Objective To construct the recombinant yeast expressing PreS2 120-146-hepatitis B surface antigen (HBsAg),and to evaluate the immune effects of whole yeast cells.Methods PreS2 120-146 and HBsAg gene sequence were optimized according to the yeast cell codon preference,and were recombined and cloned into pPIC3.5K yeast expression vector to construct pPIC3.5K/PreS2 120-146 plasmid.After digested and linearized by Bgk Ⅱ restriction enzyme,pPIC3.5K/PreS2 120-146-HBsAg recombinant plasmid was electrotransformed into GS115 strain to screen PreS2 120-146-HBsAg-recombinant Pichiapastoris .The expression of PreS2 120-146-HBsAg was identified by sodium doclecyl sulfate polyacrylamide gel electrophogesis (SDS-PAGE),Western blot and enzyme linked immunosorbent assay (ELISA)analysis. BALB/c mice were vaccinated by inactivated whole recombinant yeast cells expressing target protein. Specific antibodies to HBsAg were detected by ELISA.Cytotoxic T lymphocyte (CTL)response induced by interferon (IFN)-γ was detected by reverse transcription-polymerase chain reaction (RT-PCR)when immune spleen cells of mice were stimulated by CTL epitope on HBsAg.Independent sample t test was used. Results Data of PCR detection,restriction enzyme digestion and sequencing analysis showed that recombinant pPIC3.5K/PreS2 120-146-HBsAg plasmid was successfully constructed.SDS-PAGE,Western blot and ELISA verified the expression of PreS2 120-146-HBsAg in the lysate of the recombinant Pichiapastoris induced by methanol.Levels of specific anti-HBsAg IgG antibodies produced by inactivated yeast cells vaccinated mice were comparable to purified HBsAg immunization (t =0.946,P =0.381 ). Analysis of HBsAg-specific CTL responses revealed that the level of IFN-γwas significantly higher when the immune spleen cells of mice were stimulated by CTL epitope peptides on HBsAg (t =2.305 ,P =0.044).Conclusions PreS2 120-146-HBsAg target protein is successfully expressed by construction of recombinant Pichiapastoris . The specific humoral and cellular immune responses are induced by recombinant whole yeast cells vaccinated mice.

It is proven that laparoscopic sleeve gastrectomy (LSG) is effective for obesity, type 2 diabetes mellitus and other obesity-related complications. The surgical procedure of LSG is relatively simple, with few short-term or long-term complications, which has increasing be applicated in recent years. As more hospitals in China carry out LSG, the related reports of its complications have gradually increased. The most common complications of LSG include gastric leakage, bleeding, and gastric cavity stenosis. Among them, gastric leakage is a more difficult problem, and untreated or improper treatment can bring serious consequences to patients. Based on the current progress in clinical treatment of gastric leakage after LSG and practical experience, the authors summarize and discusse the clinical experience of the prevention and treatment of such patients.

Objective To evaluate the effectiveness of carboprost methylate suppository for cervical ripening before diagnostic hysteroscopy in premenopausal women. Methods From July 2014 to July 2015, 1614 women who were undergone diagnostic hysteroscopy in 12 hospitals were randomly assigned into study group(n=1209)and control group(n=405). The cases in study group were given 1 mg carboprost methylate suppository in vagina before hysteroscopy, the cases in control group were given 1 mg placebo. The extent of cervical ripening, the time of dilated cervix, pain scoring, incidence of drug side reactions after 24, 48, 72 hours, satisfaction degree of operators and patients, the time of hysteroscopy, incidence of complications between the two groups were observed and compared. Results (1) Mean cervical widths in the study and control groups were 6.11 ± 1.11 and 5.95 ± 1.11, and showed a significant difference(P=0.034);the percentage of women requiring cervical dilatation in study group was lower than the percentage in control group significantly [28.3％(342/1209)versus 34.6％(140/405), P=0.020].(2) The time of dilated cervix in study group was shorter than the time in control group significantly [(34 ± 25) versus(52 ± 49)s, P=0.028] for the patients whose mean cervical widths≤4.(3)There was no significant difference in pain scores between the two groups(P>0.05).(4)The incidence of side reactions 24, 48, 72 hours after operation were no significant difference between the two groups (P>0.05). (5) The satisfaction degree of operators and patients, the time of hysteroscopy, incidence of complications between the two groups were no singnifcant difference between the two groups (all P>0.05). Conclusion Application of carboprost methylate suppository by vagina before hysteroscopy is an effective and safe method of cervical ripening.

Objective: To evaluate the relationship between platelet distribution width(PDW) and the extent of coronary artery disease and 2-year outcome in patients received percutaneous coronary artery intervention(PCI) because of stable coronary artery disease(SCAD). Methods: We consecutively enrolled 4 293 patients who received PCI because of SCAD in Fuwai Hospital from Jan 2013 to Dec 2013, patients were followed up for 2 years. Patients were divided into three groups according to tertiles values of PDW as follows: PDW≤11.4%(1 402 patients), 11.4%12.9% (1 450 patients). Major adverse cardiovascular and cerebrovascular events (MACCE) were defined as the occurrence of death, myocardial infarction, target vessel revascularization, intra stent thrombosis and stroke during follow-up. Multivariable logistic regression was used to evaluate the relationship between PDW and the extent of CAD. Multivariable Cox regression was used to evaluate the relationship between PDW and prognosis of SCAD patients. Results: PDW was associated with diabetes mellitus, body mass index, red cell distribution width, mean platelet volume (MPV), platelet counts and glycosylated haemoglobin (P<0.05), but not associated with age, sex, estimated glomerular filtration rate (P>0.05). PDW was not correlated with the extent of CAD(P=0.990), SYNTAX score(P=0.721), no-reflow phenomenon after PCI(P=0.978). Multivariable logistic regression also showed no relationship between PDW and extent of CAD (OR=0.994, 95%CI 0.961-1.029, P=0.73). PDW was found to be an independent risk factor of 2-year cardiac death (HR=1.242, 95%CI 1.031-1.497, P=0.022), but was not an independent risk factor of all-cause death and MACCE. Conclusions PDW is not related with the extent of coronary artery disease. PDW is an independent risk factor of 2-year cardiac death, but is not an independent risk factor of all-cause death and MACCE in this patient cohort.