Objective To explore the effects of alcoholic extracts of traditional Chinese medicines on the post-translational processing and trafficking of tyrosinase.Methods Human YUGEN8 amelanotic melanoma cells were grown in vitro;the cells were incubated with one of the seven traditional Chinese medicines,including Rhizoma Chuanxiong and psoralen.Protein analysis with Western blot,enzymolysis with endoglycosidase H (Endo H),and subcellular localization with laser confocal microscopy were per- formed.The expression,maturity and export from endoplasmic reticulum (ER) of tyrosinase in the treated cells were compared with those in the untreated controls.Results Compared with controls,an approximate- ly 80-kDa,Endo H-resistant tyrosinase doublet,which represented mature glycoform of tyrosinase,was in- creased in melanocytes treated with Semen Cuscutae,and in those treated with Semen Persicae.Within those cells,tyrosinase was distributed outside ER resident protein calnexin.Conclusion Both Semen Cus- cutae and Semen Persicae could induce tyrosinase maturation,stability and export from ER to distal site.

Cardiomyopathy, Dilated ; etiology ; genetics ; Humans

Objective To discuss the technical points and clinical effects of thoracic reconstruction after resection of the sternal tumnors with autogenous rib graft and pedicle oinental flap. Method A retrospective study was made on 12 patients,who underwent resection of sternal tumors and thoracic reconstruction with autogenous rib graft and pedicle omental flap between January 2004 and March 2010. The tumor involved the manubrium in 4 patients and the sternal body in 8 patients. Results All operations of 12 patients were succeed with no perioperative mortality or severe complications. Incisions healed by primary intention healing,and no paradoxical breathing was found. The postoperative period was uneventful.Conclusion It's an effective way to treat sternal tumors by the resection and thoracic reconstruction with autogenous rib graft and pedicle omental flap.

OBJECTIVEThe biological safety of a new developed silicone rubber for inflatable silastic prosthesis (SRISP) was evaluated.

METHODSFollowing the GB/T 16886.10-2005 standard, YY/T 0127.13-2009 standard, and GB/T 16886.11- 2011 standard, samples were prepared and tested by animal experiments, such as guinea pig maximization test, oral mucous membrane irritation test, and short-term systemic toxicity test (oral route).

RESULTSNo obvious erythema and edema in the guinea pig abdominal skin were observed after 24, 48, and 72 h of stimulating touch, thus indicating that SRISP does not cause potential skin sensitivity. No local response to SRISP was found, and the visual observation and pathological findings of oral mucosa were normal and similar to that of the control group. Therefore, SRISP had no irritation response to oral mucosa. No clinical signs of toxicity were observed in rats, and no significant differences in weight and weight relative growth rate between extract group and blank control group (P > 0.05) were found. Thus, SRISP had no short-term systemic toxicity.

CONCLUSIONThese results indicated that SRISP met the requirement of biomedical materials and had good bio- security.

Animals ; Biocompatible Materials ; Cosmetics ; Dimethylpolysiloxanes ; Guinea Pigs ; Prostheses and Implants ; Rats ; Silicone Elastomers ; Toxicity Tests

This study evaluated the cytotoxicity of a new type silicone rubber for maxillofacial prosthesis, which was developed by the present authors. According to the GB/T16886. 5- 2003, the samples were prepared and tested with cell counting kit-8 (CCK-8) assay, the relative growth rate (RGR) was calculated, and morphology of L929 cells were observed by scanning electron microscope and phase contrast microscope. The results showed that RGR of L929 cells were 91.65% (24 h), 87.03% (48 h), 87.30% (72 h), respectively, and the level of cytotoxicity was grade 1. The L929 cells showed typical fusiform shape and their morphology did not changed significantly after 24 h, 48 h and 72 h. These data indicated that the newly-developed silicone rubber material, as a maxillofacial prosthesis material, should be a safe biomaterial.
Animals ; Biocompatible Materials ; Cell Line ; Humans ; Maxillofacial Prosthesis ; Mice ; Silicone Elastomers ; toxicity

OBJECTIVETo determine the different rates of human papillomavirus types 6 (HPV-6) and 11 (HPV-11) infection in biopsy samples from pointed condyloma patients, and to construct prokaryotic expression system of the major capsid protein L1 of the virus so as to establish an ELISA for detecting the expression of L1 gene in the biopsy samples.

METHODSUsing a double PCR based on the L1 gene of HPV-6 and HPV-11, the infection rates of HPV-6 and HPV-11 in the biopsy samples were determined. The whole length of HPV-6 L1 gene was amplified using PCR and the target amplification fragment was sequenced after T-A cloning. The prokaryotic expression system pET32a-L1-E. coli BL21 (DE3) was constructed and SDS-PAGE was used to measure the expression of the target recombinant protein rL1. Rabbit anti-rL1 serum was prepared and immuno-diffusion assay was applied to examine the antiserum titer. ELISA was established to detect the expression of L1 gene in the biopsy samples.

RESULTSIn the biopsy samples from 116 pointed condyloma patients, 92.2% (107/116) were detectable for HPV-6 and/or HPV-11. Of the 107 positive samples, 70.1% (75/107) and 23.4% (25/107) were positive for HPV-6 or HPV-11 alone and 6.5% (7/107) were coinfected with both HPV-6 and HPV-11 respectively. When compared with the reported corresponding sequences, the homology of nucleotide and sequence of the cloned HPV-6 L1 gene was from 99.20% - 99.93% while its putative amino acid sequence homology was from 99.80% - 100%, suggesting IPTG could induce the expression of rL1. The immuno-diffusion titer of the rabbit anti-rL1 serum was 1:4. 88.8% (103/116) of the biopsy samples were the major capsid protein L1 detectable.

CONCLUSIONA prokaryotic expression system of HPV-6 L1 gene, a double PCR assay for HPV-6 and HPV-11 genotyping, and an ELISA assay for detecting the major capsid protein L1 were successfully established in this study. The pointed condyloma patients in Zhejiang area mainly infected with HPV-6. The HPV in the focus frequently expressed major capsid protein L1.

Animals ; Biopsy ; Capsid Proteins ; genetics ; Condylomata Acuminata ; pathology ; virology ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Human papillomavirus 11 ; genetics ; isolation & purification ; Human papillomavirus 6 ; genetics ; isolation & purification ; Humans ; Papillomavirus Infections ; virology ; Polymerase Chain Reaction ; Rabbits ; Sequence Homology, Nucleic Acid

AIM: To study the relation ship between stress and lung injury caused by acute hemorrhagic necrotizing pancreatitis(AHNP) through AHNP model.METHODS: The AHNP model was made by using 5% sodium taurocholate retrograded injection into biliopancreatic duct in SD rats. Those rats were divided into three groups randomly, from A to C, the A group undertook sham operations, the B group was made into AHNP model, and the C group was given Metyrapone. The level of corticosteroid, CRP and amylase in serum had been observed. The lung and pancreas histological examinations were also performed.RESULTS: In C group, the level of corticosteroid, CRP and amylase were much lower than those in B group. The grade of lung and pancreas injury were also lower than those in B group(P

Objective To construct a fusion gene (h1a-spaO) encoding H1a-SpaO protein of Sal-monella paratyphi A ( S.paratyphi A) and to express it in prokaryotic expression system , then to further ana-lyze the immunoprotective effects of the expressed protein rH 1a-SpaO.Methods The h1a-spaO fusion gene formed from separate h1a and spaO genes was amplified by PCR using flexible peptide sequence-containing linking primers and then sequenced after T-A cloning.A prokaryotic expression system for expressing h1a-spaO fusion gene was constructed by using the genetic engineering technique .The expressed protein rH1a-SpaO was examined by SDS-PAGE.The antigenicity and immunoreactivity of rH1a-SpaO protein were deter-mined by Western blot assay .The ability of rH1a-SpaO antiserum agglutinating S.paratyphi A strains was detected by micro-Widal′s test.The immunoprotective effects of rH 1a-SpaO against the lethal dose challenge of S.paratyphi A strains were analyzed in a mouse model and that were compared with those by using equal dose of individual recombinant protein H1a and SpaO (rH1a and rSpaO) as the immunogens, respectively. Results The h1a-spaO fusion gene was 100%identical with the individual h1a or spaO gene in nucleotide and amino acid sequences .The constructed prokaryotic expression system could express the recombinant pro-tein rH1a-SpaO with an advantage of high efficiency .rH1a-SpaO protein was able to react with rH 1a or rSpaO antiserum.Moreover, rH1a-SpaO antiserum also could efficiently recognize rH 1a and rSpaO as well as agglutinate Salmonella paratyphi A strains by binding with H-antigen.The immunoprotective rate (93.3%) in mice pre-immunized with 100 μg of rH1a-SpaO protein was significantly higher than that in those pre-immunized with equal dose of rH1a (60.0%) protein or rSpaO protein(53.3%) (P<0.05).Conclusion The recombinant fusion protein rH 1a-SpaO showed more stronger immunoprotective function than the individ-ual rH1a or rSpaO protein , which could be used as an effective antigen for the development of bi -valent para-typhoid A vaccine or typhoid/paratyphoid capsular polysaccharide-protein combined vaccine .

ObjectiveTo summarize the initial experience of transumbilical laparoendoscopic single-site surgery of urology.MethodsFrom February 2010 to March 2011,21 patients underwent laparoendoscopic single-site surgery using transumbilical single-site and common surgical instruments of laparoendoscopic.Nine patients underwent single-site laparoscopic ureterolithotomy,5 underwent transumbilical single-site laparoscopic ureteral stricture resection and anastomosis,5 underwent transumbilical single-site laparoscopic renalcyst unroofing and 2 had a nephrectomy.All of the cases were definitely diagnosed.A single umbilical incision of 1.5 cm to 2.5 cm was made for Triport.The procedures were performed according to the methods used in classical laparoscope methods using general instruments.ResultsAll the operations were successfully completed without conversion to open surgery.The mean operative time of ureterolithotomy was 143 (120-230) min,the mean operative time of ureteral stricture resection and anastomosis was 157 (120 -180) min,the mean operative time of unroofing of renal cysts was 110 (95 -132) min,and the operative time of the nephrectomy was from 95 to 120 min.The intestinal tract function recovered within 1 -2 d,the drainage tube was removed within 2 -3 d and the postoperative hospitalization duration was 4 -7 d.The symptoms were reduced or disappeared and no major intraoperative or postoperative complications occurred within 4 - 6 months.Conclusions Transumbilical laparoendoscopic single-site surgery represents a safe and feasible operation for urologic patients.With more clinical practice,laparoendoscopic single-site surgery could be generally applied.

Objective To investigate the prevalence rate of healthcare-associated infection(HAI)in a hospital,so as to provide reference for making HAI control measures.Methods The cross-sectional survey on HAI was carried out among all hospitalized patients on May 26,2010,December 12,2012 and December 4,2013,respectively,sur-veyed data were analyzed.Results The prevalence rate was 6.66%(n =116),6.67%(n =113)and 6.33%(n =120)in 2010,2012 and 2013 respectively,and case rate was 7.29%(n=127),7.38%(n=125)and 6.97%(n=132) respectively,intensive care unit(ICU )had the highest infection rate,internal medicine ICU was up to 71 .43%. The main infection site was lower respiratory tract(44.53%),followed by surgical site infection (9.11 %)and uri-nary tract infection (9.11 %).The isolation rate of gram-negative bacteria,gram-positive bacteria and fungi was 60.81 %,20.38% and 18.81 % respectively.Usage rate of antimicrobial agents in three years was 32.95%, 29.87% and 25.59% respectively (χ2 = 13.16,P <0.01 ).Conclusion Prevalence rate of HAI in this hospital is high ,the main pathogen is gram-negative bacteria,the main infection site is lower respiratory tract ,antimicrobial use decreased year by year.Monitor on high risk departments,main sites and pathogens should be intensified.