Objective A method was established for genotyping of hepatitis B virus (HBV),based on the restriction fragment length polymorphism(RFLP)created by BsrI, StyI,DpnI and HpaII action on an amplified segment of the S region. Methods 223 full-genomic sequences were analyzed and the aligned nucleotide and amino acid sequences of S gene, genotype specific regions were identified by the restriction enzymes, BsrI, StyI,DpnI and HpaII. Pre S PCR-RFLP genotyping method was applied to a number of serum samples from hepatitis B e antigen (HBeAg) positive and negative Chinese chronic HBV carriers. And in 90 samples the following genotypes were observed: 30B, 30 C, 30D. The method using S gene PCR-RFLP was confirmed to be correct by these 90 samples. Three samples of each genotype B, C and D were randomly selected and directly sequenced their S gene to confirm that HBV S gene PCR-RFLP genotyping method was correct disectly. Results The results of two PCR-RFLP HBV genotyping methods were coincide with that of S gene sequence. Conclusions The method for genotyping of hepatitis B virus (HBV), based on S gene RFLP is established to be highly sensitive, differential and accurate. The RFLP patterns are easy to be recognized because of its simplicity and singleness.

This study is to investigate the protein and mRNA expressions of pro-inflammatory and anti-inflammatory cytokines in U937 foam cells and effects of Ginkgo biloba extract (GbE) on the cytokines.U937 cells were cultured with different concentrations of GbE (0.1,1,and 10 μg·L-1),and stimulated by 100 mg·L-1 oxidized low density lipoprotein (ox-LDL) for 24 h.The expressions of interleukin-1β (IL-1β),tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in culture solution were detected by enzyme-linked immunosorbant assay (ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR).The results showed that incubated with 100 mg·L-1 ox-LDL for 24 h,the U937 cells became foam cells,the protein or mRNA expressions of IL-1β,TNF-α,IL-10,and its receptor IL-10R in U937 foam cells were higher markedly than those in normal U937 cells.When the cells were pretreated with GbE (0.1,1,and 10 μg·L-1),the increases of IL-1β and TNF-α in U937 foam cells were remarkably inhibited,but IL-10 expression increased greatly.Especially when cells were pretreated with 10 μg·L-1 GbE,the protein and mRNA expressions of IL-1β and TNF-α were markedly lower than those in U937 foam cells.The protein expression of IL-10 and mRNA expressions of IL-10 and its receptor IL-10R were markedly higher than those in U937 foam cells.GbE inhibited production of pro-inflammatory cytokines IL-1β and TNF-α,but up-regulated the production of anti-inflammatory cytokine IL-10 and its receptor IL-10R in U937 foam cells,which might be related with its anti-atherosclerotic actions.

Objective:To evaluate the short-term effect of robotic pancreaticoduodenectomy (RPD) and laparoscopic pancreaticoduodenectomy (LPD) in the treatment of ampullary and periampullary tumor systematically.Methods:A systematic search of the PubMed, Embase and Cochrane library database using the key words "pancreaticoduodenectomy, duodenopanreatectpmy, whipple, laparoscopic, robotic, Da Vinci" . A systematic search of the Sinomed, Wangfang, VIP and CNKI databases including the key words "胰十二指肠切除" , "腹腔镜" , "机器人" , "达芬奇" . To investigate the differences of the conversion rate of laparotomy, incidence of postoperative complications, and the postoperative hospital stay between the RPD group and the LPD group. The software RevMan5.3 was used in this meta-analysis.Results:A total of 4 retrospective cohort studies and 1 001 patients were included in this meta-analysis, including 451 patients in the RPD group and 550 patients in the LPD group. The results of the meta-analysis showed that there were significant differences between RPD group and LPD group for the conversion rate laparotomy ( OR=0.35, 95% CI: 0.24-0.50, P<0.05). There were no significant differences between RPD group and LPD group for the incidence rate of overall complication( OR=1.23, 95% CI: 0.95-1.58, P>0.05), hemorrhage ( OR=0.71, 95% CI: 0.50-1.00, P>0.05), pancreatic fistula ( OR=1.09, 95% CI: 0.80-1.49, P>0.05), delayed gastric empty ( OR=0.81, 95%CI: 0.57-1.14, P>0.05) and hospital stay after surgery ( WMD=-2.87, 95% CI: -1.44-1.70, P>0.05). Conclusions:RPD is as safe as LPD, with the same complication rate. Therefor it is worthy for further application in medical institutions with relevant conditions.

AIM: To investigate the differentiative and apoptotic effect of CpG-oligodeoxynucleotides on HL60 cells and its mechanism. METHODS: After HL60 cells were exposed to synthetic CpG-oligodeoxynucleotides, non-CpG-oligodeoxynucleotides or ZpG-oligodeoxynucleotides for 72 hours, respectively, the inhibition of HL60 cells were detected using MTT method, NBT test was used and CD14 expression were determined. Apoptosis of HL60 cells were mensurated with flow cytometry and transmission electron microscope, and caspase 3, Bcl-2 and Bax expression of HL60 cells treated with oligodeoxynucletides were detected using immunohistochemistry. RESULTS: Treatment with CpG-oligodeoxynucleotides induced the differentiation and apoptosis in HL60 cells, but non-CpG-oligodeoxynu- cleotide and ZpG-oligodeoxynucleotide had no effect on HL60 cells. CONCLUSION: CpG-oligodeoxynucleotides can induce the differentiation and apoptosis in HL60 cells. It may provide a new approach for the immunological treatment of leukemia.

Background Keratectasia after laser in situ keratomileusis (LASIK) is a rare but severe complication,which threatens the visual acuity and corneal strength.Corneal collagen crosslinking (CXL) is a new therapy that increases the security and decreases the risk of complication.However,the effectiveness and safety of LASIK-CXL is still need to be concerned.Objective This study was to evaluate the safety of LASIK-CXL for myopia and astigmatism with thin cornea.Methods A prospective cohort study was designed.A total of 128 eyes of 64 patients with thin corneal and myopic astigmatism enrolled in Beijing Tongren Eye Center from January 2014 to January 2015.The patients were assigned to LASIK group (74 eyes of 37 patients) and LASIK-CXL group (54 eyes of 27 patients).Refractive surgery was performed by Visumax femtosecond lasrer and VISX S4 excimer laser.Eyes of LASIK-CXL group applied accelerated CXL immediately after LASIK.The follow-up was 6 months.Manifest refraction,uncorrected (UDVA) and corrected distance visual acuity (CDVA),average keratometry values (AveK),anterior segment OCT (AS-OCT),corneal hysteresis (CH) and corneal resistance factor (CRF) were examined before and after operation.This research passed through Ethics Committee of Beijing Tongren Hospital.Results The spherical equivalent (SE) of the LASIK group and LASIK-CXL group were (-6.49 ±2.41)D and (-6.97 ±2.41) D before operation and decreased to (-0.68 ±0.88) D and (-0.75 ±0.94) D 6 months after operation.The UDVA (LogMAR) was 1.18±0.28 and 1.05±0.38 before operation and elevated to-0.06±0.09 and-0.03±0.186 months after operation in the LASIK group and LASIK-CXL group.The preoperative AveK values were (44.37 ±1.46) D and (44.47± 1.50)D in the LASIK group and LASIK-CXL group and reduced to postoperative (39.30±2.06) D and (38.66± 1.80) D.The preoperative SRI of LASIK group and LASIK-CXL group were 0.25 ±0.21 and 0.24±0.22,which increased to 0.29±0.24 and 0.28±0.24.The SAI values were 0.36±0.16 and 0.39±0.15 before operation,which increased to 0.57 ±0.31 and 0.75 ±0.376 months after operation,and the SAI value of the LASIK-CXL group was significantly higher than that of LASIK (F =10.220,P--0.002).CRF values of LASIK and LASIK-CXL were (8.44±1.44)mmHg and (8.63±1.35) mmHg in preoperation,which decreased to (5.74±1.31) mmHg and (6.25± 1.24) mmHg in postoperation.The result of LASIK-CXL was higher than that of LASIK (F=8.650,P =0.040).CH values were 8.78 ± 1.51 and 8.69 ± 1.62 in preoperation,which decreased to (7.23 ± 1.08) mmHg and (6.50±1.32)mmHg.The value of LASIK-CXL was lower than that of LASIK (F =5.860,P =0.017).The mean depth of demarcation line was (228.45±28.24) μm (range 165 to 310 μm) on OCT,which was presented in 45 eyes (81.82％) at 1 month in postoperation.Conclusions Accelerated CXL with FS-LASIK is effective and safe in improving visual acuity in myopic astigmatism patients with thin cornea,which also can increase the rigidity of the cornea.

The experiment was conducted between February 2004 and May 2006 at The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang province, China. Forty hybrid pigs were used in the experiment, 20 were used as donors and the other 20 were used as recipients. Donor operation: Abdominal aorta was lavaged in situ with 500 mL 4℃ heparinized saline (lavage pressure 7.8-9.8 kPa), intestinal canal cut from donor was conserved in 4℃ physiological saline.Recipient operation: Orthotopic small intestine transplantation and heterotopic small intestine transplantation were conducted according to exploring results, and abdominal cavity was washed with 40-45℃ physiological saline until the flush out liquid became warm.Periprocedural management: Preoperative discussion and postoperative discussion were done, and body temperature of pigs was monitoring and controlled. Early-stage feeding was used, 4 pigs could drink water freely after transplantation and took food normally at the second day, while other 16 pigs began to drink water 2 days after transplantation. Evaluation after the operation: No experimental animal had anastomotic leakage after taking food. Sixteen pigs survived over 7 days, average 9 days, the longest over 14 days. The success rate of the small intestine transplantation is 80% (16/20). Failure reasons: One with phlebothrombesis, one with blood loss and two with unknown reasons. Results showed that experience of periprocedural management had got relatively satisfactory effects through a great quantity of small intestine transplantation practices.

Objective To investigate the feasibility and clinical effect of immediate ureterovesical reimplantation after ureteral rupture during laparoscopic hysterectomy. Methods From August 2010 to December 2015, 5 cases of ureteral rupture during the operation of laparoscopic hysterectomy were treated with immediate ureterovesical reimplantation under laparoscopy. Results All operations were successfully performed without transversion to open surgery. No patients with urinary leakage occurred. The mean follow-up were 21 months (range 3-60 months) . No cases with ureteral stricture were observed. Slight urine reflux was found in two patients, of whom obvious hydronephrosis and renal damage were not found. Conclusion Immediate ureterovesical reimplantation under laparoscopy is a feasible, safe and minimal invasive method for treatment of ureteral ruputure during laparoscopic hysterectomy.

Abdominal Pain ; etiology ; Acute Pain ; etiology ; Child ; Humans ; Lymphoma ; complications

Objective To investigate the clinical characteristics,diagnosis and treatment of congenital tracheobiliary fistula (CTBF) in children.Methods A case of CTBF admitted into the Department of Pediatric Respiration,Hubei Maternal and Child Health Care Hospital in 2016 was reported,and the related literatures were reviewed.The clinical features,diagnostic methods,treatment status and clinical outcomes of the disease were analyzed.Results The patient was 3 years and 7 months old.The main clinical manifestations were recurrent cough,pneumonia and atelectasis.CTBF was diagnosed by means of iodine oil radiography and treated with biological glue plugging under bronchoscopic guidance.So far,only 30 cases of CTBF have been reported in the English literatures,but only 2 cases in the Chinese literatures.The main clinical manifestations were cough,dyspnea,sputum or bile vomiting,aspiration pneumonia,atelectasis or emphysema.Diagnostic methods for CTBF included bronchoscopy,bronchial angiography and cholangiography,hepatobiliary scan,CT scan and magnetic resonance imaging.Except for this case treated with biological glue plugging under bronchoscopic guidance,all other patients were treated with surgery.The operation methods included fistula ligation,gastrostomy,liver resection,fistula jejunum Roux-en-Y anastomosis,hepatic hilum jejunum anastomosis,gallbladder jejunum anastomosis,etc.Only 4 cases died,and the rest of the patients recovered.Conclusions CTBF should be suspected in children with persistent chronic cough.Iodine oil radiography through bronchoscopy is a simple and feasible method for diagnosis of CTBF.Besides surgery,the lavage and the biological glue plugging method through bronchoscopy is also an effective way to treat CTBF in children without severe biliary malformation.