[Objective]To investigate the mechanism of Qingre Yiqi Tongluo(QRYQTL) fang in treating the diabetic nephropathy. [Methods]60 Sprague-Dawleg rats were divided into normal control group(N), Diabetic nephropathy model group(M) (model rats were prepared by intraperitoneal injection with STZ after unilateral nephrectomy), then the model rats were assigned into model control group, QRYQTL fang treatment group(Q) and Irbesartan treatment group(I). After 12 weeks of the treatment, the contents of superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-Px) and malondialdehyde (MDA) in renal tissue were examined by enzyme linked immunoassay(ELISA); the glomerular area(GA), mesangium area(MA),MA/GA, mean width of the podocyte foot processes(FPW) ,foot process fusion rate(FPFR) and average thickness of glomerular basement membrane(GBMT) were measured. [Results]Compared with N group, the contents of MDA in kidney homogenate and the GA,MA, FPW,FPFR,GBMT were markedly increased (P<0.01or P<0.05),the contents of SOD,CAT, GSH-Px and MA/GA were significantly decreased( P<0.01or P<0.05). QRYQTL fang reduced the level of MDA and decreased the GA, FPW, FPFR and GBMT(P<0.01or P<0.05). Compared with I group ,the QRYQTL fang was superior to Irbesartan in reducing the MDA, FPFR as wel as increasing the SOD,CAT(P<0.05).[ Conclusions]QRYQTL fang has a protective effect on the experimental diabec-tic nephropathy rats. The mechanism may be related to inhibiting oxidative stress, improving the body ’s antioxidant ability and al eviating the glomerular morphologic injury.

[Objective]To study the effect of Qingre Yiqi Tongluo(QRYQTL) preseription on proteinria in diabetic nephropathy(DN) rats by means of ob-servation on the expressions of fibroblast-specific protein 1(FSP1) and P-cadherin in in glomerulus in rats.[Methods]60 Sprague-Dawleg rats were divided into normal control group(N), Diabetic nephropathy model group(M), QRYQTL preseription treatment group(Q) and Irbesartan treatment group(I). DN model rats were prepared by intraperitoneal injection with STZ after unilateral nephrectomy. The rats were sacrificed after 12 weeks of the treatment. The creatinine clearance(Ccr),blood urea nitrogen(BUN) and 24h urinary protein(UPro) were tested. Immunohistochemical staining was performed to explore the expressions of P-cadherin and fsp1 in glomerulus.[Results] ①Compared with M group, the levels of 24-hour upro in Q group were significantly de-creased(P<0.01).The levels of Ccr and BUN in Q group were both lower than those of M group( P<0.05). ② Compared with M group, the expressions of P-cadherin in glomerulus in Q group were increased( P<0.01) and the expressions of FSP1 were obviously decreased( P<0.01). The expressions of FSP1 in Q group were decreased compared with that of I groups(P<0.05).[Conclusions]QRYQTL preseription weakened the epithelial-mesenchymal transition (EMT) of podocyte by means of increasing the expression of P-cadherin and reducing the expression of FSP1, thereby decreased the Upro and protect the renal function.

OBJECTIVE:To investigate the significance of tacrolimus blood concentration monitoring to the therapy of mem-branous nephropathy. METHODS:41 patients with membranous nephropathy received tacrolimus,and the blood concentration of ta-crolimus reached to steady state. The trough concentration of tacrolimus was determined by EMIT. The patients were followed up, and clinical therapeutic efficacies were recorded. The relationship of blood concentration of tacrolimus with clinical efficacy was evaluated by SPSS 16.0 software. RESULTS:The blood concentration of tacrolimus was(7.47±2.74)ng/ml in complete remission (CR)group,(5.72±1.19)ng/ml in partial response(PR)group,and(3.30±1.08)ng/ml in no response(NR)group,with total remission rate of 75.61%. The blood concentration of CR group was the highest,followed by PR group and NR group,there was statistical significance among 3 groups(P<0.05). CONCLUSIONS:The clinical efficacy of tacrolimus in the treatment of nephrot-ic syndrome is correlate to the blood concentration intimately. Trough concentration monitoring of tacrolimus has important signifi-cance to the treatment of membranous nephropathy.

Objective:To investigate the relationship between gene polymorphisms of disease-relevant multiple cytokines including TNF-α,IL-6,IL-10,TGF-β1,IFN-γand acute graft versus host disease(aGVHD) in allogeneic hematopoietic stem cell trans-plantation ( allo-HSCT ) . Methods:32 cases of recipients received allo-HSCT and 36 cases of normal groups in January 2014 to December 2015 were selected as objects of study. We detected genotypes on specific SNP of target genes by polymerase chain reation ( PCR) combined with gene sequencing and observed the occurrence of aGVHD in postoperative recipients. The influence of cytokine gene polymorphisms on prognosis of allo-HSCT patients was analyzed,and the potential relationship between specific SNP mutation of the disease-relevant cytokine genes and severity of aGVHD was discussed. Results:Distribution of cytokines gene polymorphism including TNF-α-308(G/A),IL-6-174(G/C),IL-10-1082(A/G),TGF-β1+915(G/C),IFN-γ(T/A) had no significant differences with incidence of severe aGVHD(P>0. 05). However,the occurrence of severe aGVHD in allo-HSCT recipients with C/T genotype was significantly higher than C/C and T/T in SNP of TGF-β1+869(P<0. 01). Conclusion:Gene polymorphism of TGF-β1+869(C/T) in allo-HSCT patients was closely related to the occurrence of severe aGVHD. The research show allo-HSCT patients with C/T genotype occurred severe aGVHD more frequently, which is an important potential risk factor to induce the incidence of severe aGVHD. Therefore,detecting gene polymorphism of TGF-β1+869 ( C/T ) in allo-HSCT recipients and developing the appropriate therapeutic regimen may be helpful to reduce the incidence of aGVHD.

BACKGROUND:Chemotherapy drugs can damage the ovarian function in women of childbearing age, and even lead to premature ovarian failure. Therefore, to improve and restore the ovarian function in patients has become an important issue. OBJECTIVE:To explore the therapeutic effect and feasibility of bone marrow mesenchymal stem cel therapy against chemotherapy-induced ovarian damage. METHODS:Rat models of chemotherapy-induced premature ovarian failure were established, and injected with PKH26-labeled bone marrow mesenchymal stem cels. At 15, 30, 45, 60 days after cel transplantation, five rats were selected respectively to detect folicle-stimulating hormone and estradiol levels, and then, the rats were kiled to take the right ovary for pathological examination. The number of ovarian folicles was detected under light microscope. At 30 days after cel transplantation, another two rats were selected to mate with male rats to observe the difference in the reproductive activity. RESULTS AND CONCLUSION:Four of 22 rats (18%) gradualy recovered their estrous cycle after cel transplantation, with the decreased folicle-stimulating hormone level and increased estradiol level. Moreover, the number of folicles was reduced. Al of these indicated that the ability to have children in rats was not damaged.These experimental findings suggest that bone marrow mesenchymal stem cels can partialy improve the ovarian function of rats under chemotherapy.

AIM:To develop a capillary GC method for determining six components in Heiguidou Oil,?-pinene,eucalyptol,borneol,methyl salicylate,cinnamia aldehyde and eugenol. METHODS:GC was applied to quantitative analysis. Chromatographic conditions consisted of PEG-20M(30 m?0.25 mm?0.25 ?m),flame ionization detector(FID),and programmed temperature; initiation temperature was at 80 ℃ for 7 min; the rate of temperature was 15℃/minute,final temperature was at 180 ℃ for 10 min,bypass rate was 50 ∶1. The sample dissolved in absolute alcohol. The external standard was used. RESULTS:The standard curves of conclusion ?-pinene,eucalyptol,borneol,methyl salicylate,cinnamia aldehyde and eugenol were linear within the ranges of 0.322-32.150 mg/mL,0.202 -20.215 mg/mL,0.126-12.630 mg/mL,0.633-63.290 mg/mL,0.174-17.390 mg/mL,0.633-63.290 mg/mL and 0.203-20.255 mg/mL,respectively.RSD were 0.83%,0.48%,0.48%,0.31%,and 0.53 %,respectively. CONCLUSION:The established method is accurate and it can be applied to the simulta-neous assay of six components.

OBJECTIVE To explore occurrence, distribution,and resistance profile of ?-lactamases in clinical isolates of Enterobacter cloacae for rational use of antibiotics in clinic. METHODS Susceptibility test to 13 antibiotics was also performed through disk diffusion test.ESBLs were conformed according to NCCLS.DIDST test was adopted to detect AmpC ?-lactamases in E.cloacae isolates. RESULTS Among 78 isolates collected,13(14.9%) produced AmpC ?-lactamases,24(27.6%) produced ESBLs,10(11.5%) produced both ?-lactamases.These produced(?-lactamases) strains resisted to almost all ?-lactams,except for imipenem.The ?-lactamases producers possessed seriously multi-and cross-drug resistance. CONCLUSIONS ESBLs and AmpC ?-lactams are the main mechanism of E.cloacae resistance to antibiotic.

Objective To explore the effects of autologous blood transfusion on coagulation function,in-flammatory factors and immune function in patients with traumatic brain injury.Methods 85 patients taken autologous blood transfusion were selected as the observation group,30 cases with allogeneic blood transfusion were taken as control group.The blood transfusion volume,perioperative hematocrit,prothrombin time and postoperative complications were observed and compared between the two groups.Results In the observation group,the total amount of autologous blood transfusion in the observation group was 36 338mL,averaged (427.5 ± 28.3)mL,the total amount of allogeneic blood transfusion was 8 747 mL,averaged (102.9 ± 12.7) mL,and the autologous blood accounted for 80.6% of the total transfusion.61 patients only transfused autolo-gous blood,accounting for 71.8%.In the control group,the total allogeneic blood transfusion volume was 15 918 mL,averaged (530.6 ± 22.8)mL,the consumption of banked blood in the observation group was signifi-cantly less than that in the control group (P< 0.05).The levels of platelet (PLT) and plasma fibrinogen (FIB)in the control group increased significantly at 3 d after operation compared with those before operation (P<0.05);while activated partial thromboplastin time (APTT) began to decrease significantly at 3 d after operation (P<0.05);the prothrombin time (PT) and thrombin time (TT)decreased significantly at 7 d after operation(P<0.05),and the level of PLT and FIB in the observation group began to increase significantly at 1 d after operation (P<0.05).PT,APTT and TT were significantly reduced 1 d after operation.Compared with the control group,the level of PLT and FIB in each phase of the observation group was significantly high-er than that of the control group (P<0.05),and the PT,APTT and TT were significantly lower than those in the control group (P<0.05).After operation,the level of pro-inflammatory factors in the observation group was significantly lower than that of the control group.The level of anti inflammatory factors and immune function were significantly higher than that of the control group (P<0.05),and the incidence of complications in the observation group was 2.4%(2/85),which was significantly lower than 16.7%(5/30)of the control group,and the difference was statistically significant (P<0.05).Conclusion Autologous blood transfusion has the advantages of rapidness,timeliness and safety in the operation for patients with brain trauma.It is helpful to save blood,reduce the effect of trauma and blood transfusion on blood coagulation function and im-mune function,and reduce the risk of blood transfusion complications and postoperative infection,which is worth of clinical application.

Objective To investigate the effect of hyperthermia on human hepatocellular carcinoma cell line SMMC-7721 after down-regulating the expression of mTOR,and its possible mechanisms.Methods An antisense mTOR (mammalian target of rapamycin) gene eukaryotic expression vector was transfected into SMMC-7721 cells.The expression of mTOR mRNA and protein were detected using RT-PCR and Western blotting,respectively.Hyperthermia was applied after the transfection,and the vitality of cell proliferation was evaluated using CCK-8 assays and the clone formation rate was determined by colony-forming assays.The migration of SMMC-7721 cells was measured using scratch assays.Apoptosis and the cell cycle were analyzed by flow cytometry.Results The expression of mTOR mRNA and protein were significantly decreased after transfection,indicating that the antisense vector could down-regulate the mTOR gene effectively.The proliferation,clone formation and migration of SMMC-7721 cells all were decreased markedly by hyperthermia after transfection.Flow cytometry showed that the rate of apoptosis was significantly increased.The number of cells in the S phase was increased and the cell cycle was induced to arrest at the S phase.Conclusions Down-regulating the expression of mTOR can increase the thermosensitivity of SMMC-7721 cells.The mechanism involves increased apoptosis and S phase arrest.

Objective To provide reference for establishing diagnosis and differential diagnosis methods of rare yeast-like fungal bloodstream infection for clinical microbiology laboratory.Methods Trichosporon asahii (T.asa-hii)and Geotrichum capitatum (G.capitatum)bloodstream infection was diagnosed and differentially diagnosed through clinical data analysis,morphological examination,biochemical reactions,and molecular biology technology. Results Two types of yeast-like fungal bloodstream infections in case 1 and case 2 both occurred in leukemia agranulocytosis phase after chemotherapy,such infections were serious and highly similar.The cultivated colonies on blood agar plates of case 1 and case 2 were performed gram stain and microscopic examination.Hyphae,arthro-spores and microconidia were visible in the former,thickness of hyphae branches and length of arthrospores were different,most presented rectangular and barrel shape;the latter can be seen hyphae with transparent septum bro-ken up into arthrospores,presented rectangular shape,did not produce blastoconidia.Identification with API 20C AUX showed that they were T.asahii and G.capitatum.The PCR product sequences were compared with NCBI, suggesting that T.asahii and G.capitatum were at sexual stage.Conclusion Comprehensive application of a varie-ty of technical methods is helpful for improving the diagnosis accuracy of bloodstream infection with yeast-like fungi, identifying Trichosporon and Geotrichum to the species level may help physicians to understand such rare fungal in-fection,choose antifungal agents rationally,and improve clinical prognosis.