Objective:To establish an HPLC method for the determination of paclitaxel and docetaxel in plasma to provide refer-ence for the individualized treatment regimen and the evaluation of curative effect and adverse reactions. Methods:Paclitaxel and do-cetaxel were used as the internal standard for each other. The samples were precipitated by acetonitrile and separated on a DikMA Dia-monsil C18 column with a mixture of acetonitrile-water (55: 45) as the mobile phase. The flow rate was 1. 2 ml·min-1 . The column temperature was set at 25℃. Paclitaxel and docetaxel were detected by UV-detection (λ= 227 nm). Results: A linearity was ob-tained within the range of 0. 078-10. 0 mg·L-1 for paclitaxel and docetaxel. The limit of quantitation was 0. 039 mg·L-1 . The aver-age recovery of paclitaxel and docetaxel was 99. 85% and 100. 35%, respectively. The inter- and intra-day RSD were both less than 5% and the RSD for freeze-thaw stability was below 10%. The plasma concentration of paclitaxel in clinical samples was within the range of 0. 18-6. 16 mg·L-1 and obvious individual difference was shown. Conclusion:Therapeutic drug monitoring is very important due to the obvious differences in plasma concentration of paclitaxel and docetaxel. The established method is sensitive, accurate, con-venient and rapid in r the therapeutic drug monitoring, and is useful for the adverse drug reactions monitoring and pharmacokinetic study.

Objective To evaluate the therapeutic effect of lentinan combined with conventional antituberculous therapy in women with pelvic tuberculosis.Methods A total of 90 women with pelvic tuberculosis were enrolled and recruited into a control group and a treatment group by random number table,45 in each group.The control group was treated with anti-tuberculosis program (2HRZE/4HRE),while the treatment group was treated with lentinan tablets on the basis of the control group.Both groups were treated for 6 months.Serum levels of carbohydrate antigen 125 (CA125) and erythrocyte sedimentation rates (ESRs) were determined before and after treatment,and the therapeutic effect was evaluated.Results After treatment,the serum CA125 level (28.61 ± 9.08 U/ml vs.39.72 ± 12.13 U/ml;t=4.919,P＜0.01) and the ESR (36.13 ± 8.33 mm/h vs.41.35 ± 12.45 mm/h;t=2.338,P＜0.05) in the treatment group were significantly lower than those in the control group.The negative rate of serum CA125 after treatment than before treatment in the treatment group (93.3％ vs.20.0％;X2=46.335,P＜0.01) and the control group (82.2％ vs.8.9％;X2=37.396,P＜0.01);but there was no difference in negative rate of serum CA125 after treatment between two groups (X2=1.6571,P=0.198).The total effective rate in the treatment group was significantly higher than that in the control group (93.3％ vs.77.8％;X2=4.406,P=0.036).Conclusion Lentinan combined with conventional antituberculous therapy is effective in treatment of pelvic tuberculosis in women.

In order to strengthen the management of disinfection quality of endoscopes, Quality Control Center of Digestive Endoscopy and Nosocomial Infection Control Center of Yunnan Province investigated the diagnosis, treatment, cleaning and disinfection conditions and disinfection quality of digestive endoscopes in some medical institutions of Yunnan Province by web questionnaire from April to May in 2019, and 277 valid questionnaires were finally obtained. SPSS 19.0 statistical software was used to analyze the influencing factors of cleaning and disinfection process and the infection control implementation of digestive endoscopes in 227 secondary and tertiary hospitals. The results showed that the number of decontamination people who had received systematic training in Yunnan Province was significantly lower than that in other areas of China. The hospital level, the number of decontamination personnel, and decontamination methods affected the implementation of cleaning and decontamination process and infection control, while the allocation of decontamination supplies had no effects. It is important to establish an effective mechanism for the normalized implementation of cleaning and disinfection of digestive endoscopes.

Objective:To analyze the effect of continuing care in patients treated with endonasal endoscopic conjunctivorhinostomy, so as to guide its clinical application.Methods:A total of 192 patients were selected with endonasal endoscopic conjunctivorhinostomy who were admitted to the Aier Eye Hospital of Wuhan University from June 2013 to June 2019 as the research objects. Randomized digital table method was used to divide them into control group and observation group, 96 cases in each group. The control group was given routine instructions before discharge, while the patients in observation group were given continuing care intervention on the basis of control group, including establishment of a continuing care group, establishment of health files of discharged patients, regular follow-up through telephone, establishment of WeChat groups for patients or their families, opening WeChat official account, organization of doctor-patient discussion meeting, and setting up reexamination services. The reexamination adherence rate, medication adherence rate, care satisfaction and treatment efficacy were compared between the two groups 6 months after implementation.Results:The reexamination adherence rate of the observation group on 1, 3, and 6 months after discharge were 85.42%(82/96), 96.88%(93/96), 72.92%(70/96), and the control group were 62.50%(60/96), 73.96%(71/96), and 43.75%(42/96), respectively, the difference between the two groups was statistically significant ( χ2 value was 13.088,20.237, 16.800, P<0.05); the medication adherence rate of the observation group was 89.58%(86/96), and the control group was 62.50%(60/96), the difference between the two groups was statistically significant ( χ2 value was 19.326, P<0.05); the observation group's care satisfaction was 94.79%(91/96), the control group was 78.13%(75/96), the difference between the two groups was statistically significant ( χ2 value was 11.388, P<0.05); the total treatment efficacy in the observation group was 96.88%(93/96), and the control group was 87.50%(84/96), the difference between the two groups was statistically significant ( χ2 value was 5.858, P<0.05). Conclusion:The application of continuing care for patients treated with endonasal endoscopic conjunctivorhinostomy can improve patients′ reexamination and medication adherence, improve care satisfaction, promote patients′ good recovery and achieve better surgical effects, which is worthy to recommend its clinical application.

Objective:To construct the eukaryotic expression plasmid carrying hTERT-P2A-EGFP, and to explore its expression and transfection efficiency in the HEK293FT cells.Methods:The recombinant plasmid was constructed by using pBABE-puro-hTERT and pRRLSIN-cPPT-MSCV-EGFP plasmids.The hTERT,P2A,and EGFP genes were obtained using pBABE-puro-hTERT as template by PCR.And the correct hTERT was inserted into pRRLSIN-cPPT-MSCV-EGFP vector.Then the recombinant plasmid containing hTERT-P2A-EGFP gene was obtained and identified.The HEK293FT cells were transfected by the recombinant plasmid, and the expression of green fluorescence protein(GFP) was observed by fluorescence microscope.Results:The PCR results showed that the fragments of hTERT, P2A, and EGFP were 3 400, 110 and 720 bp.And the length of gene fragment(hTERT-P2A-EGFP)was 4 300 bp by enzyme digestion.The results of sequencing showed that the 1 547 site of the target gene was mutated.Using site-directed mutagenesis, the 1 547 site was successfully mutated.And the target gene sequence was completely identical with the sequence published in GenBank.The recombinant plasmid was transfected into the HEK293FT cells, and GFP was observed in the cells.The results of flow cytometry showed that the transfection efficiency of recombinant plasmid was 44.8%.Conclusion:The recombinant plasmid carrying hTERT-P2A-EGFP gene is successfully constructed, and it can be used for cell transfection.

Objective To explore the potential mechanisms of non-small cell lung carcinoma cells to rsTRAIL protein-induced apoptosis by phosphatidylinositol 3′-kinase (PI3K/Akt)inhibitor LY294002,and to provide new ways to increase killing activities of rsTRAIL protein for non-small cell lung cancer.Methods The A549 cells at logarithmic growth phase were selected and randomly divided into rsTRAIL group and LY294002+rsTRAIL group. The inhibitory rate of growth of the A549 cells was tested by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry analysis. The expression levels of Ser473 phosphorylated form of Akt (p-Akt),c-FLIPL protein and Bcl-2 protein in the A549 cells in two groups were analyzed by Western blotting method. Results The inhibitory rate of growth of the A549 cells in LY294002+rsTRAIL group (74.6 %± 2.63%)was higher than that in rsTRAIL group (5.61% ± 0.32%) (P< 0.05 ). Compared with rsTRAIL group, the percentage of the cells at G0/G1 phase in LY294002+rsTRAIL group was increased(P<0.05)and the percentage of the cells at S phase was decreased(P<0.05).The apoptotic rate of the A549 cells in LY294002+rsTRAIL group (61.5%±3.02%)was higher than that in rsTRAIL group (3.21%±0.96%)(P<0.05). The Western blotting results showed that the expression levels of p-Akt, c-FLIPL and Bcl-2 proteins in the A549 cells in LY294002+rsTRAIL group were decreased (P<0.05 )and the ratio of Bax/Bcl-2 was increased (P<0.05 ) compared with rsTRAIL group.Conclusion LY294002 can increase the killing activity of rsTRAIL protein in A549 cells by inhibiting the activity of PI3K.

Objective To explore whether synergistic action occurs in development of depression-like behaviors using ovariectomy combined bondage stress,and whether hippocampal neural progenitor state is changed following depression-like behaviors in mice.Methods Forty female C57B/L mice were divided into four groups randomly,which included ovariectomy,bondage stress,ovariectomy combined bondage stress and sham groups.Depression-like behaviors were evaluated by open field test,sucrose preference and forced swimming test after animals had been treated for 21 days.All animals were fixed by 4％ PA perfusion through left ventricle,then brain was removed,and coronal serials sections were made.Expression of Ki67 and DCX was tested by immunohistochemistry or immunofluorescence in subgranular zone of hippocampal dentate gyrus (SGZ).Results Body weight((18.70± 0.25) g vs (20.96±0.24)g),novelty-seeking behavior and sucrose preference decreased in ovariectomy combined bondage stress-group compared with those in sham group (P＜0.05),but forced swimming time increased in shame group((165.6±9.6)s vs (140.28±12.3) s).Likewisely,Ki67-positive cells((8.6±2.4)/section) and DCX-positive cells((4.2±1.4)/section) in SGZ decreased in ovariectomy combined bondage stress-group compared with those in sham group(Ki67:(16.7±2.5/section),DCX:(12.6±2.3/section) (P＜0.05).Unsimilarly,depressionlike behaviors had little change in ovariectomy-or bondage stress-groups compared with those in sham-group.Conclusion Ovariectomy combined bondage stress has synergistic action in development of depression,and inhibits the active state of progenitor in SGZ in mice.

Objective:To observe the effect of tannic acid on renal morphology and function of diabetes mellitus model rats ,and to explore the mechanism of improving effect of tannic acid from oxidative stress , nitrosative stress angle.Methods: 8 rats were randomly selected from 68 6-week-old male Wistar rats as normal control group and the remaining 60 rats accepted high-sugar and high-fat diet for 4 weeks, then were injected streptozotocin ( STZ, 52 mg/kg ) intraperitoneally in order to manufacture a diabetic rat model.Further the diabetic rats were randomly divided into model group ,aminoguanidine group ,low-dose of tannic acid group and high dose of tannic acid group.The rats in aminoguanidine group were injected aminoguanidine [AG,40 mg/(kg· d)] intraperitoneally, those in low-dose of tannic acid group were injected tannic acid [TA,20 mg/(kg· d)] and those in high-dose of tannic acid group were injected tannic acid [TA,30 mg/(kg· d)].The rats in normal control group and model group were injected normal saline [NS, 30 mg/(kg· d)] and all rats were sacrificed and tissues were derived at the end of the week 10.Morphologic changes of kidney in diabetic rats were observed by HE staining and correlative biochemical indices of renal function were detected by biochemical analyzer.8-hydroxy deoxyguanosine (8-OHdG) and 3-nitrotyrosine (3-NT) content of renal tissue in rats in different groups were detected by ELISA method.Mesangial cells cultured in vitro were treated with high concentration of glucose (30 mmol/L) and AGEs (250 mg/L) and at the same time with different concentration of tannic acid (10,20,40 and 80μmol/L) on the basis of setting corre-sponding control group.The contents of 8-OHdG and 3-NT in the culture supernatant were measured by ELISA method after 48 hours.Results:Tannic acid can effectively improve the renal pathological changes and improve renal function of diabetic rats .The contents of 8-OHdG and 3-NT in kidney tissue homogenate of diabetic rats and in the supernatant of GMC cultured with high glucose or AGEs were all significantly increased and can be reduced by tannic acid.Conclusion:Tannic acid improving the structure and function damage of kidney in diabetic rats might be achieved by oxidative stress and nitrosative stress mechanism .

Objective To clarify clinical and morphological features and immunophenotype and Epstain-Barr virus infection of neoplastic cell rich Hodgkin's lymphoma (NCRHL)and to further improve our knowledge and pathological diagnosis for NCRHL. Methods 10 cases of NCRHL were analyzed for clinical features, morphology, immunophenotype, Epstein-Barr virus infection using routine eosin and haematoxylin stain, immunohistochemistry, Epstain-Barr virus encoded small RNA (EBER) in situ hybridization and combining clinical data. Results (1)NCRHL were more common in young people. The median age of the patients was 25.5 years old. The ratio of male to female was 1:2.3. Superficial lymph nodes were most frequently involved. Masses of mediastinum were seen commonly. Clinical manifestation of the patients included B symptom (6 cases), pruitus (5 cases) and anemia (1 case). (2)Architecture of lymph nodes were effected. Necrosis was seen in some cases. There were more tumor cells in NCRHL than that in the classical Hodgkin's lymphoma. The tumor cells were distributed in piece or patch or diffuse. The morphology of neoplastic cells was wore variable including Hodgkin-like cells, lacunar cell-like, mummy cell-like and anaplastic large cell-like, singular nucleated cells, and multinucleated giant cell-like cells. Numerous neutrophils and eosinophils were present in a few cases. Focal sheet, necrosis granulomatosis-like and diffuse growth pattern were found in NCRHL. (3)All of the cases were positive for CD30 and PAX-5.2/10 (20%) cases were CD15 positive. LCA, CD20 and CD3 were negative. (4)EBER was not detected in all 6 tested cases. (5)Follow up data was obtained in 8/10 cases, in which one patient was dead, one case relapsed in half a year,and the other 6 cases reached complete regression. Conclusion NRCHL is characterized mainly by neoplastic cell rich morphologically and focal sheet, necrosis granulomatosis-like and diffuse growth pattern.EBER was not detected in this tumor. Some cases have aggressive clinic process with a unfavourable prognosis. New treatment regimen should be explored.

Objective To investigate the anti-oxidant effect and the possible mechanisms ofpicrodide II in cerebral ischemia/reperfusion injuries in rats. Methods A total of 90 adult, healthy, mmale Wistar rats were used to established the middle cerebral artery occlusion reperfusion (MCAO/R) models by intraluminal monofilament suture on the left external-internal carotid artery. The treatment group and the positive control group were respectively injected with 1.0％ picroside II (10 mg/kg, 250 μl) and salvianic acid A sodium (10 mg/kg, 250 μl) via the tail vein, and the negative control group and sham-surgery group were injected with 0.1mol/L phosphate buffer saline (PBS) 250 μl. The neurological deficit scores were evaluated with Bederson's test. The cerebral infarction volume was observed with tetrazolium (TTC) staining. The apoptosis positive cells were counted by terminal deoxynucleotidyl transferase dUTP nick-end labeling and the expressions of inducible nitric oxide synthase (iNOS) and superoxide dismutase (SOD) were detected with immunohistochemical assay.The concentration of iNOS and SOD proteins in brain tissue was detected by enzyme linked immunosorbent assay.Results Neurological behavioral malfunction appeared in all the rats with MCAO/R. The infarction focuses emerged in the ischemic hemisphere following the MCAO/R injuries. The number of apoptotic cells and the expression of iNOS increased while the SOD reduced after MCAO/R. After the treatment of picrodide Ⅱ, the nervous behavioral function (1.28±0.38)improved, the infarction volume(68.73±4.46)％ reduced, the number of apoptosis positive cells(6.10± 1.26), the expressions and the concentrations in brain tissue of iNOS(4.67+0.51)decressed while those of SOD (0.53 ±0.14) increased significantly compared with the negative control groups(t=3.16、 2.51、 4.15、3.12、 3.25, P＜0.05). Conclusion PicrodideⅡ might play a neuroprotective effect by inhibiting the neuronal apoptosis and the expressions of iNOS and SOD after cerebral ischemia/reperfusion injuries.