Objective To discuss the effect of Sirtuins（Sirt） protein family on the male reproductive system damage cell and animal models induced by bisphenol A （BPA）， and to clarify its effect on the male reproductive system damage induced by BPA. Methods Forty Kunming mice were randomly divided into control group， low dose of BPA group （given 3 mg·kg^－1·d^－1 BPA）， medium dose of BPA group （given 30 mg·kg^－1·d^－1 BPA）， and high dose of BPA group （given 300 mg · kg^－1·d^－1 BPA），and there were 10 mice in each group. The mice in low， medium， and high doses of BPA groups were gavaged with corn oil （0.01 mL·g^－1） mixed with corresponding concentrations of BPA， while the mice in control group were given 0.01 mL·g^－1 corn oil. After 5 weeks， the body weights， testis indexes， and epididymal indexes of the mice in various groups were detected； the sperm qualities of the mice in various groups were detected by using computer assisted semen analysis （CASA） system； the morphology of testis tissue of the mice in various groups was observed by HE staining； the expression levels of Sirt1－Sirt7 proteins in testis tissue of the mice in various groups were detected by Western blotting method. The GC-2 cells were divided into 0， 20， 40， and 80 μmol·L^－1 BPA groups （treated with 0， 20， 40，and 80 μmol·L^－1 BPA）. The proliferation rates of the GC-2 cells in various groups were detected by EdU staining； the percentages of the GC-2 cells at different cell cycles and the apoptotic rates of GC-2 cells in various groups were detected by flow cytometry；the expression levels of Sirt1－Sirt7 proteins in the GC-2 cells in various groups were detected by Western blotting method. Results Compared with control group， the testis index of the mice in high dose of BPA group was decreased （P<0.05）； compared with control group， the percentage of immobile sperm of the mice in high dose of BPA group was increased （P<0.01）， the percentage of rapid progressive sperm was decreased （P<0.01）， the percentage of medium progressive sperm was decreased （P<0.05）， and the deformity rate of the sperm was increased （P<0.01）. The HE staining results showed that the number of spermatogenic cells at all levels in the seminiferous tubules of the mice in different doses of BPA groups showed a decreasing and loosely arranged trend. Compared with control group， the expression level of Sirt6 protein in testis tissue of the mice in low dose of BPA group was decreased （P<0.01）， while the expression levels of Sirt1， Sirt2， and Sirt6 proteins in testis tissue of the mice in medium dose of BPA group mice were decreased （P<0.01），the expression levels of Sirt1， Sirt2， Sirt3， Sirt4， Sirt5， Sirt6， and Sirt7 proteins in testis tissue of the mice in high dose of BPA group were decreased （P<0.05 or P<0.01）. The EdU staining results showed that compared with 0 μ mol·L^－1 BPA group， the proliferation rates of the GC-2 cells in 20， 40， and 80 μmol·L^－1 BPA groups were decreased （P<0.01）. The flow cytometry results showed that after treated for 48 h， compared with 0 μmol·L^－1 BPA group， the apoptotic rates of the GC-2 cells in 20， 40， and 80 μmol·L^－1 BPA groups were increased （P<0.01）. The Western blotting assay results showed that after treated for 24 h， compared with 0 μmol·L^－1 BPA group， the expression levels of Sirt4 and Sirt7 proteins in GC-2 cells of the mice in 20 μmol·L^－1 BPA group were decreased （P<0.05 or P<0.01）， while the expression levels of Sirt4 and Sirt7 proteins in GC-2 cells of the mice in 80 μmol·L^－1 BPA group decreased （P<0.05 or P<0.01）. After treated for 48 h， compared with 0 μmol·L^－1 BPA group， the expression levels of Sirt2， Sirt3， Sirt4， and Sirt6 proteins in GC-2 cells of the mice in 20 μmol·L^－1 BPA group were decreased （P<0.05 or P<0.01）， while the expression levels of Sirt1， Sirt2， Sirt3， Sirt4， and Sirt6 proteins in GC-2 cells of the mice in 40 μmol·L^－1 BPA group were significantly decreased （P<0.05 or P<0.01），and the expression levels of Sirt 5 and Sirt 6 proteins were decreased （P<0.05 or P<0.01）. Conclusion The expression levels of Sirt1－Sirt7 proteins in the male reproductive injury cells and animal models induced by BPA are decreased， which exacerbates the male reproductive system damage induced by BPA.

Objective To discuss the antioxidant capacities of ten edible plant exosomes-like nanoparticles （ELNs） from ginger， green pepper， garlic， mushroom， lemon， yam， grapes， tomatoes， broccoli，and onions in vitro and their protective effects on oxidative injury of the PC12 cells induced by hydrogen peroxide，and to provide the basis for the further study on the ELNs. Methods The PC12 cells were divided into control group， exosome group， hydrogen peroxide group，and hydrogen peroxide+exosomes group. Ten types of ELNs were separated and extracted by using high-speed differential centrifugation. The in vitro antioxidant capacities of these 10 types of ELNs were detected by using 1，1-diphenyl-2-picryl-hydrazyl（DPPH） free radical scavenging system，2，2'-azinobis-（3-ethylbenzthiazoline-6-sulphonate（ABTS） cationic radical system， and ferric ion total reduction antioxidant power（FRAD） system. The viabilities of PC12 cells in various groups were detected by MTT method. Results The DPPH radical scavenging abilities of ten ELNs，from high to low， were mushroom， onion， ginger， lemon， grape， tomato， green pepper， broccoli， yam， and garlic； the capacities of ten ELNs to scavenge ABTS， from high to low， were ginger， mushroom， onion， lemon， yam， grape， green pepper， garlic， tomato， and broccoli； the total antioxidant capacities of ten ELNs in FRAP， from high to low， were ginger， green pepper， onion， mushroom， lemon， broccoli， grape， yam， tomato，and garlic. The antioxidant capacities of five ELNs with stronger antioxidant capacities were increased with the increasing concentration of the ELNs in the DPPH radical scavenging system， ABTS cationic radical system and FRAP system. The half maximal inhibitory concentration（IC₅₀） values of onion were 73.15， 123.02， and 83.00 g·L^－1， the IC₅₀ values of ginger were 124.07， 91.24， and 91.24 g·L^－1， the IC₅₀ values of mushroom were 310.44， 518.04，and 237.10 g·L^－1， the IC₅₀ of green pepper were 969.06， 847.32，and 237.10 g·L^－1 and the IC₅₀ values of lemon were 1 718.94， 544.38，and 962.12 g·L^－1； compared with control group， the viability of the PC12 cells in hydrogen peroxide group was decreased by about 30%. Compared with hydrogen peroxide group， the viability of the PC12 cells in hydrogen peroxide+ELNs groups （green pepper， lemon， yam， and broccoli） were increased by 21.08%， 26.2%， 11.72%， and 15.15%. Conclusion The ELNs from ginger， mushroom， lemon， and onion show stronger antioxidant capacities in the DPPH radical scavenging system， ABTS cationic radical system and FRAP system. The ELNs from green pepper， lemon， yam， and broccoli have protective effects on the oxidative damage of the PC12 cells induced by hydrogen peroxide.

Objective To observe the effect of sulfur dioxide （SO₂） on the myocardial fibrosis（MF） after acute myocardial ischemic injury in the rats， and to explore its mechanism. Methods Twenty-four male SD rats were randomly divided into control group （untreated）， isoproterenol （ISO） group （treated with ISO）， ISO+ SO₂ group （treated with ISO and SO₂）， and SO₂ group （treated with SO₂）， and there were six rats in each group. The acute myocardial ischemic injury model rats in ISO group and ISO+ SO₂ group were induced by intraperitoneal injection with a high dose of ISO （50 mg·kg^－1·d^－1） for 2 d. After successful modeling， the rats in ISO+ SO₂ group and SO₂ group were treated with Na₂SO₃ solution （0.54 mmol·kg^－1·d^－1） and NaHSO₃ solution （0.18 mmol·kg^－1·d^－1） for four weeks， while the rats in control group and ISO group were treated with equal volume of physiological saline intraperitoneally. The levels of myocardial troponin in plasma and electrocardiogram of the rats in various groups were detected；the left ventricular fractional shortening （LVFS） and left ventricular ejection fraction （LVEF） of the rats in various groups were detected by echocardiography；the collage deposition in myocadium tissue of the rats in various groups was detected by Masson staining and the collagen volume fraction （CVF） was calculated； the apoptotic rates of cardiomyocytes of the rats in various groups were detected by Tunel staining；the expression levels of autophagy-related proteins 3 （Atg3）， autophagy-related protein 5 （Atg5）， autophagy-related protein 16L1 （Atg16L1），aspartate proteolytic enzyme containing cysteine 3（Caspase-3）， aspartate proteolytic enzyme containing cysteine 9（Caspase-9）， matrix metalloproteinase 8 （MMP-8）， and tissue inhibitor of metalloproteinase 2 （TIMP-2） proteins in myocardium tissue of the rats in various groups were detected by Western blotting method. Results Compared with control group， the ST segment in electrocardiogram of the rats in ISO group and ISO+ SO₂ group showed elevation and the levels of myocardial troponin were increased （P<0.05），which indicated the acute myocardial ischemic injury models of the rats were established successfully. Compared with control group， the LVEF and LVFS of the rats in ISO group were decreased （P<0.05）， the CVF in myocadium tissue was increased （P<0.05）， the apoptotic rate of the cardiomyocytes was increased （P<0.05）， and the expression levels of Atg3， Atg5， Atg16L1， Caspase-3， Caspase-9， and MMP-8 proteins in myocardium tissue were increased（P<0.05）， and the expression level of TIMP-2 in myocardium tissue was decreased （P<0.05）. Compared with ISO group， the LVEF and LVFS of the rats in ISO+ SO₂ group was increased （P<0.05）， the CVF in myocadium tissue was decreased （P<0.05）， the apoptotic rate of the cardiomyocytes was decreased （P<0.05）， the expression levels of Atg3， Atg5， Atg16L1， Caspase-3， Caspase-9， and MMP-8 proteins in myocardium tissue were decreased（P<0.05）， and the expression level of TIMP-2 in myocardial tissue was increased （P<0.05）. Conclusion SO₂ can improve the MF after acute myocardial ischemic injury in the rats， and its mechanism may be related to the inhibition of excessive myocardial cell autophagy and reduction of the apoptosis of the cardiomyocytes.

Objective To discuss the effect of microRNA-491-5p（miR-491-5p） over-expression on the proliferation and migration of the human nasopharyngeal carcinoma HONE-1 cells， and to provide the evidence for the study of the pathogenesis and targeted therapy of nasopharyngeal carcinoma. Methods The human nasopharyngeal carcinoma HONE-1 cells were divided into control group （transfected with control plasmid） and miR-491-5p group （transfected with miR-491-5p plasmid）. The transfection efficiencies of the HONE-1 cells in two groups were observed under fluorescence microscope； the proliferation activities of the HONE-1 cells in two groups were detected by CCK-8 assay； the number of migration HONE-1 cells in two groups were detected by Transwell chamber assay；the expression levels of Vimentin and E-cadherin mRNA in the HONE-1 cells in two groups were detected by real-time fluorescence quantitative PCR（RT-qPCR）method； the expression levels of Vimentin and E-cadherin proteins in the HONE-1 cells in two groups were detected by Western blotting method. Results Compared with control group， the proliferation activities（t=2.832， P=0.0473； t=4.522， P=0.0014； t=9.308， P<0.01） and number of migration HONE-1 cells （t=9.639， P<0.01） in miR-491-5p group at 24，48，and 72 h after treatment were significantly decreased； compared with control group， the expression levels of Vimentin mRNA （t=7.535， P=0.0017） and protein （t=7.219， P=0.0187） in the HONE-1 cells in miR-491-5p group were significantly decreased， and the expression levels of E-cadherin mRNA （t=4.88， P=0.0395） and protein （t=5.754， P=0.0289） were significantly increased. Conclusion Over-expression of miR-491-5p can inhibit the proliferation， migration， and epithelial-mesenchymal transition （EMT） of the human nasopharyngeal carcinoma cells.

Objective To observe the changes of inflammation responses induced by lipopolysaccharide （LPS） in the single cultured Müller cells system and co-culture system of Müller cells and microglia of the mice， and to elucidate the interaction between the Müller cells and the microglia. Methods The Müller cells QMMuC-1 and microglia BV2 were cultured， and immunofluorescence staining was used to observe the morphology of two kinds of cells. The experiment was divided into single-culture control group ［QMMuC-1 cells cultured alone， treated with phosphate buffer saline（PBS）］， co-culture control group （QMMuC-1 cells and BV2 cells co-cultured with the ratio of 1∶1， treated with PBS buffer）， single-culture experimental group （QMMuC-1 cells cultured alone， treated with 10 mg·L^－1 LPS）， and co-culture experimental group （QMMuC-1 cells and BV2 cells co-cultured， treated with 10 mg·L^－1 LPS）. Immunofluorescence staining was used to observe the levels of glial fibrillary acidic protein （GFAP） in the cells in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） mRNA in the QMMuC-1 cells in various groups. Results The expressions of glutamine synthetase （GS） and GFAP in the QMMuC-1 cells were positive， and the expression of ionized calcium-binding adapter molecule 1 （Iba-1） in the BV2 cells was positive. Compared with single-culture control group， the level of GFAP in the QMMuC-1 cells in single-culture experimental group was increased by 1.7 fold （P=0.005）. Compared with co-culture control group， the level of GFAP in the QMMuC-1 cells in co-culture experimental group was increased by 2 fold （P=0.003），and the morphology of the cells gradually became fusiform. Compared with single-culture experimental group， the level of GFAP in the QMMuC-1 cells in co-culture experimental group was increased by 1.4 fold （P=0.0006）， and most cells exhibited a fusiform shape. Compared with single-culture control group， the expression levels of IL-1β， IL-6， and TNF-α mRNA in the QMMuC-1 cells in single-culture experimental group were increased， but the expression levels had no significant differences（P>0.05）. Compared with co-culture control group， the expression levels of IL-1β， IL-6， and TNF-α mRNA in the QMMuC-1 cells in co-culture experimental group were increased （P<0.05）. Compared with single-culture experimental group， the expression levels of IL-1β and TNF-α mRNA in the QMMuC-1 cells in co-culture experimental group were increased （P<0.05）. Conclusion LPS may induce the release of inflammatory cytokines from the activated microglia， which subsequently act on the Müller cells and exacerbate the inflammatory response in the Müller cells.

Objective To discuss the effect of long non-coding RNA （lncRNA） gastric cancer related transcript 2（GACAT2） gene on the proliferation and stemness of the lung cancer A549 and H1299 cells， and to clarify the effect of lncRNA GACAT2 gene on the occurrence and development of lung cancer. Methods The full-length sequence of lncRNA GACAT2 gene was amplified by real-time fluorescence quantitative PCR （RT-qPCR） method， and was cloned into the pc3.1 （+） eukaryotic expression vector. The construction of GACAT2 over-expression vector was confirmed by bacterial liquid PCR method and gene sequencing. The pc3.1-GACAT2 over-expression plasmid and control plasmid pc3.1 were separately transfected into the human lung cancer A549 and H1299 cells. The A549 and H1299 cells with stable over-expression of GACAT2 were established by G418 screening， and the cells were divided into blank vector group （transfected with pc3.1 empty vector） and pc3.1-GACAT2 group （transfected with pc3.1-GACAT2 recombinant plasmid）. RT-qPCR method was used to detect the expression levels of GACAT2 mRNA in the A549 and H1299 cells in two groups；CCK-8 assay was used to detect the proliferation activities of the cells in two groups； cell clone formation assay was used to detect the clone formation numbers of the cells in two groups； stem cell sphere formation experiment was used to detect the sphere formation numbers of the A549 and H1299 cells in two groups. Results The eukaryotic over-expression vector of GACAT2 was successfully constructed. Compared with blank vector group， the expression levels of GACAT2 mRNA in the A549 and H1299 cells in pc3.1-GACAT2 group were increased （P<0.05 or P<0.01）， the cell proliferation activities were decreased （P<0.05 or P<0.01），the clone formation numbers were decreased （P<0.05 or P<0.01）， and the sphere formation numbers were decreased （P<0.05 or P<0.01）. Conclusion Over-expression of human lncRNA GACAT2 gene can inhibit the proliferation and stemness of the lung cancer A549 and H1299 cells.

Objective To discuss the inductive effect of testosterone on the apoptosis of the granulosa cells in vitro，and to clarify its mechanism. Methods The inhibitory rates of proliferation of human ovarian granulosa cells SVOG were detected by methyl thiazolyl tetrazolium（MTT） assay. After screening the optimal concentrations of the drugs， the SVOG cells were divided into control group， testosterone group （1.0×10^－5 mol·L^－1 testosterone），testosterone+fluorothiamine group（1.0×10^－5 mol·L^－1 testosterone+ 1.0×10^－5 mol·L^－1 fluorothiamine），and testosterone+tauroursodeoxycholic acid （TUDCA）group （1.0×10^－5 mol·L^－1 testosterone+1.5 g·L^－1 TUDCA）. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of spliced X-box binding protein 1 （XBP1s）， activating transcription factor 4 （ATF4）， transcription factor C/EBP homologous protein （CHOP）， and death receptor 5 （DR5） mRNA in the SVOG cells in various groups； flow cytometry was used to detect the apoptotic rates of the SVOG cells in various groups. Results Compared with control group， the SVOG cells in testosterone group showed morphological changes， the number of atypical cells was increased， the granularity in the cytoplasm was increased， the cell fragments in the culture medium was increased， and the inhibitory rate of proliferation of the SVOG cells was increased （P<0.05）. Compared with testosterone group， the morphological changes of the cells in testosterone +fluorothiamine group and testosterone+TUDCA group were decreased and inhibitory rates of proliferation of the cells were decreased（P<0.05）. The flow cytometry results showed that compared with control group， the apoptotic rate of the SVOG cells in testosterone group was significantly increased （P<0.05）. Compared with testosterone group， the apoptotic rates of the SVOG cells in testosterone + TUDCA group and testosterone + fluorothiamine group were significantly decreased （P<0.05）. The RT-qPCR analysis results showed that after treated for 48 h， compared with control group， the expression levels of XBP1s， ATF4， CHOP， and DR5 mRNA in the SVOG cells in testosterone group were increased （P<0.05）. Compared with testosterone group， the expression levels of XBP1s，ATF4，CHOP， and DR5 mRNA in the SVOG cells in testosterone+TUDCA group and testosterone+fluorothiamine group were decreased （P<0.05）. Conclusion Testosterone can induce the apoptosis of human ovarian granulosa cells and cause follicular atresia，and its mechanism may be that androgen combined with adrogen receptor（AR） activates the CHOP-DR5 pathway， induces endoplasmic reticulum stress（ERS）， and promotes the apoptosis of the granulosa cells.

Objective To discuss the change of sensitivity of the non-small-cell lung cancer （NSCLC） A549 cells to doxorubicin （Dox） after silencing forkhead protein 3 （FOXP3） gene，and to clarify its mechanism involved in Dox resistance. Methods The human NSCLC A549 cells were transfected with FOXP3 small interfering RNA（siRNA ）by lipofectamine method. The cells were divided into blank control group （without transfection）， si-NC group （transfected with control-siRNA）， and si-FOXP3 group （transfected with FOXP3-siRNA）. Western blotting and immunofluorescence methods were used to detect the expression levels of FOXP3 protein in the A549 cells in various groups；the proliferation activities and half-maximal inhibitory concentration （IC₅₀） values of the A549 cells in various groups were detected by CCK-8 method. The A549 cells were treated with 0， 10， and 20 μmol·L^－1 DAPT， and regarded as 0， 10， and 20 μmol·L^－1 DAPT groups， respectively. Additionally， the A549 cells were treated with 0 μmol·L^－1 DAPT， 1.0 mg·L^－1 Dox， and 1.0 mg·L^－1 Dox combined with 10 μmol·L^－1 DAPT， and regarded as 0 μmol·L^－1 DAPT， 1.0 mg·L^－1 Dox， and 1.0 mg·L^－1 Dox combined with 10 μmol·L^－1 DAPT groups， respectively. Western blotting method was used to detect the expression levels of Notch1，Hes1， and FOXP3 proteins in the A549 cells in various groups； the IC₅₀ values and expression levels of Notch1， Hes1， and FOXP3 proteins in the A549 cells in 0， 10， and 20 μmol·L^－1 DAPT groups were detected by CCK-8 and Western blotting methods；the expression levels of FOXP3， P-glycoprotein（P-gp）， Notch1， and Hes1 proteins in the A549 cells in 0 μmol·L^－1 DAPT， 1.0 mg·L^－1 Dox， and 1.0 mg·L^－1 Dox combined with 10 μmol·L^－1 DAPT groups were detected by Western blotting method. Results Compared with blank control and si-NC groups， the expression level of FOXP3 protein in the A549 cells in si-FOXP3 group was significantly decreased （P<0.01）， the proliferative activity and IC₅₀ value were decreased （P<0.05 or P<0.01），and the expression levels of Notch1，Hes1 and FOXP3 proteins were significantly decreased （P<0.01）. Compared with 0 μmol·L^－1 DAPT group， the expression levels of Notch1， Hes1， and FOXP3 proteins in the A549 cells in 10 and 20 μmol·L^－1 DAPT groups were significantly decreased （P<0.01）. Compared with 0 μmol·L^－1 DAPT group， the expression levels of FOXP3， P-gp， Notch1， and Hes1 proteins in the A549 cells in 1.0 mg·L^－1 Dox group were significantly increased （P<0.01）. Compared with 1.0 mg·L^－1 Dox group， the expression levels of FOXP3， P-gp， Notch1， and Hes1 proteins in the A549 cells in 1.0 mg·L^－1 Dox combined with 10 μmol·L^－1 DAPT group were significantly decreased （P<0.01）. Conclusion Silencing FOXP3 can enhance the sensitivity of the NSCLC cells to Dox， and its mechanism is related to the inhibition of the Notch1/Hes1 signaling pathway.

Objective To discuss the protective effect of ginsenoside Rh2 on kidney injury in the rats with diabetic kidney lesions （DKL），and to clarify its mechanism. Methods The DKL rat model was established by using the high-fat and high-energy diet combined with intraperitoneal injection of streptozotocin（STZ）.Thirty SD model rats were randomly divided into model group， low dose （10 mg·kg^－1） of Rh2 group and high dose （20 mg·kg^－1） of Rh2 group， and there were 10 rats in each group；other ten SD rats were regared as normal control group.The body weights and kidney weights of the rats in various groups were detected，and the kidney index was caculated；after intervented for 8 weeks， automatic analyzer was used to detect the levels of serum creatinine （Scr）， urea nitrogen （BUN） and 24 h urinary protein （UP）in serum of the rats in various groups；enzyme linked immunosorbent assay （ELISA） method was used to detect the levels of collagen type Ⅳ（Col Ⅳ） in kidney tissue of the rats in various groups； Western blotting method was used to detect the expression levels of transforming growth factor-β1 （TGF-β1），Smad3 and Smad7 proteins in kidney tissue of the rats in various groups. Results Compared with normal control group， the body weight of the rats in model group was decreased（P<0.01），the kidney weight and kidney index were significantly increased （P<0.01）， the levels of Scr， BUN and 24 h UP in serum were increased （P<0.01）， the level of Col Ⅳ in kidney tissue was increased（P<0.01）， the expression levels of TGF-β1 and Smad3 proteins in kidney tissue were increased（P<0.01）， and the expression level of Smad7 protein in renal tissue was decreased （P<0.01）. Compared with model group，the body weight of the rats in low and high doses of Rh2 groups were inecreased （P<0.05 or P<0.01）， the kidney weight and kidney index were decreased （P<0.05 or P<0.01），the levels of Scr， BUN and 24 h UP in serum were significantly decreased （P<0.05 or P<0.01）， the level of Col Ⅳ in kidney tissue was decreased（P<0.01）， the expression levels of TGF-β1 and Smad3 proteins in kidney tissue were decreased（P<0.01）， and the expression level of Smad7 protein in kidney tissue was increased （P<0.01）. Conclusion Ginsenoside Rh2 can inhibit the kidney fibrosis and improves the kidney function in the DKL rats， and has a protective effect on the kidney of DKL rats； its mechanism may be related to regulating the expression of TGF-β1/Smads signaling pathway.

Objective To discuss the effect of salidroside on the apoptosis of the bone marrow CD71+ nucleated red blood cells（RBC） in the rat model of high altitude polycythemia （HAPC），and to clarify its mechanism. Methods orty-five male SD rats were randomly divided into control group （exposed to 1 500 m altitude + NaCl solution）， model group （exposed to 5 000 m altitude + NaCl solution）， and drug group （exposed to 5 000 m altitude + 0.15 g·kg^－1·d^－1 salidroside），and there were 15 rats in each group. The RBC counts， hemoglobin （Hb） levels， and hematocrit （HCT） levels of the rats in various groups were detected； flow cytometry was used to detect the percentages of bone marrow CD71+nucleated RBC， apoptotic rates of bone marrow CD71+nucleated RBC， levels of mitochondrial membrane potential （MMP） and expression levels of cysteinyl aspartate-specific protease-3 （Caspase-3） in bone marrow CD71+nucleated RBC of the rats in various groups； Western blotting method was used to detect the expression levels of B-cell lymphoma-2 （Bcl-2）， Bcl2-associated X protein （Bax） ，and cysteinyl aspartate-specific protease-9 （Caspase-9） proteins in bone marrow CD71+nucleated RBC of the rats in various groups. Results Compared with control group， the RBC count， Hb level， HCT level of the rats in model group were increased （P<0.01）， and the percentage and apoptotic rate of bone marrow CD71+nucleated RBC were increased （P<0.01）， and the expression levels of Bax， Caspase-9， and Caspase-3 proteins in the bone marrow CD71+nucleated RBC were increased （P<0.05）. Compared with model group， the percentage of bone marrow CD71+nucleated RBC of the rats in drug group was decreased （P<0.01）， the apoptotic rate of bone marrow CD71+nucleated RBC（P<0.01）， and the MMP level in the bone marrow CD71+nucleated RBC were increased （P<0.05）， and the expression levels of Caspase-3， Bax， and Caspase-9 proteins in the bone marrow CD71+nucleated RBC were decreased （P<0.01）. Conclusion Salidroside can effectively inhibit the increasing of apoptosis of nucleated RBC in bone marrow of the HAPC rats and improve the excessive accumulation of the RBC caused by hypoxia， which may have a certain preventive and protective effect in the pathogenesis of HAPC in the rats.

Objective To discuss the effect of sodium cromoglycate （SCG） on the neural function and cognitive function in the rats with cerebral ischemic stroke （CIS）， and to clarify its mechanism. Methods A total of 80 male SD rats were selected and 18 were randomly selected as sham operation group. The remaining rats were constructed the cerebral ischemia/reperfusion（I/R） injury models by thread embolization method. The 45 successful modeling rats were randomly divided into I/R group， I/R + low dose of SCG group， and I/R +high dose of SCG group（n=15）.At 24 h after modeling， the rats in I/R+low dose of SCG group and I/R +high dose of SCG group were intraperitoneally injected with 20 and 60 mg·kg^－1 SCG for 2 weeks，respectively. The neurological scores of the rats in various groups were evaluated by modified neurological severity score（mNSS） criterion； the area of cerebral infarction was detected by using 2，3，5-triphenyltetrazolium chloride （TTC） staining；the Y-maze experiment was used to detect the memory abilities of the rats in various groups；electrophysiological measurement was used to analyze the accurate reaction rate， accurate reaction time， and field excitatory postsynaptic potential （fEPSP） slopes of the rats in various groups；Golgi staining was used to detect the morphology of dendritic spines in dentate gyrus of hippocampus in brain tissue of the rats in various groups；immunofluorescence method was used to detect the numbers of 5-bromodeoxyuridine （BrdU） positive cells and the expression levels of doublecortin （DCX） protein in dentate gyrus of hippocampus in brain tissue of the rats in various groups； Western blotting method was used to detect the expression levels of brain-derived neurotrophic factor （BDNF），tyrosine kinase B receptor （TrkB）， neurotrophin 3 （NT-3），and tyrosine kinase C receptor （TrkC） proteins in brain tissue of the rats in various groups. Results Compared with sham operation group， the neurological score and the cerebral infarction area of the rats in I/R group were increased（P<0.05），the accurate reaction rate was decreased （P<0.05），and the accurate reaction time was increased （P<0.05）， the fEPSP slope and the density of dendritic spine was decreased （P<0.05）， the number of BrdU positive cells in dentate gyrus of hippocampus in brain tissue and the expression level of DCX protein in hippocampus tissue had no significant differences（P>0.05），and the expression level of NT-3 protein in hippocampus tissue was increased（P<0.05），but the expression levels of BNDF，TrkB， and TrkC proteins had no significant differences（P>0.05）. Compared with I/R group， the neurological scores of the rats in I/R+high dose of SCG group was decreased （P<0.05）， the infarction area was decreased （P<0.05）， the accurate reaction rate was increased （P<0.05）， the accurate reaction time was decreased （P<0.05），the fEPSP slope and the dendritic spine density were increased （P<0.05），the number of BrdU positive cells and the expression level of DCX protein in dentate gyrus of hippocampus in the brain tissue were increased （P<0.05）， and the expression levels of BDNF，TrkB， NT-3， and TrkC proteins were significantly increased （P<0.05）. Conclusion SCG has the protective effect on the neurological and cognitive functions in the I/R model rats and its mechanism may be related to the increasing of the expressions of BDNF/TrkB and NT-3/TrkC signaling pathway related proteins and promoting the proliferation of the neurons.

Objective To discuss the inhibitory effect of bone marrow-derived mesenchymal stem cells （BMSCs） over-expressing forkhead box transcription factor 1 （FOXO1） on the eosinophil infiltration and airway remodeling of the asthmatic mice， and to clarify the possible mechanism. Methods The BMSCs of the mice were isolated， and Oil Red O staining and Alizarin staining were used to identify the BMSCs. Flow cytometry was used to identify the phenotypes of the BMSCs. The BMSCs at logarithmic growth phase were collected and divided into control group （without treatment）， FOXO1-BMSCs group （infected with recombinant lentivirus carrying FOXO1 gene）， and NC-BMSCs group （infected with negative control recombinant lentivirus）. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of FOXO1 mRNA in the BMSCs in various groups； Western blotting method was used to detect the expression levels of FOXO1 protein in the BMSCs in various gorups. Forty mice were randomly divided into control group （given saline）， model group （given saline）， NC-BMSCs group （given NC-BMSCs cell suspension）， and FOXO1-BMSCs group （given FOXO1-BMSCs cell suspension），and there were 10 mice in each group. Except for control group， the mice in the other three groups were sensitized with ovalbumin （OVA） and challenged with aerosol to establish the asthma models. The cell classification counts in bronchoalveolar lavage fluid （BALF） of the mice in various groups were detected；the histopathology of lung tissue of the mice in various groups was observed by HE staining；the expressions of α-smooth muscle actin （α-SMA） and proliferating cell nuclear antigen （PCNA） in lung tissue of the mice in various groups were detected by immunofluorescence method； the bronchial lumen circumference （Pi）， wall area （W）， smooth muscle area （S）， and numbers of smooth muscle cell nuclei （N） of the mice in various groups were detected by using Image-Pro Plus Software，and the ratios of S/Pi， W/Pi， and N/Pi were calculated； the expression levels of matrix metalloproteinase-9 （MMP-9），matrix metalloproteinase-12 （MMP-12）， and tissue inhibitor of metalloproteinase-1 （TIMP-1） proteins in lung tissue of the mice in various groups were detected by Western blotting method. Results The isolated and cultured BMSCs showed a spindle-shaped morphology， and significant red lipid droplets and orange-red precipitates could be observed after Oil Red O and Alizarin staining. The expression amonuts of CD29， CD44， and CD71 on surface of the BMSCs were increased， while the expression amonuts of CD34， CD45， and HLA-DR were decreased. Compared with control group and NC-BMSCs group， the expression levels of FOXO1 mRNA and protein in the BMSCs in FOXO1-BMSCs group were increased （P<0.05）. Compared with model group and NC-BMSCs group， the counts of eosinophils， macrophages， lymphocytes， and neutrophils in bronchoalveolar lavage fluid （BALF） of the mice in FOXO1-BMSCs group were decreased（P<0.05）， and the inflammatory cell infiltration in the airways and alveoli was alleviated； the ratios of S/Pi， W/Pi， and N/Pi were decreased（P<0.05）， and the expression amounts of α-SMA and PCNA in lung tissue were decreased， the expression levels of MMP-9， MMP-12， and TIMP-1 proteins in lung tissue were decreased （P<0.05）. Conclusion The BMSCs over-expressing FOXO1 can attenuate the inflammatory cell infiltration， particularly eosinophil infiltration， and inhibit airway remodeling in the asthmatic mice， thus alleviating the asthma symptoms.

Objective To discuss the effect of paeonol on the proliferation， migration and chemokinereceptor4 （CXCR4）/signal transducer and activators of transcription3 （STAT3） pathway of the human osteosarcoma（OS ） MG-63 cells， and to clarify its possible mechanism. Methods The MG-63 cells were cultured in vitro and divided into control group （without drug intervention） and different concentrations of paeonol groups （given 10， 20， 40， 80， 160，and 320 mg·L^－1 paeonol， respectively）.The survival rates of the MG-63 cells in various groups were detected by CCK-8 method and the half inhibitory concentration （IC₅₀） value was selected as the drug concentration for subsequent experiment. The MG-63 cells were divided into control group（without drug treatment） and paeonol group（given 215.8 mg·L^－1 paeonal）.The expression levels of CXCR4， interleukin 6 （IL-6），and phosphorylated STAT3 （p-STAT3），and STAT3 proteins in the MG-63 cells in various groups were detected by Western blotting method.The MG-63 cells were divided into control group， paeonol group， paeonol+empty group （given 215.8 mg·L^－1 paeonol+empty plasmid）， and paeonol+overexpression group （given 215.8 mg·L^－1 paeonol+CXCR4 over-expression plasmid）. The scratch healing rates of the MG-63 cells in various groups were detected by cell scratch experiment， the clone formation rates the MG-63 cells in various groups were detected by cloning formation experiment， and the expression levels of CXCR4， IL-6， p-STAT3， and STAT3 proteins in the MG-63 cells in various groups were detected by Western blotting method. Results Compared with control group， the survival rates of the MG-63 cells treated in 40， 80， 160， and 320 mg·L^－1 paeonol groups were decreased （P<0.05）， and the IC₅₀ value of paeonol was 215.8 mg·L^－1. Compared with control group， the expression levels of CXCR4， IL-6， and p-STAT3 proteins in the MG-63 cells in paeonol group were decreased （P<0.05）. Compared with control group， the scratch healing rates and clone formation rates of the MG-63 cells in paeonol group and paeonol+empty group were decreased （P<0.05）， and the expression levels of CXCR4， IL-6，and p-STAT3 proteins in the MG-63 cells were decreased （P<0.05）； compared with paeonol group，the scratch healing rate， clone formation rate， expression levels of CXCR4， IL-6， p-STAT3，and STAT3 proteins in the MG-63 cells in paeonol+empty group had no significant differences（P>0.05）； compared with paeonol group， the scratch healing rate and clone formation rate of the MG-63 cells in paeonol+overexpression group were increased （P<0.05）， and the expression levels of CXCR4， IL-6 and p-STAT3 proteins were increased （P<0.05）. Conclusion Paeonol can inhibit the proliferation， migration，and clonogenic ability of the MG-63 cells， and its mechanism may be related to the regulation of CXCR4/STAT3 signal pathway.

Objective To discuss the expressions of serum and glucocorticoid-regulated kinase 3 （SGK3） mRNA and protein in the oocytes of the mice at different stages and their subcellular localizations，and to provide the evidence for elucidating the regulatory mechanism of SGK3 in the cell cycle of oocytes of the mice. Methods The oocytes of the mice at the germinal vesicle （GV）stage， germinal vesicle breakdown （GVBD） stage， first meiotic division （MⅠ）stage， and second meiotic division （MⅡ） stage were collected by using superovulation techniques. Real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to detect the expression levels of SGK3 mRNA and protein in the oocytes at different stages； the phosphorylation expressions of Cdc25B-Ser30 in the oocytes at different stages of the mice were detected by Western blotting method after collecting the oocytes at GV， GVBD， and MⅠ stages； the subcellular localization of SGK3 in the oocytes of the mice at different stages was observed by immunofluorescence. Results SGK3 mRNA and protein were expressed in the oocytes of the mice at all stages. Compared with GV stage， the expression levels of SGK3 mRNA and protein in the oocytes at GVBD and MⅡ stages were increased （P<0.01）. Compared with GVBD stage， the expression levels of SGK3 mRNA and protein in the oocytes at MⅠ stage were decreased （P<0.01），and the expression levels of SGK3 mRNA and protein in the oocytes at MⅡ stage were increased （P<0.01）. Compared with MⅠ stage， the expression levels of SGK3 mRNA and protein in the oocytes at MⅡ stage were increased （P<0.01）.The Cdc25B-Ser30 was dephosphorylated at GV stage and phosphorylated at GVBD and MⅠ stages.The SGK3 was localized at the nuclear membrane of the oocytes at GV stage，and entered the nucleus before GVBD stage，and localized at the nucleus during GVBD stage， distributed throughout the oocytes during MⅠ stage， and re-entered the nucleus from the cytoplasm during MⅡ stage. Conclusion SGK3 is dynamically expressed in the oocytes of the mice at different stages， and it exhibits the nuclear-cytoplasmic shuttling. The dynamic changes in the SGK3 localization may be involved in the activation of cell cycle progression.

Objective To discuss the effect of macrophage-derived extracellular vesicle long non-code RNA（lncRNA） highly up-regulated in liver cancer（HULC） on the metastasis of hepatocellular carcinoma （HCC），and to elucidate its mechanism. Methods The bone marrow mononuclear cells were isolated from the mice and differentiated into bone marrow-derived macrophages （BMDM） through phorbol myristate acetate induction. Interleukin-4 （IL-4） was used to induce the M2 macrophage polarization，regarded as tumor-associated macrophages（TAM）.The TAM-derived extracellular vesicles （referred to as TAM-exos） were collected using differential centrifugation. RNA interference was used to transfect TAM with lncRNA HULC siRNA or negative control （NC） plasmids， and the extracellular vesicles secreted by TAM in various groups were extracted （referred to as lncRNA HULC-siRNA-exos and NC-exos）. The HepG2 cells were divided into control group and TAM-exos group； NC-exos group and lncRNA HULC-siRNA-exos group； lncRNA HULC-siRNA-exos group and lncRNA HULC-siRNA-exos+SKL2001 group according to the purpose of the experiment. After the corresponding treatments， Transwell champer assay was used to detect the numbers of migration and invasion HepG2 cells in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression level of lncRNA HULC in the HepG2 cells in various groups； Western blotting method was used to detect the expression levels of Wnt3A， β-catenin， c-Myc， and Cyclin D1 in the HepG2 cells in various groups. The nude mouse orthotopic liver cancer transplantation model was established， and the mice were divided into control group， NC-exos group， TAM-exos group， and lncRNA HULC-siRNA-exos group. The numbers of lung metastatic lesions in the nude mice in various groups after treatment with M2 macrophage-derived extracellular vesicles containing lncRNA HULC were detected. Results The TAM-exos displayed the morphology and biological characteristics of extracellular vesicles. Compared with control group， the numbers of migration and invasion HepG2 cells in TAM-exos group were increased（P<0.05）， and the expression levels of lncRNA HULC， Wnt3A， β-catenin， c-Myc， and Cyclin D1 proteins in the HepG2 cells in TAM-exos group were increased （P<0.05）. Compared with NC-exos group， the numbers of migration and invasion HepG2 cells in lncRNA HULC-siRNA-exos group were decreased，and the expression levels of lncRNA HULC， Wnt3A， β-catenin， c-Myc， and Cyclin D1 proteins in the HepG2 cells were decreased （P<0.05）. Compared with lncRNA HULC-siRNA-exos group， the numbers of migration and invasion HepG2 cells in lncRNA HULC-siRNA-exos+SKL2001 group were increased（P<0.05），and the expression levels of Wnt3A， β-catenin， c-Myc， and Cyclin D1 proteins in the HepG2 cells were increased （P<0.05）. The nude mouse experiment results showed that compared with control group， the number of lung metastatic lesions of the nude mice in TAM-exos group was increased （P<0.05）； compared with TAM-exos group and the NC-exos group， the number of lung metastatic lesions of the nude mice in lncRNA HULC-siRNA-exos group was decreased （P<0.05）. Conclusion The TAM-derived extracellular vesicle lncRNA HULC promotes the invasion and metastasis of the HCC cells， and its mechanism may be associated with the activation of the Wnt signaling pathway.

Objective To discuss the effect of exogenous CXC chemokine ligand 10（CXCL10） on the proliferation and migration of the hepatocellular carcinoma（HCC）SMMC-7721 cells， and to clarify its mechanism. Methods The human HCC SMMC-7721 cells were divided into 0 mg·L^－1 CXCL10 group，10 mg·L^－1 CXCL10 group，and 30 mg·L^－1 CXCL10 group according to the CXCL10 concentration. Some of the above cells were treated with extracellular regulated protein kinase（ERK） inhibitor PD98059 （80 μmol·L^－1）， then the SMMC-7721 cells were divided into 0 mg·L^－1 CXCL10+PD98059 group， 10 mg·L^－1 CXCL10+PD98059 group， and 30 mg·L^－1 CXCL10+PD98059 group. The proliferation rates of the SMMC-7721 cells in various groups were detected by CCK-8 method； the EdU positive expression rates in SMMC-7721 cells in various groups were detected by EdU method； the migration rates of the SMMC-7721 cells in various groups were detected by Transwell chamber assay； the expression levels of ERK， phosphorylated ERK（p-ERK）， and Cyclin D1 proteins in the SMMC-7721 cells in various groups were detected by Western blotting method. Results The CCK-8 results showed that after cultured for 24 h， compared with 0 mg·L^－1 CXCL10 group， the proliferation rates of the cells in 10 mg·L^－1 CXCL10 group and 30 mg·L^－1 CXCL10 group were increased （P<0.05 or P<0.01）；the EdU detection results showed that compared with 0 mg·L^－1CXCL10 group， the positive expression rates of EdU in the cells in 10 mg·L^－1 CXCL10 group and 30 mg·L^－1CXCL10 group were increased （P<0.01）. The Transwell chamber assay results showed that after cultured for 48 h， compared with 0 mg·L^－1 CXCL10 group， the migration rates of the cells in 10 mg·L^－1CXCL10 group and 30 mg·L^－1 CXCL10 group were increased （P<0.01）.The Western blotting results showed that after cultured for 24 h and treated with CXCL10 for 24 h，compared with 0 mg·L^－1 CXCL10 group， the expression levels of ERK， p-ERK，and Cyclin D1 proteins in the SMMC-7721 cells in 10 mg·L^－1 CXCL10 group and 30 mg·L^－1 CXCL10 group were increased （P<0.05）. The CCK-8 method results showed that after treated with ERK inhibitor PD98059， compared with 0 mg·L^－1 CXCL10 group， the proliferation rates of the SMMC-7721 cells in 10 mg·L^－1 CXCL10+PD98059 group and 30 mg·L^－1 CXCL10+PD98059 group were decreased （P<0.05）； compared with 10 mg·L^－1 CXCL10 group， the proliferation rate of the SMMC-7721 cells in 10 mg·L^－1 CXCL10+PD98059 group was decreased （P<0.05）； compared with 30 mg·L^－1 CXCL10 group， the proliferation rate of the SMMC-7721 cells in 30 mg·L^－1 CXCL10+PD98059 group was decreased （P<0.05）. Conclusion CXCL10 can promote the proliferation and migration of the HCC SMMC-7721 cells， and its mechanism is mainly related to the up-regulation of the expressions of ERK/p-ERK/Cyclin D1 pathway proteins.

Objective To discuss the effect of cell division cyclin-dependentkinase like-1 （CDKL1） gene silencing on the proliferation， invasion， and metastasis of the breast cancer MCF-7 cells， and to clarify its possible mechanism. Methods The breast cancer MCF-7 cells were used as the research subjects， and the small interfering RNA （siRNA） technology was used to silence the expression of the CDKL1 gene. The intervention was performed by using 1 μmol·L^－1 phosphatase and tension homolog （PTEN） inhibitor BpV or 10 μmol·L^－1 protein kinase B（Akt） agonist SC79. The MCF-7 cells at logarithmic growth phase were divided into control group （MCF-7 cells without any treatment）， transfection control （siRNA-NC） group （MCF-7 cells transfected with siRNA-NC plasmid）， siRNA-CDKL1 group （MCF-7 cells transfected with siRNA-CDKL1 plasmid）， siRNA-CDKL1+BpV group （MCF-7 cells transfected with siRNA-CDKL1 and treated with 1 μmol·L^－1 PTEN inhibitor BpV for 2 h）， and siRNA-CDKL1+SC79 group （MCF-7 cells transfected with siRNA-CDKL1 and treated with 10 μmol·L^－1 Akt agonist SC79 for 2 h）. Real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to detect the expression levels of CDKL1 mRNA and protein in the breast cancer MCF-7 cells in various groups； CCK-8 assay was used to detect the proliferation activities of the breast cancer MCF-7 cells in various groups；EdU assay was used to detect the rates of EdU positive cells in the breast cancer MCF-7 cells in various groups；cell scratch healing experiment was used to detect the scratch healing rates of the breast cancer MCF-7 cells in various groups； Transwell chamber assay was used to detect the numbers of migration and invasion breast cancer MCF-7 cells in various groups；Western blotting method was used to detect the expression levels of matrix metalloproteinase-2 （MMP-2）， matrix metalloproteinase-9 （MMP-9）， PTEN， Akt， mammalian target of rapamycin （mTOR）， phosphorylated Akt （p-Akt）， and phosphorylated mTOR （p-mTOR） proteins in the breast cancer MCF-7 cells in various groups. Results Compared with siRNA-NC group， the expression levels of CDKL1 mRNA and protein in the breast cancer MCF-7 cells in siRNA-CDKL1 group were decreased （P<0.01）， the proliferation activity was decreased（P<0.01），the rate of EdU positive cells was decreased（P<0.01），and the numbers of migration and invasion cells were decreased （P<0.01）， the expression levels of MMP-2， MMP-9， p-Akt， and p-mTOR proteins in the cells were decreased （P<0.01）， and the expression level of PTEN protein was increased （P<0.01）. Compared with siRNA-CDKL1 group， the proliferation activities of the breast cancer MCF-7 cells in siRNA-CDKL1+BpV group and siRNA-CDKL1+SC79 group were increased （P<0.01），the rate of EdU-positive cells was increased（P<0.01）， the numbers of migration and invasion cells were increased（P<0.01）， the expression levels of MMP-2， MMP-9， p-Akt， and p-mTOR proteins in the cells were increased（P<0.01），and the expression level of PTEN protein in the cells was decreased （P<0.01）. Conclusion Silencing CDKL1 gene can inhibit the proliferation， invasion，and metastasis abilities of the breast cancer MCF-7 cells， and its mechanism may be related to the regulation of the PTEN/Akt/mTOR signaling pathway.

Objective To identify the key genes associated with the early diagnosis and poor prognosis of hepatitis B virus-associated hepatocellular carcinoma （HBV-HCC） by using the bioinformatics methods， and to elucidate the underlying molecular mechanism of occurence and development of HBV-HCC. Methods Gene Expression Omnibus （GEO） Database was used to retrieval “hepatitis B induced HCC”； the Gene Dataset GSE121248 was downloaded， the differentially expressed genes （DEGs） were screened by the “limma” Data Package in R Software， and the DEGs were enriched by using the “clusterProfiler” Data Package for Gene Ontology （GO） functional analysis and the Kyoto Genes and Genome Encyclopedia （KEGG） signaling pathway enrichment analysis； STRING Database and Cytoscape Software were used to establish the protein-protein interaction（PPI） network and the key genes were screened out.Gene Expression Profiling Interactive Analysis（GEPIA）， Kaplan Meier-Plotter，and Human Protein Atlas（HPA）Databases were used to verify the key genes and the expression levels of proteins；the infiltration of the immune cells was analyzed based on the “CIBERSORT” Data Package. Results A total of 574 DEGs were identified，including 173 up-regulated genes and 401 down-regulated genes. The GO functional enrichment analysis results showed that DEGs were mainly enriched in the biological processes such as small molecule metabolism， signal transduction， immune response， inflammatory response，and so on； the KEGG signaling pathway enrichment analysis results showed that the DEGs were mainly enriched in retinol metabolism， cytochrome P450 metabolic pathway of exogenous drugs， and chemical carcinogenesis，and so on. The PPI network results showed that cell division cycle 20（CDC20），cyclin dependent kinase 1（CDK1）， cyclin A2（CCNA2）， pindle checkpoint protein（BUB1B）， topoisomerase Ⅱ α（TOP2A）， discs large homolog associated related protein 5（DLGAP5）， abnormal spindle-like microcephaly associated protein（ASPM）， centrosomal protein 55（CEP55）， kinesin superfamily 11（KIF11），and kinesin superfamily 20A（KIF20A） were the key genes. The GEPIA Database analysis results showed that these above 10 key genes were highly expressed in the HCC patients. The Kaplan Meier survival curve showed that the overall survivals of the HCC patients with high expression of key genes were shorter than those of the HCC patients with low expression of key genes. Conclusion The genes related to cell cycle and viral oncogenesis （CDC20， CDK1， CCNA2， and BUB1B） are closely associated with the occurence and development and poor prognosis of the HBV-HCC patients， which may become the diagnostic markers and new targets for the treatment.

Objective： To analyze the serum metabolite compositions of the patients with esophageal squamous cell carcinoma （ESCC）， to screen out the potential differential metabolites， and to explore the relationship between the occurrence of ESCC and energy metabolism， and to provide the useful energy metabolites for the early diagnosis of ESCC. Methods The serum samples of the ESCC patients and healthy volunteers were collected and divided into ESCC group and control group， and there were 7 cases in each group. Liquid chromatography-tandem mass spectrometry （LC-MS/MS） method was used to detect the serum metabolites of the subjects in two groups；partial least squares discriminant analysis （PLS-DA） and orthogonal partial least squares discriminant analysis （OPLS-DA） models were used to identify the differences in serum metabolites of the subjects between ESCC group and control group， and they were classified according to the chemical classification of serum metabolites； targeted metabolomics analysis on serum energy metabolites was conducted based on the multi-response monitoring （MRM） mode to further identify the significantly differential energy metabolites and their pathways. Results A total of 266 metabolites were identified in the serum samples of the subjects in ESCC group and control group with the positive and negative ion modes， and there were 164 and 102 metabolites identified with the above two modes， respectively. The univariate statistical analysis results showed that there were significant differences in serum metabolites of the subjects between ESCC group and control group. Among the metabolites with chemical classification， lipids and lipid-like molecules accounted for the highest proportion （18.421%）. Three significantly different energy metabolites in ESCC group were found， and the expression levels of L-malic acid and isocitric acid were increased and the expression level of cyclic adenosine monophosphate （cAMP） was decreased. Conclusion There are differences in the expressions of serum energy metabolites between the ESCC patients and healthy controls， and the occurrence of ESCC is related to the energy metabolism.

Objective To discuss the relationship between microdeletions of 15 sequence tagged sites （STS） in azoospermia factor （AZF） region of the Y chromosome and male infertility （MI）， and to provide the evidence for the intervention of hereditary MI. Methods A total of 2 586 suspected MI patients were selected， and they were divided into four groups according to their ages，≤20 years old group（14 cases）， 21—30 years old group（988 cases）， 31—40 years old group（1 318 cases），and ≥41 years old group（266 cases）. The polymerase chain reaction （PCR） method was used to detect the 15 STS sequence fragments in the Y chromosome AZF region and the abnormal results were screened out. The microdeletion status of the Y chromosome was compared among the MI patients in various groups. Results Among 2 586 samples， 207 cases of Y chromosome abnormalities were found， accounting for 8.00% （207/2 586） of the total samples. The Y chromosome abnormalit rates of the somples in≤20 years old， 21—30 years old，31—40 years old， and ≥41 years old groups were 7.14% （1/14）， 8.10% （80/988），8.04% （106/1 318）， and 7.52% （20/266）， respectively； there were significant differences in the detetion rates of base sites and extension sites of the patients between various groups （χ²=10.836，P=0.013），and the deletion rate of base sites and extension sites of the patients in 21—30 years old group was higher than that in 31—40 years old group （P<0.05）. In the overall tested samples， 52 cases of base site fragment deletion were found， the abnormality rate was 2.01%， and there were significant differences in the abnormality rates of the patients between various groups （χ²=9.658， P=0.022）. The deletion rate of the AZFc segment accounted for 1.39% of all tested individuals， and the deletion rates of the patients in 21—30 years old group and 31—40 years old group were significantly higher than that in ≥41 years old group （P<0.05）.There were significant differences in the overall deletion rates of the patients between 21—30 years old and 31—40 years old groups（χ²=3.612，P=0.040）. There were no statistically significant differences in the deletion rates of the sY127， sY134 combined with sY105， sY121， sY1192， sY153， and sY160 sites of the patients between various groups （P>0.05），and there were no significant differences in the deletion rates of the sY254， sY255 combined with sY105， sY121， sY1192， sY153， and sY160 sites of the patients between various groups （P>0.05）. Conclusion The main cause of Y chromosome abnormalities of the males in reproductive age group in Northeast of Guangxi Zhuang Autonomous Region is microdeletion at the sY1192 and sY153 sites， and the sY1192 site microdeletion is the most prevalent. The detection rate of mutation at this site is increased with the increasing of age.

Objective To seek the prognostic markers for the gastric cancer （GC） patients， and to achieve the early diagnosis and treatment of GC， and to provide the evidence for improving the survival rate of the GC patients. Methods The clinical data and transcriptome data of 407 and 433 GC patients were downloaded from the The Cancer Genome Atlas（TCGA） Database and GSE84437 Dataset， and merged； the GC tissue samples were classified and typed based on the expression levels of cell death-related genes and the ConsensusClusterPlus Package，and were divided into type A（342 cases） and type B（465 cases）；the GC patients were divided into low expression group and high expression group according to the expression level of apolipoprotein D（APOD）；the survminer Data Package was used to compare the differences in prognosis of the patients with different subtypes；the ssGSEA Algorithm was used to compare the differences in immune cell infiltration of the patients with different subtypes； Lasso regression and Cox regression were used to construct the prognostic risk model for the GC patients； the clinical characteristics of model genes were filtered；the differential expression of APOD in adjacent normal tissue and GC tissue was analyzed by Public Databases；GSEA analysis was used to assess the Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment of APOD； the CIBERSORT Algorithm was used to evaluate the correlation between the content of immune cells and expression of APOD；the online website was used to analyze the drug sensitivity of APOD. Results There was statistically significant difference in prognosis of the patients with two subtypes of cell pyroptosis（P=0.002）； the levels of 22 kinds of immune cells of the patients with type A were higher than those with type B（P<0.01）； there were statistically significant differences in the percentages of two subtypes of cell pyroptosis of the patients with different ages （P<0.01） and N stages （P=0.04）. The Public Databases analysis results showed that the expression of APOD in tumor tissue of the GC patients was lower than that in adjacent normal tissue； the survival results showed that compared with low expression group，the patients in high expression group had poorer prognosis； the clinical correlation analysis results showed that there were significant differences in the APOD expression of the patients with different T stage （P<0.05）； the GSEA analysis results showed that high expression group enriched in the cell adhesion pathways， leukocyte endothelial migration pathways， and gap junction pathways； the immune infiltration analysis results showed that the contents of follicular helper T lymphocytes， CD4+ memory activated T lymphocytes， resting NK cells， and neutrophils in high expression group were lower than those in low expression group； APOD had positive correlations with CXC chemokine ligand 12（CXCL12）（r=0.500，P<0.01），transforming growth factor beta 1（TGFB1） （r=0.313，P<0.01），chemokine CC chemokine ligand 19（CCL19）（r=0.518，P<0.01），and CX3C chemokine receptor 1（CX3CR1）（r=0.444，P<0.01）；the drug sensitivity analysis results showed that there were positive correlations between APOD and AZD-7762， KW-2449， and TG-101348（0<r<0.3，P<0.05）. Conclusion Cell pyroptosis subtypes can effectively evaluate the prognosis of the GC patients； APOD can be regarded as the novel prognostic marker and therapeutic target for GC.

Objective To analyze the key genes and candidate pathways related to the occurrence and development of glioblastoma multiforme （GBM） by bioinformatics methods， and to explore the pathogenesis and therapeutic targets of GBM. Methods Gene Expression Datasets TCGA-GBM and GSE7696 were obtained from The Cancer Genome Atlas （TCGA） Database and Gene Expression Omnibus （GEO） Database. Deseq2 and limma R Data Packages were used to screen the differentially expressed genes （DEGs） in GBM tissue and adjacent normal tissue， and the Gene Ontology （GO） fuctional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were performed on the DEGs； the protein-protein interaction （PPI） network was analyzed by using the STRING Database； Cytoscape 3.9.1 Software was used to visualize the PPI network and perform the modular analysis. Results After the DEGs analysis of TCGA-GBM Transcription Data and Chip Dataset GSE7696， a total of 13 upregulated differentially expressed genes （UDEGs） and 77 downregulated differentially expressed genes （DDEGs） were obtained. The results of GO fuctional enrichment analysis showed that the DDEGs were mainly concentrated in the chloride channel activity， gamma-aminobutyric acid （GABA） receptor activity， GABA-gated chloride ion channel activity， GABA-A receptor activity， anterograde trans-synaptic signaling， chemical synaptic transmission and other biological processes. The KEGG signaling pathway were mainly concentrated in the GABA ergic synapse， neuroactive ligand-receptor interaction， neuro synapses containing serum， synaptic vesicle cycle and other signaling pathways. Two important gene modules were identified by PPI and module construction. The Cytoscape Software analysis results showed that solute carrier family 17 member 6（SLC17A6）， solute carrier family 1 member 2（SLC1A2），tachykinin precursor 1（TAC1），synaptotagmin 1（SYT1）， RNA binding fox-1 homolog 3（RBFOX3）， and gamma-aminobutyric acid type A receptor subunit gamma 2（GABRG2） were the key genes in PPI network. Conclusion SLC17A6，SLC1A2，TAC1，SYT1，RBFOX3，and GABRG2 genes may be involved in the occurrence and development of GBM，and the dysregulation of GABA ergic synaptic transmission related genes and pathway regulation network may be the main mechanism of pathogenesis of GBM.

Objective To discuss the expression levels of the cyclin-dependent kinase 5 regulatory subunit associated protein 1-like 1（CDKAL1） gene and its splice isoforms in peripheral blood lymphocytes of the patients with type 2 diabetes mellitus （T2DM）， and to clarify their clinical significances. Methods A total of 65 diabetes mellitus patients were enrolled， including 20 T2DM patients （T2DM group）， 23 diabetic nephropathy（DN） patients （DN group）， 22 diabetic retinopathy（DR）patients （DR group）， and 28 healthy examiners at the same term（healthy control group）. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of CDKAL1 gene and its two splice isoforms， CDKAL1-X1 and CDKAL1-X2， in peripheral blood lymphocytes of the subjects in various groups. The relationships between the clinical significances of their expressions and T2DM and its microvascular complications were analyzed. Results Compared with healthy control group， the expression levels of CDKAL1 gene （Z=4.705， P<0.01） and CDKAL1-X1 （Z=2.698， P=0.007） in peripheral blood lymphocytes of the patients in T2DM group were increased. Compared with healthy control group， the expression levels of CDKAL1 gene and CDKAL1-X1 in peripheral blood lymphocytes of the patients in DR and DN groups were increased（P<0.05）； the expression level of CDKAL1 gene in peripheral blood lymphocytes of the patients in DR group was higher than that in DN group （P<0.05）. Conclusion The high expressions of CDKAL1 gene and CDKAL1-X1 in peripheral blood lymphocytes of the T2DM patients may play an important role in the occurence and development of diabetic mellitus and its complications.

Objective： To analyze the effective chemical components and their action targets of epimedium in the treatment of hypothalamus-pituitary-adrenal gland/gonad/thyroid gland axis functional damage by using network pharmacology and molecular docking technology， and to preliminarily clarify its mechanism. Methods The effective chemical components and their corresponding target proteins of epimedium were screened by Traditional Chinese Medicine Systems Pharmacology （TCMSP） Database and Analysis Platform combined with the relevant literatures， and the Uniprot Database was used to standardize the target protein informations to get the corresponding standard gene names； the relevant target genes of hypothalamus-pituitary-adrenal gland/gonad/thyroid gland axis function damage were collected from the GeneCards Database and DisGeNET Database；the network of the relationship between the effective chemical components of the epimedium and their action targets was constructed by Cytascape 3.7.1 Software； the protein-protein interaction （PPI） network diagram of drug-disease of effective chemical components and target proteins of hypothalamus-pituitary-adrenal gland/gonad/thyroid gland axis function damage was constructed by STRING Database. Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genome （KEGG） signaling pathway enrichment analysis were performed by OmicShare Database；molecular simulation docking and visualization were done by Pymol Software and the Autodock Tools Software. Results A total of 27 effective chemical components and 217 corresponding action targets of epimedium， and 465 target genes of hypothalamus-pituitary-adrenal gland/gonad/thyroid gland axis function damage were screened out， of which 62 target genes were closely related to the treatment of hypothalamus-pituitary-adrenal gland/gonad/thyroid gland axis function damage by epimedium.Quercetin，luteolin，kaempferol， and icariin and so on were the key effective chemical components， and the key target proteins were Cysteine protease 3 containing cysteine（Caspase-3）， interleukin-6（IL-6）， oligosaccharides （FOS）， tumor necrosis factor（TNF）， hypoxia inducible factor-1 alpha （HIF1A）， vascular endothelial growth factor A（VEGFA）， interleukin-1β（IL-1β）， estrogen receptor 1 （ESR1），recombinant prostaglandin endoperoxide synthase 2（PTGS2）， matric metallopeptidase 9（MMP-9），and so on.The KEGG signaling pathway enrichment analysis results showed that the common targets were mainly concentrated in the pathways related to immune inflammation， hormone regulation， and energy metabolism. The molecular docking results showed that the potential active components of epimedium had a good affinity with IL-6， PTGS2， aldose reductase（AR）， and mitogen-activated protein kinase 1（MAPK1）targets. Conclusion Epimedium may play a potential therapeutic role in the treatment of hypothalamus-pituitary-adrenal gland/gonad/thyroid gland axis function damage in the aspects of anti-inflammatory immunity， hormone regulation， and energy metabolism through multiple targets and pathways.

Objective To discuss the risk factors for cognitive frailty in the elderly patients with chronic diseases，and to construct the corresponding risk model. Methods The general clinical data of 207 elderly patients with chronic diseases were collected.The FRAIL Scale and the Montreal Cognitive Assessment Questionnaire（MoCA） were used to detect the status of cognitive frailty of the patients， and the patients were divided into cognitive frailty group （n=75） and non-cognitive frailty group （n=132）. Univariate and binary Logistic regression analysis were used to identify the risk factors associated with cognitive frailty in the elderly patients. Results Among the 207 elderly patients， a total of 75 cases （36.23%） had cognitive frailty. There were significant differences in age， body mass index （BMI）， physical exercise， job category， social activity， night sleep time， daytime mental status， and number of chronic diseases of the patients between two groups（P<0.05）. After propensity score matching for age， the binary Logistic regression analysis results showed that low BMI， physically labour， night sleep time≤5 h， poor daytime mental status， and more than two chronic diseases were the risk factors for the cognitive frailty. The receiver operating characteristic curve （ROC） analysis results showed the area under curve（AUC） was 0.87 for the risk model constructed in this study to distinguish the patients’ cognitive frailty. Conclusion The risk model constructed based on the risk factors in this study has good predictive efficacy for cognitive frailty in the elderly patients，and its has potential clinical application prospects.

Objective To assess the bone thicknesses of the infrazygomatic crest in the adult patients with different vertical skeletal patterns of skeletal classⅠmalocclusion based on cone-beam computed tomography（CBCT），and to provide the basis for the choice of clinical treatment methods for the orthodontists. Methods A total of 90 adult patients with skeletal class I malocclusion were selected and divided into low angle group， average angle group，and high angle group， and there were 30 patients in each group. The buccal bone thickness of the patients at different vertical levels and proximal，distal and mesial levels were detected on the CBCT images and the effective bone volume was measured at buccal alveolar bone thickness ≥ 3 mm with the 50°，60°and 70° implantation angles. Results In different vertical sections， except for the proximal mesiobuccal root level of the maxillary first molar（U6_mb）， the buccal alveolar bone thickness was increased gradually from the top of the alveolar ridge to the root side（P<0.05）. At different proximal，distal and mesial levels， the buccal bone thickness of the patients was gradually increased from the proximal to the distal mesial direction in all levels except U6_mb（P<0.05）. The maximum buccal bone thickness of the patients was 11 mm from the top of the alveolar ridge at the proximal mesiobuccal root of the maxillary second molar（U7_mb）； the minimum buccal bone thickness was 7 mm from the top of the alveolar ridge at U6_mb.There was significant difference in the buccal bone thickness of the patients with different vertical bone facial types （P<0.05）， the buccal bone thickness of the patients in low angle group was higher than those in average angle group and high angle group （P<0.05），and the buccal bone thickness of the patients in average angle group was higher than that in high angle group （P<0.05）.Among the patients in low angle group， there were significant differences in bone thicknesses between the distal mesiobuccal root of the maxillary first molar and the proximal mesiobuccal root of the maxillary second molar （U6_db-U7_mb） at from the top of the alveolar crest was 7 mm，at U7_mb when the distance from the top of the alveolar crest was 7 mm and 9 mm， at U6_mb-U6_db when the distance from the top of the alveolar crest was 11 mm at three angles（P<0.05）.The bone thickness was ≥6 mm at U6_db-U7_mb， when the distance from the top of the alveolar crest was 7 mm at three angles， and when the distance from the top of the alveolar crest was 9 mm at 70° at U7_mb.Among the patients in average angle group， there were significant differences in the bone thicknesses between U6_db-U7_mb at 9 mm from the top of the alveolar crest， U7_mb at 9 mm and 11 mm from the top of the alveolar crest at three implantation angles，and the bone thickness ≥6 mm at U7_mb when the distance from the top of the alveolar crest at three angles was 7 mm and at U7_mb when the distance from the top of the alveolar crest was 9 mm at 70°（P<0.05）. Among the patients in high angle group， there were significant differences in bone thickness between U6_db-U7_mb 11 mm from the top of the alveolar ridge and U7_mb 11 mm from the top of the alveolar ridge at three implantation angles（P<0.05）. The thicknesses of bone implantation at different angles could be ranked as follows， 70°>50°>60° in all dimensions. Conclusion There are differences in the buccal bone thickness in the zygomatic crest area among the adult patients with different vertical facial bone patterns. The patients in low angle group have more options for the implantation sites， while the patients in high angle group are more prone to maxillary sinus perforation.Therefore，when placing dental implants and designing orthodontic treatment plans in this area，the vertical facial bone pattern should be taken into consideration.

Objective To observe the changes of sagittal plane balances of cervical spine of the patients underwent posterior cervical single open-door expansive laminoplasty， and to provide the imaging evidence for the postoperative rehabilitation training of the patients. Methods A total of 32 patients who underwent posterior cervical single open-door expansive laminoplasty were selected. According to the median value（15.75 mm ）of sagittal vertical axis （SVA） distance before operation， the patients were divided into low SVA group and high SVA group，and there were 16 patients in each group. The imaging and clinical data of the patients in two groups before operation and at the last follow-up were retrospectively analyzed. The SVA value， cervical lordosis angle （Cobb angle）， and T1 tilt angle （T1s） of the patients at the X-ray lateral view of the cervical spine were detected before operation and at the last follow-up. The postoperative Japanese Orthopaedic Association （JOA） score， neck disability index （NDI） store，and satisfaction score of the patients in two groups were analyzed. Results Compared with before operation， the NDI score of the patients in high SVA group after operation was decreased （P<0.01）， the JOA score was increased （P<0.01）. Compared with before operation， the NDI score of the patients in low SVA group was decreased （P<0.01）， the JOA score was increased （P<0.01）， and the SVA value was increased （P<0.01）， while there were no significant differences in the Cobb angle and T1s （P>0.05）. There was no statistically significant difference in the occurrence of axial symptoms of the patients between low SVA group and high SVA group（P>0.05）. Conclusion In the follow-up of at least 2 years after operation， posterior cervical single open-door expansive laminoplasty has the certain effect on the sagittal plane balance of cervical spine of the patients. The main manifestations are tendency of cervical lordosis and anterior shift of the center of gravity， but the overall stability can still be maintained. The patients with high SVA have a higher incidence of axial symptoms after operation.

Objective：To discuss the application of the modified needle-through-needle puncture technique in unilateral total hip arthroplasty in the elderly patients，and to clarify its anesthetic effect and feasibility in the single isospecific gravity spinal anesthesia guided by ultrasound. Methods Sixty elderly patients underwent elective unilateral hip arthroplasty with single isobaric gravity spinal anesthesia were selected. The puncture was performed in the patients with the affected side upper in the lateral position. Sixty patients were divided into tradition group and modification group， and there were 30 patients in each group. After the ultrasonic localization of spinous process space， the single subarachnoid puncture through median approach was performed in all the patients， and the successful puncture was determined by the outflow or drawn-out of cerebrospinal fluid in the spinal anesthesia needle.The patients in tradition group were treated with the traditional needle-through-needle puncture technique （a 16 G epidural puncture needle was used to guide the puncture，and a 25 G pencil-point side-hole spinal needle was applied）；the patients in modification group were treated with the modified needle-through-needle puncture technique （a 20 G injection needle was used to break the skin directly for guidance， and a 25 G pencil-point side-hole spinal needle was applied）. The once-puncture success rates，twice-puncture success rates，first-skin-attempt puncture success rates， and twice-skin-attempt puncture success rates， puncture time， patients’ satisfaction rates，onset-time of anesthesia，assessment of anesthesia block， fixation-time of anesthesia， satisfaction of surgeon for muscle relaxation， operation time， residence time in PACU， immediate adverse reactions during anesthesia， and adverse reactions within 48 h after operation of the patients in two groups were compared. Results All the patients had successful punctures. Compared with tradition group， the once-puncture success rate， twice-puncture success rate， first-skin-attempt puncture success rate， and twice-skin-attempt success rate of the patients in modification group were increased （P<0.01），the puncture time was shorter （P<0.01）， the patients’ satisfaction rate was increased （P<0.01）， the onset-time of anesthesia was shortened（P<0.01），the fixation-time of anesthesia block was shortened（P<0.01）， the satisfaction rate of the surgeon was increased （P<0.01），the mean arterial pressure（MAP） and heart rater（HR） of the patients 10 min after anesthsia and 10 min after operation were increased（P<0.05）.Compared with tradiation group， the incidence of immediate adverse reactions during anesthesia， and the incidence of adverse reactions within 48 h after operation of the patients in modification group had no significant differences （P>0.05）. Conclusion In the elderly patients underwent unilateral total hip arthroplasty， the single isospecific gravity spinal anesthesia under non-visual conditions by the modified needle-through-needle puncture technique can significantly shorten the number of punctures and puncture time，and has higher puncture success rate.

Objective To observe the efficacy of dupilumab in the treatment of refractory atopic dermatitis （AD），and to provide the reference for the treatment of such patients. Methods The clinical data and follow-up results of two patients with refractory severe AD treated with dupilumab were collected；combined with the literature review， the efficacy and safety of dupilumab in the treatment of refractory AD were analyzed. Results Patient 1 was a 30-year-old female with dense， infiltrated papules on the back of both hands， resembling lichenified skin changes， scattered erythema， and papules with itching on the limbs and trunk for 25 years. Patient 1 also had a history of allergic rhinitis for 5 years. The diagnosis was adults severe AD. After 16 weeks of treatment with dupilumab， the skin lesions were completely regressed， and there were no relapse during the follow-up period. The symptoms of allergic rhinitis was improved significantly， and oral medication was no longer needed. Patient 2 was a 59-year-old female with general dark red erythema， papules， and nodules accompanied by itching for over 10 years. Patient 2 had been hospitalized many times and had previously received treatment with cyclosporine， azathioprine， and steroids with unsatisfactory results. The diagnosis was adults severe AD. Up to the present， the skin lesions had essentially regressed after treatment with dupilumab. Conclusion Dupilumab has good curative effect and safety on refractory AD，and it can improve the life quality， and controll the charicteristics of AD complicated with other allergic diseases driven by type 2 inflammation.

Objective To observe the clinical manifestations and treatment outcomes of 40 patients with Fournier’s gangrene （FG）， and to explore the application prospects of modified negative pressure drainage and irrigation technique in the treatment of FG. Methods A retrospective analysis on the clinical data of 40 FG patients collected in our department was conducted. The patients were treated with surgical debridement combined with negative pressure irrigation technique. The etiology， complications， clinical manifestations， pathological examination results， laboratory examination results， wound bacterial culture， and postoperative hospital stay of the patients were recorded； and the wound healing and long-term follow-up outcomes of the patients were observed. Results Among these 40 patients， there were 35 males and 5 females， whose age were from 22 to 81 years old. There were 28 cases with perianal infection， 11 cases of diabetes， 5 cases of trauma， 1 case of pressure sore due to paraplegia， and 6 cases with no obvious cause. The pathological examination results showed the chronic suppurative inflammation with abscess formation. The surgical debridement was performed， and the wound treatment was carried out by covering the wound with negative pressure and modified drainage and irrigation technique based on the wound condition. All the patients were cured and discharged without recurrence. The average hospital stay was 24.65 d. Among the 25 patients with perianal abscess， no obvious stenosis occurred after surgery； 8 patients with involvement of the scrotum recovered well with normal appearance and function； 7 patients with skin grafting had good growth of the grafts； 2 patients with flap repair had good blood supply to the flaps without redness or rupture； 8 patients with involvement of the lower limbs had good recovery of motor and sensory function without obvious disabilities or hypertrophic scars. Conclusion The use of surgical debridement combined with negative pressure drainage and irrigation technique in the treatment of FG is beneficial for the observation of the changes in the wound， reducing dressing changes， shortening the hospital stay， improving the cure rate， and is worthy of clinical promotion.

Objective To compare the clinical efficacies of transurethral plasmakinetic enucleation of prostate （PKEP） and transurethral plasmakinetic resection of prostate （PKRP） in the treatment of benign prostatic hyperplasia （BPH）， and provide the evidence for the treatment options for BPH. Methods The clincal data of 60 patients with lower urinary tract symptoms （LUTS） and initial diagnosis as BPH were collected. With the patients’ informed consent， they were divided into PKEP group （treated with PKEP） and PKRP group（treated with PKRP）， and there were 30 cases in each group. The age， body weight， body mass index （BMI）， total prostate-specific antigen （tPSA） level， hematocrit （HCT）， hemoglobin （Hb） level，preoperative and postoperative sodium ion level， prostate volume，degree of midlobe median lobe protrusion， occurrence of complications （hypertension， diabetes， and respiratory diseases）， percentage of oral 5α-reductase inhibitor using， operation time，amount of intraoperative blood， resected tissue volume， resection rate，resection efficiency，total hospital stay after operation，indwelling catheterization time， bladder irrigation duration， sodium ion level，Quality Of Life （QOL） score， and International Prostate Symptom Score （IPSS） before and after operation of the patients in two groups were observed and compared.The incidences of adverse reactions of the patients in two groups after operation were also compared. Results There were no significant differences in age， body weight， BMI，tPSA level， preoperative HCT， preoperative Hb level，sodium ion level before operation， prostate volume， and degree of midlobe protrusion of the patients between two groups （P>0.05）. There were no statistically significant differences in the occurrences of complications （hypertension， diabetes， and respiratory diseases） and the percentage of oral 5α-reductase inhibitor using of the patients between two groups （P>0.05）； the amount of blood during operation， resected tissue volume， resection rate， and resection efficiency of the patients in PKRP group were higher than those in PKEP group （P<0.05）； there were no significant differences in the total hospital stay，indwelling catheterization time， and bladder irrigation duration after operation of the patients between two groups （P>0.05）； there were no significant differences in the sodium ion levels， QOL scores， and IPSS of the patients between two groups before and after operation（P>0.05）； there were significant differences in the QOL scores and IPSS of the patients in each group before and after operation （P<0.05）.There was one case of urinary incontinence in PKEP group and two cases of urinary incontinence in PKRP group， without other operation-related complications. Conclusion Both two methods have similar efficacy and safety， and can achieve the satisfactory surgical outcomes.The patients undergoing PKEP have less amount of blood during operation， higher resected tissue volume，the higher resection efficiency， and higher resection rate， which is a better way to treat BPH.

Objective To observe the therapeutic effect of 3D-printed titanium alloy sternum in the postoperative reconstruction of sternum in the patients with failed conventional titanium plate stermal reconstruction， and to provide the evidence for the treatment of such disease. Methods The clinical data of one patient with malignant sternum tumor were collected. The patient’s operation process was observed， and the recovery of the patient’s physical function was followed up. The relevant literatures were reviewed and summarized. Results One patient， a 59-year-old male， presented with sternum pain without obvious cause and received the chest computed tomography（CT） examination， which indicated a space-occupying lesion in the sternum. The patient was admitted with the diagnosis of “sternal tumor” and underwent partial sternectomy and conventional titanium plate placement in 2017. Subsequently， the patient experienced severe restriction of limb movement， titanium plate fracture， and deformation 1 year after operation. In 2019， 3D-printed titanium alloy sternum implantation was performed to repair the sternum defect and reconstruct the thoracic cage.Compared with before operation， the patient’s limb movement gradually recovered， ranging from passive to active activities， and the normal range of motion of the shoulder joint was gradually improved significantly after operation. In 2020， due to the thin subcutaneous fat and stable formation of fibrous connective tissue， the planned operation was performed to remove the 3D-printed titanium alloy sternal rib clavicle plate. The postoperative recovery of the patient was smooth， and the upper limb activity was normal and the quality of life was significantly improved.The prognosis was good. Conclusion The application of 3D-printed titanium alloy sternum enables precise personalized treatment， which is superior to the conventional titanium plate， and it can ensure the anatomical restoration and reduce the occurrence of complications in the patients.

Objective To construct a pig-mouse liver cell chimeric model，and to discuss the relevant conditions for the transplantation and regeneration of the porcine hepatocytes without T lymphocyte， B lymphocyte， and NK lymphocyte mediated rejection， and to evaluate the settlement， proliferation， and secretion of functional factors of the porcine hepatocytes in xenograft receptors， and to provide the basis for improving the reconstruction efficiency of the porcine hepatocytes in pig derived mouse models. Methods The porcine hepatocytes were transplanted into the Fah^－/－Rag2^－/－ IL2Rg ^－/－ （FRG） mice by using intrasplenic injection method to construct the transplantation model. Sixteen FRG mice were randomly divided into control group， blank control group， experimental group， and green fluorescence protein（GFP） gene experimental group， and there were four mice in each group.The administration of nitisinone 2-2-nitro-4-（trifluoromethyl）cyclohexane-1，3-dione（NTBC） of the mice in blank control group was stopped，and the body weight was monitored； the mice in control group were injected with the porcine hepatocytes， the administration of NTBC was stopped one week before transplantation， and the body weight was monitored after transplantation. When the body weight was decreased by 20%， the NTBC was administered for 3 d. The mice in experimental group were injected with the porcine hepatocytes， and the administration of NTBC was stopped one week before transplantation； the body weight was monitored after transplantation， and when the body weight was decreased by 20%， the NTBC was administered for 3 d. The mice in GFP gene experimental group were injected with the porcine hepatocytes carrying the GFP gene， the administration of NTBC was stopped one week before transplantation， and the body weight was monitored after transplantation； when the body weight was decreased by 20%， the NTBC was administered for 3 d. The body weights and survival status of the mice in various groups were observed. The levels of alanine aminotransferase （ALT） in serum of the mice in various groups were detected；the expressions of CD19+， CD3+， and NK1.1+cells in peripheral blood of the mice in various groups were detected by flow cytometry；the expression levels of porcine albumin in serum of the mice in various groups were detected by ELISA assay； the regeneration efficiencies of the porcine hepatocytes of the mice in various groups were detected by immunohistochemistry. Results Compared with method of collagenase digestion after shearing， the digestion process of the two-step perfusion method was milder and had less damage to cells， and the cell activity achieved 92%. After stopping the administration of NTBC， the body weight of the mice in control group was gradually decreased and the vitality was gradually decreased. Death began to occur in the mice on the 28th day after continuous stopping the administration.The irreversible cellular damage in liver tissue of the mice could be seen such as liver cell enlargement， cytoplasmic looseness and transparency， and nuclear lysis. The ALT levels in serum of the mice in blank control group was gradually increased after stopping the administration. After re-administration of NTBC， the body weight of the mice in control group was gradually returned to the normal levels. The flow cytometry results showed that the CD19+， CD3+， and NK1.1+cells in peripheral blood of the mice could not be seen after stopping the administration of NTBC. The immunohistochemistry staining results showed that no dark Fah+positive cells were found in liver tissue of the mice in experimental group， while the dark Fah+positive cells were observed in liver tissue of the transplanted mice； the regeneration efficiency of porcine hepatocytes was （11 ± 4）%. The regeneration efficiency of porcine hepatocytes of the mice in GFP gene experimental group was （10 ± 2）% when stimulated by three color lasers in the same field of view. Conclusion The pig derived FRG mouse liver chimeric models are successfully constructed， and the mouse models for long-term stable expansion of the porcine hepatocytes are established.

Objective The human-derived antimicrobial peptide（AMPs） histatin 5 （Hst5） and LL-37 were used as the parents to design a novel heterozygous peptide H5LL， and to construct a recombinant lactic acid bacteria expression vector by genetic engineering technology to achieve the efficient and safe expression of the AMPs. Methods Bioinformatics method was used to predict the physicochemical parameters， hydrophilicity/hydrophobicity， shear sites， phosphorylation sites， signal peptides and transmembrane regions， subcellular localization， and secondary structure of H5LL； the target sequence and the empty plasmid pMG36e were doubly digested by HindⅢ and KpnⅠ， then the lactic acid bacteria recombinant expression vector pMG36e-H5LL was constructed and cloned to obtain the recombinant plasmid containing the target genes. Results H5LL had a high possibility of being an AMPs， contained 36 amino acids， the relative molecular weight was 4 625 380，the total charge number was +10， which had hydrophilicity； there were 3 phosphorylation sites and there was no glycosylation site； H5LL belonged to intramembrane protein， without transmembrane region and signal peptide；the subcellular localization prediction results showed that the possibility of mitochondrial targeting peptide was 0.333. The α-helix junction and β-turned angle accounted for 52.78% and 22.22% in the secondary structure， and the number of α-helix was the highest in the tertiary structure. The PCR electrophoresis results of recombinant plasmid contained target gene showed a single specific band at 138 bp； the recombinant plasmid double digestion electrophoresis results showed the clear bands at 136 and 3 500 bp. Conclusion The novel heterogeneous peptide H5LL is designed and synthesized with high stability， antibacterial activity and low toxicity；the recombinant lactic acid bacteria expression vector pMG36e-H5LL is successfully constructed.