Objective To optimize the extraction technology for total flavonoids in Selaginella tamariscina.Methods To determine the content of total flavonoids by UV and the content of index constituent of amentoflavone by HPLC.The optimum extraction condition was investigated by orthogonal design and the extraction quantity was regarded as the investigated index.Results The optimum extracting condition was A1B2C2D2 with the extraction quantity of total flavonoids as the investigated index.The optimum extracting condition was A1～3B1～3C2D2 with the extraction quantity of amentoflavone as the investigated index.The optimum extracting condition was A1B2C2D2.That is adding ten times amount of 95% alcohol and refluxing twice,2 h once.Conclusion The optimum technology is stable and feasible for the extraction of S.tamariscina.

Objective To understand the molecular epidemiology characteristics and its drug resistance of methicillin-resistant Staphylococcus aureus (MRSA) in the urban area of Jilin and to provide important basis for guiding the clinical medication and prevention of the MRSA infection. Methods One hundred and three strains of MRSA from July 2013 to July 2014 in the urban area of Jilin were selected. The polymerase chain reaction (PCR) technology and multiple polymerase chain reaction were used to detect mecA gene and Staphylococcus chromosomal cassette mec typing (SCCmec) genotype of MRSA. The drug sensitivity test for 13 kinds of clinical common antibacterial drugs were detected by using the K-B method. And the source of the strains were analyzed. Results The results of SCCmec genotype of MRSA showed that SCCmecⅢtype were 62 strains, accounting for 60.2%;SCCmecⅡtype were 39 strains, accounting for 37.9%; failing to parting were 2 strains,accounting for 1.9%. Drug susceptibility test results showed that all of 103 MRSA strains were resistant to cefoxitin, cefazolin, penicillin and benzene, and drug resistance rate was 100.0%. The resistant rate to erythromycin, levofloxacin, ciprofloxacin, tetracycline, gentamicin and rifampin were 96.1%, 93.2%, 95.1%, 91.3%, 90.3%and 55.3%receptively;the resistant rate to sulfamethoxazolewas was only 1.9%;and the resistant strains to vancomycin and teicoplanin were not detected. The top three department of the distribution of the strains source were department of neurosurgery (31.1%), ICU (19.4%) and burn plastic surgery (17.5%). Conclusions The SCCmecⅢtype is the main MRSA epidemic strains, and SCCmec type II is a minor epidemic strainin the urban area of Jilin. The antibiotic resistance of MRSA is a serious problem with multiple drug resistance, but MRSA is sensitive to vancomycin and teicoplanin.

OBJECTIVE:To optimize the isolation and purification technology of active components of Senecio cannabifolius with macroporous adsorption resin. METHODS:The type of macroporous adsorption resin was selected with static adsorption using adsorption capacity and resolution rate of chlorogenic acid and hyperoside as index. The isolation and purification condition was op-timized with dynamic adsorption using the elution volume of chlorogenic acid and hyperoside as index,such as maximal sample size,rinse water quantity,volume fraction and collective volume of eluant ethanol. RESULTS:Among 7 kinds of resin,HPD100 had the best purification property;the optimal purification technology was as follows as mass concentration of sample 6 mg/ml,the speed of sample loading 2 ml/min,maximal sample size 3 times of column volume(BV), rinse water quantity of 2.5 BV,60%ethanol as eluting reagent. The contents of chlorogenic acid and hyperoside were increased from 0.90,0.18 mg/g to 7.26,1.04 mg/g after purification. RSD of each index were all ≤3.0%(n=3) in validation test. CONCLUSIONS:The isolation and purification technology of active components of S. cannabifolius with HPD100 macroporous adsorption resin is stable and effective.

Objective To establish a method to induce dendritic cells (DC) from peripheral blood mononuclear cells of healthy people in normal human AB serum in vitro and to identify the phenotype and the function of DC. Methods Peripheral blood mononuclear cells (PBMNC) of healthy people were cultured in RPMI 1640 media including human AB serum, GM-CSF, rhIL-4, and/no rhCD40 for 7 days to generate DC,which were identified by morphological features, surface antigen expression and the ability to stimulate T cells.Results After cultured and induced, DC displayed typical morphology with elongated dendritic process viewed by inverse light microscope as well as Wright-Gimsa stain. Mature DC express CD83 and the costimulatory molecules CD40 CD80, and CD86 to effectively activate T cells. In the five time points of 0 day, 1st day, 3rd day, 5th day and 7th day, the expression of CD83, CD40, CD80, CD86 and CD14 were significantly different (F= 50.253, 243.769, 248.181, 191.267 and 226.339, respectively, P＜ 0.05). The ability to stimulate T cells in GM-CSF, rhIL-4, and rhCD40L group was also stronger than that in GM-CSF and rhIL-4 group. DC started to secrete IL-12 from 5th day, the values were (42.92±1.54) pg/ml and (136.18±5.27) pg/ml in group of plus CD40L and of non plus CD40L, respectively. The secretion of the two groups of IL-12 were (60.09±2.27) pg/ml and (322.30±30.60) pg/ml (t = -44.941, -22.611, bath P ＜ 0.05). There are significant differences between the two groups. Conclusion DCs can be cultured from the peripheral blood of healthy people in normal human AB and rhCD40L serum.

OBJECTIVE:To develop a method for simultaneous determination of chlorogenic acid,geniposide,gentiopicro-side,ferulic acid,baicalin and ammonium glycyrrhizinate in Yindan pinggan capsule. METHODS:HPLC method was adopted. The separation was performed on Wonda SilTM-C18 column with mobile phase consisting of acetonitrile-0.4% phosphoric acid solu-tion(gradient elution)at the flow rate of 0.8 mL/min. The detection wavelength were set at 325 nm(chlorogenic acid,ferulic ac-id),250 nm (geniposide,ammonium glycyrrhizinate) and 275 nm (gentiopicroside,baicalin). The column temperature was 30 ℃. RESULTS:The linear ranges were 0.087-3.480 μg for chlorogenic acid(r=0.9998),0.201-8.040 μg for geniposide(r=0.9997),0.200-8.000 μg for gentiopicroside(r=0.9995),0.016-0.640 μg for ferulic acid(r=0.9999),0.105-4.200 μg for ba-icalin (r=0.9999) and 0.028-1.120 μg for ammonium glycyrrhizinate (r=0.9995),respectively. The limits of quantitation were 1.31,0.75,1.14,1.25,0.94,0.98 ng,and the limits of detection were 0.87,0.67,0.96,0.93,0.60,0.88 ng,respectively. RSDs of precision,stability and reproducibility tests were lower than 2.0%;recoveries were 99.9%-101.9%(RSD=0.7%,n=6), 98.7%-100.9%(RSD=0.9%,n=6),98.1%-101.5%(RSD=1.4%,n=6),98.5%-101.3%(RSD=1.3%,n=6),98.5%-101.7%(RSD=1.2%,n=6),98.2%-101.4%(RSD=1.2%,n=6),respectively. CONCLUSIONS:The method is simple and accurate , can be used for simultaneous determination of chlorogenic acid,geniposide,gentiopicroside,ferulic acid,baicalin and ammonium glycyrrhizinate in Yindan pinggan capsule.

Objective The purpose of this study was to discuss the therapies for hemorrage caused by the fissuration of pancreatojejunal stoma and pancreatic leakage after pancreatoduodenectomy.Methods After three cases of pancreatoduodenectomy,the disruptions of pancreatojejunal stoma resulted in serious pancreatic leakage and the hemorrage in abdominal cavity.During all the second operations,the drainage-tube insertions into the main pancreatic ducts were used to lead the pancreatic juice into the neighboring loop of jejunum.Results Afer the operations,the supportive treatment,continuous irrigation of peritoneal cavity and pancreatic enzyme inhabition were given to the patients of these cases and all of the patients were successfully cured.Conclusions The bridge-crossing internal drainage which inserts drainage-tube into the main pancreatic duct was a convenient and effective therapy and method to rescue the hemorrage caused by the fissuration of pancreatojejunal stoma and pancreatic leakage after pancreatoduodenectomy.While the patients' lives were saved,their functions of pancreas were preserved and the qualities of life were improved after the operations.

Objective To explore the expression level of thioredoxin (Trx) in acute myeloid leukemia (AML) cells,and its association with clinical features and prognosis,as well as the effects of diamide,an inhibitor of Trx,in inducing AML cells apoptosis.Methods Expression of Trx on AML cells and fresh bone marrow cells from healthy adults were analyzed by Western-blotting. The inhibition of proliferation was measured by MTT assay.The anti-leukemia effect of diamide was observed by morphology and agarose gel electrophoresis.Results 75 ％ (15/20) of AML patients expressed Trx,while no expression was observed in control group.Higher expression level of Trx was associated with higher WBC counts (x2 =9.375,P ＜ 0.05),which suggested that overexpression was associated with leukemogenesis.The inhibition of diamide on AML cells showed time and dose dependent by MTT assay.The IC50 values of diamide at 24 h,48 h and 72 h were 98.26 mg/ml,47.53 mg/ml and 8.34 mg/ml,respectively.After AML cells were treated with diamide,the apoptotic body appeared by morphology,and the typical DNA “ladder” bands were confirmed by agarose gel electrophoresis.Conclusion Trx expression level in AML cells is significantly higher than that of control group.Diamide inhibites the proliferation of AML cells by inducing apoptosis,which might be a potential agent for AML.

Objective To investigate the infection status quo and genotype distribution of human papillomavirus(HPV) infection situation among females in west Guangxi area.Methods Cervical exfoliative cells samples from 3 315 women were collected to detect HPV genotyping with Cape flow-through hybridization.Then the results were statistically analyzed.Results The overall HPV infection rate was 21.30%(706/3 315),in the females of HPV positive infection,the high-risk type infection was predominant,accounting for 89.52%(632/706).The HPV infection type was dominated by single type infection,accounting for 72.66% the double infection accounted for 22.10%(156/706). The 21 HPV subtypes were detected.The high-risk HPV subtypes with high detection rate were HPV52(26.77%),HPV16(15.30%) and HPV58(15.01%).The low-risk HPV subtypes with high detection rate were HPV CP8304(11.90%) and HPV6(3.68%).The HPV subtypes were distributed differently at different ages.In 7 age groups of≤20,>20-30,>30-40,>40-50,>50-60,>60-70,>70 years old,the infection rates of high risk HPV were 21.62%(8/37), 19.26%(120/623),17.66%(220/1 246), 14.88%(153/1 028), 16.83%(51/303),15.52%(9/58) and 30.00%(6/20) respectively,showing no statistically significant differences among them(χ2=10.019,P=0.124).Conclusion Cervical HPV infection are mainly high-risk HPV subtypes and single type infection in females of western Guangxi area.The HPV subtypes with high infection rate are 52,16,58 and CP8304.

Objective The purpose of this study was to determine how to preserve the remaining pancreatic body and tail in the pancreatectomy. Methods In seven cases of pancreatectomy, three of them were the rupture of pancreatojejunal anastomosis, and four of them were the pancreatectomy for tumor in the pancreatic neck or body. During operations, a bridge internal drainages was used to drain the pancreatic juice into the adjacent jejunum. After the operations, the supportive treatment, continuous irrigation of peritoneal cavity and pancreatic enzyme inhibition were used. Results In all seven patients, the remaining pancreatic body and tail were successfully preserved. The endocrine functions of these patients recovered to nearly normal level and patients were discharged. Conclusions In preserving the remaining pancreatic body or tail, the bridge internal drainage has its advantage of convenience. It effectively preserves the exocrine of pancreas as well as its endocrine

ObjectiveTo induce monocyte-derived dendritic cells(MoDC)from acute myeloid leukemia (AML) and healthy persons by rhCD40L in normal human AB serum system in vitro and to identify the morphology and phenotype of MoDC. MethodsPeripheral blood mononuclear cells(PBMNC)of AML and healthy persons were cultured in RPMI 1640 media including human AB serum, GM-CSF, rhIL-4 and rhCD40L, respectively. MoDC were identified by morphological features, surface antigen expression and the ability to stimulate T cells. ResultsAfter cultured for 7 days, MoDC displayed typical morphology with elongated dendritic process,and upregulation of the costimulatory molecules CD40,CD80,CD86 and CD83.The morphology and expression of costimulatory molecules were not significantly different between AML and healthy persons (P＞0.05),but were significantly different between rhCD40L group and without rhCD40L group (P＜0.05). MoDC had the ability to activate T cells, and there were no statistical differences between AML and healthy persons(P ＞0.05), but were significant differences between rhCD40L group and without rhCD40L group (P＜0.05). MoDC started to secrete IL-12 on day 5, and there was no statistical differences between AML and healthy persons(P＞0.05),and had differences between rhCD40L group and without rhCD40L group (P＜0.05).ConclusionMoDC can be cultured from the peripheral blood of AML and healthy persons.There were no significant differences in morphology and phenotype.Monocyted-derived DC can be used as an alternative to generate leukemia-specific cytotoxic T cells,especially in the presence of rhCD40L.