good adherence,shorter operation time,less blood loss and other advantages, and it is suitable for clinical promotion.

Objective: To observe the curative effect of extracorporeal shock wave therapy in the treatment of the patients with bone marrow edema syndrome(BMES) at different positions of lower limbs,and to provide a new and effective method for treatment of BMES.Methods: A total of 39 patients with BMES at different positions of lower limbs were selected. All the patients were treated with extracorporeal focused shock wave by HK-ESWO-AJⅡ apparatus at surface projection of BMES. The voltage was 7-10 kV. The frequency was 60 min^-1. The intensity was 0.18-0.25 mJ·mm^-2. Every treatment point was treated with 1 000 times, once per 5 d for 4 courses. All the patients were told to reduce weight-bearing during the therapy. The visual analogue score(VAS) was used to detect the pain status of the patients;the joint function score was used to detect the joint funtion of lower limbs of the patients;the function independent measure(FIM) index was used to evaluate the ability to live independently of the patients.Results: Compared with before treatment, the VAS score of patients was decreased(P<0.05) and the joint function score and FIM index were increased after treatment(P<0.05). A total of 2 patients were found BMES recurrence during 12-24 months follow-up; one patients occurred at femur head and neck and another patient occurred at distal femur. Both of them were recovery after 4 times of shock wave therapy.There was no patient suffered from osteonecrosis. Conclusion: Extracorporeal focused shock wave therapy could improve the pain symptom and bone marrow edema signs of the BMES patients. Although BMES of lower limbs has recurrence tendency, there is no obvious trend of osteonecrosis and other diseases.

Objective: To investigate the changes of signals in tumor tissue of the tumor-bearing nude mice after SPIO-shRNA molecular probe injection at different time points,and to clarify the mechanism and optimal scanning time of magnetic resonance imaging (MRI) by molecular probe in vivo. Methods: The injection dose of probe was 18 mg·kg^-1. A total of 30 BALB/c tumor-bearing nude mice were randomly divided into six groups(n=5). The mice in each group received MRI examination before and after injected with SPIO-shRNA molecular probe at different time points (12,24,27,30,and 36 h), and the changes of signal intensities in tumor tissue and controlateral muscle tissue of the nude mice were measured at different time points.Then the mice were sacrified after each examination,and the tumor and contralateral muscle tissues were obtained for HE and prussian blue staining, and the morphology of tumor tissue of the nude mice was observed under light microscope.Results: Compared with before injection,the signal intensities of T1WI in tumor tissue were increased at 12,24 and 27 h after injection of molecular probe(P<0.05),while the signal intensities of T2WI and T2*GRE were decreased at 12,24,27,30 and 36 h after injection of molecular probe(P < 0.05 or P < 0.01). The signal intensity of T2WI at 27 h after injection was the highest.However,the signal intensities of T2WI and T2*GRE at 27 h after injection were lower than those at 12,24,30, and 36 h(P<0.05). Compared with before injection,the signal intensity in contralteral muscle tissue of the nude mice at 24 h after injection was decreased(P<0.05). The HE staining results indicated that the structure of tumor tissue was abnormal and the unclear atypia was obvious.The prussian blue staining results showed that the blue-stained iron particles were distributed dispersedly at different time points, and the number of blue-stained iron particles was the most at 27 h after injection.Conclusion: The tumor tissue could be well labeled with the SPIO-shRNA molecular probes,and the optimal MRI scanning time is at 27 h after probe injection.

Objective: To explore the application of image-guided radiotherapy technology(IGRT) in intensity modulated radiation therapy(IMRT) in the patients with rectal cancer before operation,and to analyze its guiding significance for target region.Methods: A total of 67 patients with rectal cancer(Ⅱ-Ⅲ stages,CT3-4N0M0 or CT1-4N1-2M0) were treated with IMRT before operation. IGRT was used for once a week during the radiotherapy and two sets of image contrast errors in the three-dimensional space were obtained.The systematic (Σ) and random (σ) setup errors were calculated.The MPTV was calculated according to the van Herk formula.Results: A total of 389 CBCT scans were analyzed. The error range in the directions of e x, y, z axis were (0.86--1.13), (1.94--2.11),and (1.87--1.81) cm, respectively. The translation errors of the corrected y axis and z axis were less than those before correction (t=3.541, P=0.001; t=3.275, P=0.000); the rotation errors of the corrected x axis and y axis were less than those before correction (t=1.260, P=0.000; t=2.797, P=0.000). Before correction, the MPTV in the directions of x, y, z axis were 2.00, 3.25, and 2.88 cm, respectively, which were 0.73, 1.74, and 1.50 cm after correction, reduced by 1.27, 1.51, and 1.38 cm, respectively.Conclusion: IGRT could effectivly guarantee the accuracy of IMRT of the patients with rectal cancer by reducing the systematic errors and position errors.

Objective: To learn the status of microbial contamination of central water supply in the rural areas of Henan province,and to analysis the influencing factors of water microbial in order to provide basis for making the policy of drinking water safety improvement.Methods: During 2011-2014,the water sampling of central water supply in eight cities (counties) in Xinxiang city of Henan province were selected and the hygiene condition investigation (water source type,water period type,water supply scale,seasonal,local area,and disinfection) was performed.According to the Drinking Water Standard Test Method (GB/T5750-2006),the microorganisms in factory water and tap water were detected,including the total number of colonie,total coliform count,thermotolerant coliform bacteria and Escherichia coli (E.coli).The influencing factors of total number of colonies exceeding the standards were detected by Logistic regression analysis.The influencing factors of total number of colonies and total coliform count were detected by Poisson regression analysis.Results: There were 824,820,526,and 636 water samples in each year during 2011-2014.The most source of supply water was deep water(90%).The samples of drinking water without any treatment accounted for 70.6%. The qualified rates of total number of colonies were 76.0%,75.5%,66.5%,and 72.6%,respectively,and the qualified rates of coliform count were 97.3%,94.9%,99.6%,and 94.3%,respectively.The exceeding standard rates of total number of colonies of disinfected central water supply were lower than those of central water supply without disinfection (P<0.05) during 2011-2014.The exceeding standard rates of total number of colonies of tap water were higher than those of produced water during 2012-2014 (P<0.05).The exceeding standard rates of total number of colonies in medium water supply engineering (100-1 000 t) were lower than those in large(≥ 1 000 t) and small (<100 t) water supply engineerings in 2013 and 2014 (P<0.05).The Logistic regression analysis results showed that the unqualified rate of total number of colonies in wet season was higher than that in dry season (χ²=12.31,P<0.001).The disinfection could reduce the unqualified rate of total number of colonies (χ²=85.32,P<0.001).The unqualified rates of total number of colonies in the engineering for the purpose of removing fluoride and resolving bitter salt water were lower (χ²=34.44,P<0.001;χ²=39.44,P<0.001). The Poisson regression analysis results showed that wet season (χ²=6 972.75,P<0.001),without disinfection (χ²=5 415.46,P<0.001),tap water (χ²=97.64,P<0.001),long operating time (χ²=26.57,P<0.001),low water supply capacity (χ²=4 502.57,P<0.001),the engineering for the purpose of removing fluoride and resolving bitter salt water (χ²=6 226.68,P<0.001;χ²=13 441.50,P<0.001) increased the total number of colonies. The number of E.coli in samples of central water supply was lower in 100-1 000 t of water supply engineerings (χ²=802.18,P<0.001) and the engineering for the purpose of removing fluoride and resolving bitter salt water (χ²=2 711.88,P<0.001;χ²=1 470.91,P<0.001).The number of E.coli in samples of central water supply was higher in the water of wet season (χ²=16 583.10,P<0.001),the water without disinfection (χ²=276.34,P<0.001),tap water (χ²=1 205.26,P<0.001) and the water with long operating time (χ²=3 220.58,P<0.001).Conclusion: The ratio of disinfection of central water supply in the rural areas of Henan provice is low,the qualified rate of colonies is not high,and the qualified rate of total coliform count was high.The microbial contamination in central supply water in the rural areas is influenced by water period type,disinfection,water sample type,operating time,water supply capacity,and purpose of water supply engineering.

Objective: To prepare the polyclonal antibody against growth hormone-releasing hormone receptor splice variant 1 (GHRHR-SV1),and to detect the expressions of GHRHR-SV1 protein in melanoma cells,and to lay a foundation for further study of the pathogenesis of tumor.Methods: The gene sequence of GHRHR-SV1 was designed and synthesized,and then cloned into pET-32 a (+)plasmid to construct the recombinant vector pET-GHRHR-SV1. After identification by PCR and sequencing,the recombinant plasmid was transformed into E.coli BL21 (DE3). Then 0,0.5,1.0,1.5, and 2.0 mmol·L^-1 isopropyl β-D-1-thiogalactopyranoside (IPTG) were added respectively to induce for 1,2,4, and 6 h at 37℃,30℃, and 25℃.The optimal dosage,time and temperature of IPTG of the target protein were observed.The protein was expressed under the optimal condition and purified by affinity chromatography.The polyclonal antibody was obtained by immunizing the rabbit with the purified protein.The titer was assayed by ELISA method.The expression of GHRHR-SV1 protein in B16-F10 cells was detected by Western blotting method.Results: The recombinant vector pET-GHRHR-SV1 was constructed successfully.The expression amount of GHRHR-SV1 protein was the highest when it was induced by 2.0 mmol·L^-1 IPTG for 2 h at 25℃. The relative molecular weight of induced protein was 24 000.After purification,the purity of protein was 80%.The GHRHR-SV1 antibody titer was 1:1 600 000.The GHRHR-SV1 protein was expressed in B16-F10 cells.Conclusion: The recombinant vector pET-GHRHR-SV1 is constructed successfully. The expression amount of GHRHR-SV1 protein in the condition of 2.0 mmol·L^-1 IPTG, 2 h-induction and at 25℃ is the highest.The high titer of ployclonal antibody is obtained.The GHRHR-SV1 protein can express in melanoma cells.

Objective: To establish and evaluate the model of liver regeneration of the cirrhosis mice after partial hepatectomy.Methods: Fifty mice were randomly divided into experiment group(n=25) and control group(n=25). The mice in experiment group were injected with CCl₄ intraperitoneally twice per weekfor 8 weeks and the mice in control group were injected with soya oil.Partial hepatectomy of 37% liver was performed on all mice in two groups. The successful rate of operation, survival rate after operation, survival statue after operation and postoperative liver regeneration rate of the mice in two groups were observed; the pathologic characteristics of liver tissue of the mice in two groups were observed by HE staining and Masson staining;the levels of serum hepatocyte growth factor (HCG) and alanine aminotransferase (ALT) of the mice in two groups were detected by ELASA methods.Results: The successful rates of operation of the mice in two groups were both 100%, and the survival rates of the mice at 6 weeks after operation were 100%.During 3 weeks after operation, the regeneration rates of liver tissue of the mice in experiment group were lower than those in control group; the regeneration rate of liver tissue of the mice in experiment group was equal to that in control group at 3 weeks after operation;the regeneration rates of liver tissue of the mice in experiment group were higher than those in control group from 3 weeks after operation. The HE staining and Masson staining results showed that the liver tissue presented the pathological changes of cirrhosis 8 weeks after injection with CCl₄;after partial hepatectomy, the structures of liver tissue were gradually improved. The levels of serum HGF of the mice in experiment group were significantly higher than those in control group; the levels of serum HGF of the mice in two groups were increased compared with the preoperative levels, and then recovered to the preoperative level. Compared with control group, the levels of serum ALT of the mice in experiment group after hepatectomy were increased significantly, and the duration was longer and the time to reach the peak was delayed.Conclusion: It is extremely easy to establish a stable and reliable liver regeneration model of liver cirrhosis mice after partial hepatectomy by the resection of left lateral lobe of the cirrhosis mice induced by CCl₄ with silk ligation.