Objective To establish a rat model of peritoneal bacterial infection after liver transplantation.Methods To construct a dark Agouti rat-to-Lewis(DA-to-LEW) rat model of liver transplantation.Peritoneal bacterial infection in the rats was induced by intraperitoneal injection of bacterial suspension.The liver function,blood gas,blood cell count and other indicators of the rat models were detected.Results There was a high mortality rate in rats with bacterial injection at day 5 after liver transplantation,therefore unfavorable for the following study.It waft better to inject the bacteria in an amount of 5×10~5 cfu/mL at day 3 after liver transplantation.The cumulative 7-day survival rate of those rats after infection reached up to 37.5%.The infection became increasingly severe,the general conditions were worsening,the rectal temperature was rising,the WBC count was increased,the pH was decreased,liver dysfunction was progressively increased,and metabolic acidosis occurred in the rats.Liver parenchymal damage was more pronounced than that of bile ductal injuries,and the rats died one after another at about 5 days after infection.Pathological examination of multiple organs showed that the main cause of death of the rats was liver damage,without accompanying lung and kidney damages.Conclusion The results of this study suggest that it is a successful method to establish a rat model of peritoneal bacterial infection after liver transplantation,and this model can be used in related experimental researches.

Objective To investigate the changes of immune state and the impact on immunological rejection elicited by abdominal cavity bacterial infection after DA-Lewis rat liver transplantation.Methods Orthotopic liver transplant model was established by modified Kamada two-cuff technique.The animals were divided randomly into Group 1,isotonic Na chloride injected into abdominal cavity 3 days after operation;Group 2,mixed Bacillus coli liquid injected instead of saline;Group 3,immunosuppressive drug CsA administered routinely after operation(3 mg?kg-1?d-1).All the animals were sacrificed 7 days after infection.The blood and graft samples were collected for cell-subpopulation,mixed lymphocyte culture,IL-4,IFN-? mRNA detection and histological evaluation.Results Seven days after infection,the lympholeukocyte population,CD4/CD8(G1=1.753?0.181,G2=1.384?0.073,G3=0.997?0.025)and lympholeukocyte function(SI:G1=67.59?3.40,G2=37.14?0.90,G3=15.87?0.51)declined in Group 2 as compared with other groups and cellular differentiation drifted to Th2.There was significant difference between Group 2 and Group 1 or 3.Conclusion Abdominal cavity bacterial infection after rat liver transplantation will promote the differentiation of T cells into Th2,down-regulate CD4/CD8 ratio and immune function of lymphocytes and accordingly alleviate partly the acute rejection following liver transplantation.

Objective To observe the clinical efficacy of needle-knife therapy in treating allergic rhinitis. Method Sixty allergic rhinitis patients were randomized into a treatment group and a control group, 30 cases in each group. The treatment group was intervened by needle-knife therapy; the control group was given Azelastine hydrochloride nasal spray plus oral administration of Desloratadine, both twice a day. The intervention lasted for 4 weeks in both groups. The Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ) and symptoms scores were observed before and after the treatment, and the clinical efficacies were compared between the two groups. Result The markedly effective rate was 90.0% in the treatment group versus 66.7% in the control group, and the between-group difference was statistically significant (P<0.05). The RQLQ and symptoms scores were significantly changed after the intervention in both groups (P<0.05). After the treatment, the RQLQ and symptoms scores in the treatment group were significantly different from those in the control group (P<0.05). Conclusion Needle-knife therapy is safe and effective in treating allergic rhinitis.

Objective By detecting the variation of long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) DNA methylation in preeclampsia-like mouse models generated by different ways, to explore the roles of multifactor and multiple pathways in preeclampsia pathogenesis on molecular basis. Methods Established preeclampsia-like mouse models in different ways and divided into groups as follows: (1) Nw-nitro-L-arginine-methyl ester (L-NAME) group: wild-type pregnant mouse received subcutaneous injection of L-NAME;(2) lipopolysaccharide (LPS) group:wild-type pregnant mouse received intraperitoneal injection of LPS; (3) apolipoprotein C-Ⅲ (ApoC3) group: ApoC3 transgenic pregnant mouse with dysregulated lipid metabolism received subcutaneous injection of L-NAME;(4)β2 glycoprotein I (β-2GPI) group:wild-type pregnant mouse received subcutaneous injection ofβ-2GPI. According to the first injection time (on day 3, 11, 16 respectively), the L-NAME, LPS and ApoC3 groups were further subdivided into:pre-implantation (PI) experimental stage, early gestation (EG) experimental stage, and late gestation (LG) experimental stage.β-2GPI group was only injected before implantation. LCHAD gene methylation levels in placental were detected in different experimental stage. Normal saline control groups were set within wild-type and ApoC3 transgenic pregnant mice simultaneously. Results (1) CG sites in LCHAD DNA:45 CG sites were detected in the range of 728 bp before LCHAD gene transcription start site, the 5, 12, 13, 14, 15, 16, 19, 24, 25, 27, 28, 29, 30, 31, 32, 34, 35, 43 CG sites were complex sites which contained two or more CG sequences, others were single site which contained one CG sequence. The 3, 5, 6, 11, 13, 14, 18, 28 sites in L-NAME, LPS, ApoC3 and β-2GPI groups showed different high levels of methylation; the 16, 25, 31, 42, 44 sites showed different low levels of methylation; other 32 sites were unmethylated. (2) Comparison of LCHAD gene methylation between different groups:the methylation levels of LCAHD gene at 3, 11, 13, 14, 18 sites in L-NAME, LPS, ApoC3 andβ-2GPI groups were significantly higher than those in the normal saline control group (P<0.05); and the methylation levels of 42, 44 sites in these groups were significantly lower than those in the normal saline control group (P<0.05). (3) Methylation of LCHAD gene at the same site between different experimental stages: ① The 3, 11, 18 sites of EG experimental stage was significantly lower than PI and LG experimental stage in L-NAME group (P<0.05);the 3, 11, 18 sites of PI experimental stage was significantly lower than EG and LG experimental stage in LPS group (P<0.05);these sites of PI experimental stage was significantly higher than EG and LG experimental stages in ApoC3 group (P<0.05).②The methylation of site 5 in L-NAME and LPS groups were significantly higher than that of the normal saline control group (P<0.05), and the LG experimental stages were significantly higher than other stages, but in ApoC3 group , only PI and EG stages were significantly higher than the normal saline control group (P<0.05).③At site 6 in L-NAME group which showed high methylation level was significantly higher than the same site in other groups which showed low methylation level (P<0.05).④At 13, 14 sites, earlier preeclampsia onset caused a lower methylation level in L-NAME group, but PI experimental stage was significantly higher than EG and LG experimental stages in LPS group (P<0.05), EG experimental stage was significantly higher than PI and LG experimental stages in ApoC3 group (P<0.05). ⑤ At site 28, earlier preeclampsia onset caused a higher methylation level in L-NAME group, but PI experimental stage was significantly lower than EG and LG experimental stages in LPS group (P<0.05), EG experimental stage was significantly higher than PI and LG experimental stages in ApoC3 group (P<0.05).⑥The 16, 25, 31 sites in ApoC3 group were significantly higher than other groups (P<0.05). ⑦ At site 42 in β-2GPI group was unmethylated, but it in other groups showed low methylation level, the methylation level of site 42 inβ-2GPI group was significantly lower than that in other groups (P<0.05). Conclusions The methylation of 6 and 42 CG sites may be related to LCHAD gene expression in placenta of L-NAME and β-2GPI induced preeclampsia-like models respectively;LCHAD gene expression and DNA methylation may not have obviouscorrelation in LPS and ApoC3 induced preeclampsia-like models. Differences exist in LCHAD DNA methylation in preeclampsia-like models generated by different ways, revealed a molecular basis to expand our understanding of the multi-factorial pathogenesis of preeclampsia.

Objective To investigate the effect of autophagy on the proliferation and migration of cervi-cal cancer cells ,as well as the underlining mechanisms .Methods Rapamycin was used to induce the autophagy in HeLa cells,formation of autophagosomes was observed by staining with acridine orange under fluorescence mi -croscope.Western blot was used to detect the expression of LC 3 and PI3K/Akt/mTOR in HeLa cells.The LC3 plasmid was transfected into HeLa cells .The distribution of LC3 in cells and the expression of LC3 was identified by fluorescence microscope and Western blot ,respectively.The autophagy was inhibited with 3-methyladenine(3-MA )in HeLa cells.The cell proliferation was monitored by RTCA real -time instrument.Transwell chamber was carried out to assess cell migration .Results After 6h of rapamycin treatment ,the expression of LC3B was in-creased in HeLa cells ( P<0 .05 ) .The proliferative and migration ability were weakened compared to wild type HeLa cells(P<0.05).The same results in the presence of 3-MA.The expression of PI3K/AKT/mTOR path-way proteins were activated by rapamycin treatment and LC 3 overexpression(P<0.05).Conclusion Autophagy can suppress the proliferation and migration in cervical cancer cells ,which may relate to PI3K/Akt/mTOR path-way.

Objective To study the value of three dimensional contrast enhanced subtraction MRA(3D CES MRA)in the diagnosis of body and lower extremity blood vessel disease Methods Eighteen cases were studied by 3D ECS MRA and proved by operationResults 3D CES MRA images were of diagnostic quality without ghost for all 18 patients.3D CES MRA clearly showed normal vascular anatomy and various disorders.According to surgery,the sensitivity and specificity of 3D CES MRA in the diagnosis of body and lower extremity blood vessel diseases were both 100%.Conclusion 3D CES MRA is a new technique in diagnosing of body and lower extremity blood vessel diseases.It is proved to be a simple,noninvasive and commercial method in diagnosing blood vessel diseases.

AIM: To investigate the effects of Lipopolysaccharide(LPS) and interleukin 1 receptor antagonist(IL-1ra) on mesangial cells proliferation and nitric oxide synthesis. METHODS: Glomerular mesangial cells from SD rats were cultured. The first and second passages of cultured cells were used for the experiment. LPS and LPS plus IL-1ra were added in cell cultures, respectively. By using chemical method the nitrite in supernatants was measured,3H-TdR incorporation was determined to evaluate the GMC proliferation. Northern and slot hybridizations were performed to detect the expression of iNOS mRNA. RESULTS: There were expression of iNOS mRNA, more production of nitrite(0.64?0.25 vs 0.12?0.06 nmol/104 cell) in supernatants and GMC proliferation(3735?1177.9 vs 1785?280.6) in LPS group compared to the control. While compared with LPS group, in LPS+IL-1ra GMC group, expression of iNOS mRNA decreased by 40%, nitrite increased(3.28?0.33 nmol/104 cell), proliferation of GMC decreased (818?77.27). CONCLUSION: LPS could activate the GMC to express iNOS mRNA and produce more nitrite. IL-1ra could partially inhibit the effects of LPS on the expression of iNOS mRNA in GMC, but not nitrite. There is no synchronous correlation between NO production and GMC proliferation.

Objective To study the effect of Chinese herbs on human scalp hair growth in vitro. Methods Organ culture of human scalp hair follicles was used to study the effects of extracts of Crataegus cuneata, Ligustrum lucidum, Polyporus umbelatus and Bletilla striata mixture and ginseng saponin on hair growth. Results Low doses of mixture extracts(1.28 ?g/mL and 6.4 ?g/mL) markedly enhanced the hair growth and lengthened the period of hair growth, while high doses of mixture extracts(4 mg/mL and 20 mg/mL) sharply inhibited hair growth and shortened the period of hair growth. High doses of ginseng saponin (40 ?g/mL and 200 ?g/mL) also had an inhibiting effect on hair growth. Conclusion This observation suggests that extracts of Crataegus cuneata, Ligustrum lucidum, Polyporus umbelatus and Bletilla striata mixture can promote hair growth in vitro.

Objective To analyze type IV collagen a5 chain mRNA and to study the relationship between genotype and phenotype in X-linked Alport syndrome. Methods Total RNA was isolated from the cultured skin fibroblasts of 21 unrelated Chinese X-linked Alport syndrome patients (17 males and 4 females),then ?5 chain (Ⅳ) mRNA was analyzed by using reverse-transcription-polymerase reaction (RT-PCR) and direct sequencing. Meanwhile,by using PCR and direct sequencing,detection of COL4A5 gene mutations at genomic DNA level was carried out. Results Abnormal ?5 chain IV cDNA was detected in 21 X-linked Alport syndrome patients,and seventeen mutations detected in this study were novel mutations. In 15/21 of patients,identical COL4A5 mutations were detected both at mRNA level and genomic DNA level,and in 6/21 of patients with splicing-site mutations,changes in transcript structure differed from changes in genomic DNA level. 16/21 of patients belonged to X-linked juvenile kindreds,and 2/21 of patients to adult kindreds. Conclusion Different type of mutations in COL4A5 can lead to the severe form of X-linked Alport syndrome,and mRNA-based procedures can both directly detect mutations in the coding sequences,as well as changes in transcript level or structure,and can identify some abnormalities that would otherwise have been missed by DNA-based procedures.

Objective To explore the effect of suledexide on renal injury and podocalyxin expression of podocytes in STZ diabetic desoxycorticosterone acetate (DOCA)-hypertensive rats. Methods Wistar rats were subjected to subcutaneous injection of streptozotocin(STZ), followed by uninephrectomy and subcutaneous administration of DOCA. Diabetic and hypertensive rats were randomly allocated to treatment with sulodexide or a combination of sulodexide and telmisartan for 8 weeks. Blood pressure (BP), 24-hour urinary albumin were measured every 2 weeks. Blood and urinary samples were collected to detect biochemical indexes of plasma and urinary β-acetyl-β-D-glucosaminidase (NAG) at the end of the study. Immunohistochemistry (IHC), RT-PCR and Western blot were performed to examine the expression and distribution of podocalyxin. Results STZ +DOCA-treated rats progressively developed hypertension, albuminuria and hyperglycemia. Hyperlipidemia and hypoinsulinemia were found in diabetic and hypertensive rats compared with controls. Albuminuia was significantly reduced in sulodexide group at week 8 and sulodexide plus telmisartan group at week 6 and week 8. Blood pressure decreased in sulodexide plus telmisartan group. No significant effects on lipid and glucose metabolism were observed in all treated groups. Histopathological index increased in STZ+DOCA-treated rats, but was significantly lower in sulodexide group as well as sulodexide plus telmisartan group. The number of podocytes on glomerular cross-section of the four groups were comparable. Segmental loss and down-regulation of podocalyxin were detected in STZ+DOCA-treated rats, which were greatly attenuated by sulodexinde, meanwhile, combination treatment preserved more podocalyxin expression in glomeruli than sulodexide monotherapy. Conclusion Sulodexide effectively reduces albuminuria, prevents loss of podocyte podocalyxin and alleviates renal damage in STZ diabetic DOCA-hypertensive rats.