Objective To prepare B-lymphoblastoid cell lines of HLA novel allele B*5610 in a family for further study and identification . Method Isolate mononuclear cells under aseptic conditions from the peripheral blood. After infection with Epstein-Barr virus, the cells were cultured in 20% FBS, 2?g/ml CsA RPMI 1640. Results Immortalized B-lymphoblastoid cell lines of five B *5610 carriers in a family were achieved, and the new genes were inherited stably. Conclusion Our work is important for storing and breeding the precious material of biomedicine because the B *5610 genes in the immortalized B-lymphoblastoid cell lines were inherited stably.

Biodiesel, a nontoxic,cleaning, renewable and biodegradable fuel, is expected as a substitute for conventional fossil diesel. There are three main approaches to produce biodiesel, alkali-catalysis processing, enzymatic-catalysis processing and supercritical processing. With the unique property of energy-saving and environment-friendly, enzymatic-catalysis appears a great potential for industrial application. The main bottleneck of this technology is high cost and low stability of the lipase, as well as the inactivation of lipase by methanol and so on. To settle the problem, several methods have been used including the fixed-bed bioreactor, enzyme immobilized processing, whole-cell biocatalyst, changing addition method of methanol, developing of novel acyl acceptor, enhancing methanol resistance of lipase. The main problems and the relative strategy research of the enzymatic-catalysis technology were sum up.

Lipase gene from Penicillium expansum(lip07) was cloned and over-expressed in Pichia pastoris.a random mutant named ep8,which contained a single amino acid substitution,was obtained by using the lip07 as an error-prone PCR template in previous study.ep8 shows higher thermostability than that of lip07,To further improve the thermostability of the lipase,the Lys of wild-type(lip07) and mutant(ep8) in 202 were substituted by Ala using the Overlap extension PCR technique respectively.The mutant genes(lip07-K202A and ep8-K202A) were subcloned into pAO815,and then transformed into the Pichia pastoris GS115 for extracelluar expression,respectively.15% SDS-PAGE analysis indicated that the molecular mass of PEL-ep8-K202A and PEL-lip07-K202A are both about 28kDa,which is same with the wild-type lipase.The Tm of PEL-ep8-K202A is 41.66℃,2.63℃ higher than that of the wild-type(39.03℃) and 1.21℃ higher than the random mutant(PEL-ep8:40.45℃);the Tm of single mutant(PEL-lip07-K202A) is 37.08℃,2℃ lower than that of the wild-type lipase.

Objective To explore the relationship of expression and the effect of preoperative radiotherapy of endothelin-1 (ET-1) and pyruvate kinase M-2 (PKM2) in rectal carcinoma.Methods Immunohistochemical method was used to detect the expression of ET-1 and PKM2 proteins of rectal cancer tissues in 96 cases.The expressions of ET-1 and PKM2 were analyzed with the effect of preoperative radiotherapy in rectal cancer tissue.Results The high expression of ET-1 protein was 59 cases (61.46％).The high expression of PKM2 proteins was 54 cases (56.25％).The high expressions of ET-1 and PKM2 protein were worsen the effect of tumor regressive grade (TRG) than lower expressions of those after preoperative radiotherapy of rectal cancer tissue (P ＜ 0.05).The protein expression of ET-1 and PKM2 were positively correlated (P =0.006).Conclusions The high expressed ET-1 and PKM2 proteins in rectal cancer are closely related to preoperative radiotherapy resistance.ET-1 and PKM2 proteins are expected to become new targets of radiotherapy sensitivity and radiotherapy sensitization of rectal cancer.

Objective To study the effects of rhubarb on the mitogen-activated protein kinase (MAPK) sig-naling transducfon pathway in rats with severe acute pancreatitis (SAP),and to investigate the treatment mecha-nism of rhubarb on SAP. Method One hundred SD rats were provided by from the Animal Center of Nanjing Uni-versity. All animals were randomly divided into sham operation (n=33), SAP (n=33) and rhubarb groups (n=34). SAP model was induced by retrograde injection of 5% sodittm taurocholate. Rhubarb was given with 10% rhubarb decoction (2 mi/100 g) at the time of pancreafitis induction in the rhubarb groups. At 1, 3, 6, and 12 h after the models were established,animals were killed. MAPK activity in pancreatic tissue was examined by West-em blotting and the mRNA levels of TNF-α and IL-6 in pancreatic tissues were detected by RT-PCR. All data were analyzed by SPSS statistical software and statistical differences between values from two sroups were determined by the Student's t -test. Results MAPK activity, TNF-α and IL-6 mRNA levels in pancreatic tissues were signifi-cantly enhanced in the SAP group compared with the sham operation group (all P<0.01). Rhubarb treatment markedlyinhibited MAPK activation,TNF-α,IL 6 mRNA (all p<0.01). Conclusions Rhubarb can alleviate the inflammatory response of SAP by down-regulating MAPK activity.

Background and purpose: The incidence and mortality of lung cancer ranked first in China. This study aimed to describe lung cancer survival in Shanghai, and provide background information for cancer prevention and treatment evaluation. Methods: Data of lung cancer cases diagnosed during 2002-2006, follow-up information and death report were collected from Shanghai Cancer Registry. Life table method and Ederer Ⅱ were used to calculate observed survival (OS) and relative survival (RS) respectively. Related demographic characteristics and status were also analyzed to present the survival situations of the lung cancer survivors in Shanghai. Results: In this study, 41802 lung cancer cases were included in analysis. The 5-year OS and RS for lung cancer were 13.75% and 20.23% respectively, and median survival time was 318 days. Survival rate was higher among females than males, with the 5-year OS of 15.49% and 13.00% respectively. The 5-year OS was higher among suburban residents (14.25%) than urban residents (13.23%). Survival rates decreased with increasing age and advanced stage. Patients aged 0-34 had a 5-year OS of 38.21%, while patients aged above 75 had a 5-year OS of 5.48%. Patients diagnosed with stage Ⅰ had a 5-year OS of 55.47%, while patients diagnosed with stage Ⅳ had a 5-year OS of 5.27%. Survival of lung cancer patients differed by tumor histological subtype. The 5-year OS of squamous lung cancer (24.40%) was higher than other histological types, followed by adenocarcinoma (22.26%), large cell (20.27%) and small cell lung cancer (12.22%). From 1972-1976 to 2002-2006, the 5-year OS of urban male patients increased from 6.8% to 12.4%, and 5-year OS of urban female patients increased from 7.3% to 14.9%. Analysis of RS gave the similar results. Conclusion: During the past 30 years, survival rate of lung cancer patients in Shanghai improved steadily, and the survival condition is above average among different countries and areas. However, survival rate of lung cancer is still low compared with other types of cancer. Future focus should be placed on the control of tobacco smoking, early detection by low-dose helical computed tomography and targeted therapy to further improve lung cancer survival.

OBJECTIVETo detect the expression of human telomerase reverse transcriptase (hTERT) protein and mRNA in bile duct carcinomas and the adjacent tissues and to elucidate its role in bile duct carcinogenesis.

METHODSThe expression of hTERT protein and hTERT mRNA in the formalin-fixed paraffin-embedded specimens of 71 cases of bile duct cancers and 39 cases of adjacent tissues was detected by streptavidin-peroxidase immunostaining and in situ hybridization. The correlation was analysed statistically between the expression of hTERT protein and mRNA and clinicopathological parameters bile duct carcinomas.

RESULTSThe positive rate of hTERT protein expression and mRNA expression in malignant specimens was 78.9% (56/71) and 67.6% (48/71), while that in the adjacent tissues was 35.9% (14/39) and 23.1% (9/39), respectively. All the positive signals were found in the hyperplastic biliary epithelia. No significant correlation was established between hTERT expression and clinicopathological parameters.

CONCLUSIONhTERT gene transcription and protein expression is most likely involved in the proliferation and malignant transformation of bile epithelia and the malignant progression of bile duct carcinomas. The detection of hTERT expression may serve elucidating the carcinogenesis of bile duct.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Neoplasms ; enzymology ; pathology ; DNA-Binding Proteins ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; RNA, Messenger ; analysis ; Telomerase ; analysis ; genetics

OBJECTIVETo investigate the dynamic expressions of TGF-beta 1 and TGF-beta 1 mRNA at different stages of hepatocellular carcinoma (HCC) development and their use in clinical diagnosis.

METHODSHepatoma models were developed with 2-FAA using male Sprague-Dawley (SD) rats. Morphological changes of the rat liver histological preparations (H and E stained) were studied. The fragment of TGF-beta 1 gene obtained was amplified by nested RT-PCR. Dynamic change of TGF-beta 1 level was quantitatively analyzed by ELISA. The distribution of TGF-beta 1 in the cells and its gene expression were detected in human HCC tissues.

RESULTSThe progressive increases of hepatic TGF-beta 1 and TGF-beta 1 mRNA were observed in rat hepatocytes which progressed from granular degeneration, atypical hyperplasia and finally to HCC development induced by 2-FAA. The expression levels in HCC tissues were significantly higher than those in the normal and degenerative ones. TGF-beta 1 was shown in rat hepatocytes by immunohistochemistry. Plasma TGF-beta 1 was detected in 89.5% of all the patients with HCC, but it was detected in 93.3% of them who had an AFP less than 400 microg/L. TGF-beta 1 mRNA showed a stronger expression in HCC tissues. TGF-beta 1 mRNA was found in peripheral blood mononuclear cells from all HCC patients with extrahepatic metastasis.

CONCLUSIONTGF-beta 1 may participate in hepatocyte canceration. The overexpression of TGF-beta 1 and TGF-beta 1 mRNA could be useful markers for early diagnosis and predicting prognosis of HCC.

Adult ; Aged ; Animals ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; blood ; diagnosis ; pathology ; Female ; Humans ; Liver Neoplasms, Experimental ; blood ; diagnosis ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; prevention & control ; Prognosis ; RNA, Messenger ; blood ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; blood

This study aimed to examine the biocompatibility of calcium titanate (CaTiO3) coating prepared by a simplified technique in an attempt to assess the potential of CaTiO3 coating as an alternative to current implant coating materials.CaTiO3-coated titanium screws were implanted with hydroxyapatite (HA)-coated or uncoated titanium screws into medial and lateral femoral condyles of 48 New Zealand white rabbits.Imaging,histomorphometric and biomechanical analyses were employed to evaluate the osseointegration and biocompatibility 12 weeks after the implantation.Histology and scanning electron microscopy revealed that bone tissues surrounding the screws coated with CaTiO3 were fully regenerated and they were also.well integrated with the screws.An interfacial fibrous membrane layer,which was found in the HA coating group,was not noticeable between the bone tissues and CaTiO3-coated screws.X-ray imaging analysis showed in the CaTiO3 coating group,there was a dense and tight binding between implants and the bone tissues;no radiation translucent zone was found surrounding the implants as well as no detachment of the coating and femoral condyle fracture.In contrast,uncoated screws exhibited a fibrous membrane layer,as evidenced by the detection of a radiation translucent zone between the implants and the bone tissues.Additionally,biomechanical testing revealed that the binding strength of CaTiO3 coating with bone tissues was significantly higher than that of uncoated titanium screws,and was comparable to that of HA coating.The study demonstrated that CaTiO3 coating in situ to titanium screws possesses great biocompatibility and osseointegration comparable to HA coating.

OBJECTIVETo study the durability of expanded polytetrafluoroethylene artificial heart valve (ePTFE AHV).

METHODSSix ePTFE AHVs were tested for 400 million times against accelerated fatigue using TH-2200 artificial heart valve exosomatic accelerated fatigue instrument. Hydromechanical parameters of fore-and-aft accelerated fatigue test of the 6 AHVs were obtained by TH-1200 artificial heart valve exosomatic pulsatile stream instrument.

RESULTS AND CONCLUSIONThe mean gradient pressure spanning the valve and the effective orific area of ePTFE AHVs did not undergo significant changes after fore-and-aft the fatigue test, but the regurgitation volume and regurgitation rate of ePTFE AHVs were reduced after the accelerated fatigue test, suggesting good durability of ePTFE AHV.

Biocompatible Materials ; chemistry ; Heart Valve Prosthesis ; standards ; Materials Testing ; methods ; Polytetrafluoroethylene ; chemistry ; Tensile Strength