OBJECTIVE:To establish a method for the determination of 100 pesticide residues in Morus alba. METHODS:LC-MS/MS was conducted on the column of ZORBAX Eclipse plus C18 with mobile phase of acetonitrile (containing 5% water, 0.1% formic acid,5 mmol/L ammonium formate)- water(containing 0.1% formic acid,5 mmol/L ammonium formate)(gradient elution)at a flow rate of 0.4 ml/min,column temperature was 40 ℃,injection volume was 5 μl;MS conditions:ionization source was electrospray ionization with positive ion mode,scanning mode was dynamic MRM,scanning time window was 2 min;atomiz-ing gas was nitrogen,atomizing gas pressure was 40 psi,drying gas flow rate was 5 L/min,capillary voltage was 4 000 V,and ion spray voltage was 500 V. RESULTS:The linear range of 100 pesticide residues was 2.4-150 ng/ml(r>0.990 0),recovery was 69.3%-128.2%,and the determination limit was 0.003-16 μg/kg. CONCLUSIONS:The method is simple,stable and reproduc-ible,and can be used for the determination of pesticide residues in M. alba.

Objective In order to study the biological significance of HBV/C gene variant in vitro.Methods Retroviral vector pXT1 was used to construct the HBV/C gene expression vector and the recombinant plasmid pXT1-HBV/C was used to transfect immortalized human peripheral blood B cell lines to express HBcAg steadily in the host cells.Results plasmid pXT1-HBV/C was detected positive by PCR as well as enzyme digestion with Bgl Ⅱ and Xho Ⅰ.Meanwhile.transfected cells was detected to contain HBV/C gene by PCR and HBcAg was expressed in 47.4%of the ceils by means of flow cytometry.Conclusion Retroviral expression vector with HBV/C gene can transfeet into eucaryotic cells effectively and express the goal gene steadily.The recombinant cells may be used in the systematic studies on the biological significance of HBV/C gene variant.

Objective To explore the efficacy and safety of human recombinant vascular endothelial inhibition (en) combined with gamma knife in the treatment of elderly advanced non-small cell lung cancer patents.Methods Totally 38 cases of senile non-small cell lung cancer patients were divided into two groups:A group was treatment group with 17 cases of recombinant human endostatin combined with gamma knife,group B was control group with 21 cases of patients receiving body gamma knife alone.Recombinant human endostatin was given intravenously 15 mg per day in 500 ml of saline for 3 ～ 4 hours for 14 days a cycle,and the total dose of body gamma knife therapy was 36 ～ 48 Gy/10 ～ 12f.Results The response rate (RR) of groups A and B were 58.82％ and 57.14％ without statistical difference.One-year survival rate of groups A and B were 47.06％ and 38.09％,respectively;and there was no statistical significance.The main side-effects of both groups were bone marrow suppression and gastrointestinal reactions,but no statistical difference was observed.Two groups of patients'quality of life were improved without statistical difference (P ＞ 0.05).Conclusions Body-gamma knife therapy combined with recombinant human endostatin in elderly advanced non-small cell lung cancer patients has better efficacy such as CR rate and long-term survival,and similar side-effects.

Objective To study the effects of extensively hydrolyzed formula forvery/extremely low birth weight(VLBW/ELBW)infants.Methods From Jun.2013 to Oct.2015,78VLBW/ELBW infants admitted to our hospital within 12 hours of birth were randomly assigned into two groups:hydrolyzed protein formula feeding group ( hydrolyzed formula group) and preterm formula feeding group (control group). Infants with hospital stay <28 days were excluded. Prospective study was conducted between two groups comparing the duration of meconium discharge, the time required for total enteral feeding average hospital stay, feeding intolerance and physical growth and blood biochemical indices on the28thday.Results A total of 78 infants were enrolled,35 in hydrolyzed formula group and 43 in control group. Comparing with control group, feeding intolerance in hydrolyzed formula group was significantly lower ( 25. 7℅ vs. 72. 0℅) . The duration of meconium discharge [ ( 4. 9 ± 0. 8 ) d vs. (8. 8 ± 1. 6)d], the time required for total enteral feeding[(13. 4 ± 2. 0) d vs. (18. 9 ± 2. 6) d] and average hospital stay duration [ ( 33. 7 ± 5. 1 ) d vs. ( 41. 8 ± 6. 8 ) d ] was shorter in hydrolyzed formula group ( P<0. 05). The body length on the 28th day in hydrolyzed formula group was longer than control group[ (43. 8 ± 1. 2 ) cm vs. ( 42. 6 ± 2. 0 ) cm, P < 0. 05 ] . The concentration of serum albumin [ (32. 5 ± 3. 0 ) g/L vs. ( 30. 0 ± 4. 5 ) g/L ] and hemoglobin [ ( 112. 4 ± 11. 4 ) g/L vs. ( 106. 3 ± 13. 0) g/L] in hydrolyzed formula group were significantly higher than the control group( P<0. 05). No significant difference was found between the two groups regarding the time required returning to birth weight [(10. 9 ±2. 2)d vs. (10. 1 ±1. 7)d],body weight [(1759 ±107)g vs. (1627 ±435)g], head circumference[(30. 3 ± 1. 0)cm vs. (29. 7 ± 1. 6)cm] on the 28th day and the incidence of extrauterine growthretardation(EUGR)(40.0℅vs.44.1℅)atdischarge.Conclusions Comparing with preterm formula feeding, early feeding with extensively hydrolyzed formula can reduce the incidence of feeding intolerance on VLBW/ELBW infants. Extensively hydrolyzed formula also can accelerate meconium discharge, reduce hospital stay duration and the time required for total enteral feeding. But the growth of weight and head circumference on the 28th day and the incidence of EUGR at discharge were not altered by extensively hydrolyzed formula.

AIM: To investigate the potential role of angiopoietin-2 in the rats model of lipopolysaccharide (LPS)-induced acute lung injury (ALI). METHODS: Wistar rats were randomly divided into four groups(n=10): control group; in LPS groups, rats were divided into three subgroups, treated with different dosage of LPS (2 mg/kg, 4 mg/kg and 8 mg/kg). ALI model was established by intravenous injection with LPS, while control group was with NS. The lung histopathology change was observed by HE stain. Western blot was used to measure the expression of Ang-2 in serum. RESULTS: Histologically, alveolar edema, hemorhag, and massive inflammatory cell, infiltration were observed in LPS groups, but not in control group. The pathological score in LPS groups were significantly higher than that in NS group. Compared with 2 mg/kg LPS group, the pathological score in 4 mg/kg LPS group was significantly higher. While the pathological score in 8 mg/kg LPS group was higher than those in other LPS groups. The expression of Ang-2 protein in plasma was significantly higher in LPS groups, but weak in control group. Compared with 2 mg/kg LPS group, the expression of Ang-2 protein in 4 mg/kg LPS group was significantly higher. While the expression of Ang-2 protein in 8 mg/kg LPS group was higher than those in other LPS groups.The expression of Ang-2 protein in plasma were significantly positively correlated with the pathological score (r=0.862, P<0.05). CONCLUSION: Ang-2 could participate the pathological course in the rats model of LPS-induced acute lung injury. The expression of Ang-2 protein in serum was significantly positively correlated with the extent lung injury.

OBJECTIVE: To establishes an HPLC method for determination of the contents of Osthol and Berberine in Kangfuyan effervescent suppository. METHODS: HPLC was carried out on a column of Diamonsil (250 mm?4.6 mm,5 ?m) with mobile phase consisted of acetonitrile-0.2% phosphate acid (52∶48,Lauryl sulfonic acid sodium 0.1 g was contained in every 100 mL) with flow rate at 1 mL?min-1.The detection wavelength was set at 322 nm and the column temperature was set at room temperature. RESULTS: The linear range was 0.019 8～0.396 0 ?g(r=0.999 97) for Osthole versus 0.056 65～1.133 00 ?g(r=0.999 99) for Berberine. The average recoveries of Osthol and Berberine were 101.06% (RSD=1.10%) and 101.88%(RSD=0.98%), respectively. CONCLUSION: This method is simple, accurate and reproducible, and applicable for the quality control of Kangfuyan effervescent suppository.

Objective To study the biological significance of the common mutant preC/C gene in clinical HBV in China. Methods Site directed mutagenesis based on the unique enzyme site elimination was used to construct eukaryocyte expression vector with mutant HBV/C gene(V60、G87、L97). Expression vectors with wild and mutant preC/C gene were transferred into HepG2 cell. Culture supernatant was detected by ELISA for HBeAg. Results Result of DNA sequencing showed that the constructed mutant HBV preC/C gene had only one specific site variation compared with the wild type sequence. Goal DNA fragment was detected positive in the HepG2 cells transferred with wild and mutant preC/C gene. A value of HBeAg in the supernatant of the cells harboring L97 variant was higher than that of the wild and other variant strains( P

To study the characteristics of EBV transformed human peripheral blood B cell lines from hepatitis B patients and to provide a basis for further studies on the cellular immune function of hepatitis B patients.High titer of EBV was produced by B95 8 cell by induction with sodium n butyrate,and peripheral mononuclear cells from two different groups of hepatitis B patients were harvested and infected with EBV. The results showed that 4～8 weeks after infection 19 EBV transformed B cell lines from hepatitis B patients were established. It was found that colony formation of cells from A group appeared 2～3 weeks later than B group. Cells grew healthily and had good activity after 4 weeks of freezing.CD19 and CD20 were detected in 88 34% and 37 48% of the immortalized B cell membrane respectively. HBV DNA could not be detected in a11 19 immortalized B cell lines.It suggested that the time needed for the establishment of B cell lines was related with the immune state of the patients.HBV DNA could not exist persistently, and it would disappear finally in the immortalized B cell lines.

Objective To evaluate the clinical efficacy of laparoscopic surgery in the treatment of endometriosis. Methods Clinical data of 102 patients with endometriosis treated by laparoscopic surgery were retrospectively analyzed. Results There were no conversions to open surgery or complications in all the patients. The operation time was 80 5 min ? 28 3 min, the intraoperative blood loss was 25 2 ml ? 23 2 ml and the postoperative hospital stay 3 0 d ? 1 2 d. Follow-up for 6 months ～ 66 months in 83 patients showed 21 cases of recurrence. Normal pregnancy was seen in 23 patients among 53 patients with endometriosis associated with infertility. Conclusions Laparoscopic surgery is effective for endometriosis, but the danger of recurrence should not be overlooked.

Objective To explore the mechanism action of Shoutai Pill in the embryo from the molecular level. Methods The model of normal pregnancy was established with the model of recurrent abortion CBA/J ×DBA/2. The recurrent abortion model CBA/J×DBA/2 in pregnant mice were randomly divided into model group, high-, medium-, low-dose group of Shoutai Pill. From the first day of gestation, mice were given medicine by gavage for 14 d, and then sacrificed. Immunohistochemistry was used to detect differences in protein HSP27,α-enolase, transferrin, annexin A2 protein expression. Results Compared with normal group, decidual HSP27 and α-enolase expression of model group increased significantly, the expression of transferrin and annexin A2 was significantly decreased, with significant differences (P<0.01). Compared with model group, decidual HSP27 andα-enolase expression of Shoutai Pill high and low dose group decreased, annexin A2 expression was increased. Compared with model group, expression of decidual transferrin of Shoutai Pill high dose group increased, with significant differences (P<0.01), decidual transferrin expression of Shoutai Pill medium and low dose group had no significant difference (P>0.05). Conclusion Through the protein expression, Shoutai Pill achieves the maintenance of pregnancy, reducing the rate of embryo resorption, which may be one of the mechanisms of Shoutai Pill preventing miscarriage effect.