Objective: To highly express rh-Endostatin from Pichia pastoris and purify it to homogeneity. Methods: Constructed Pichia pastoris X33/pLW202 was amplified and inoculated to ferment media. The supernatant of the strain was collected after induction. Through ultrafiltration, affinity and gel chromatography, rh-Endostatin with high purication was obtained. Results: 60mg/L rh-Endostatin was obstained from supernatant. HPLC showed its purity was above 98%. Conclusion: High level expression of secreted rh-Endostatin has been successfully achieved in Pichia pastoris expression system.

Aim To explore inhibitory effects and mechanism of engineering endostatin on growth and metastasis of melanoma cells in mice. Methods Melanoma cells(2× 106/mouse)were inoculated sabcutaneously to C57BL/6 mice. After tumorigenesis,endostatin(8mg/kg.d)was administrated to tumor-bearing mice,once a day ,twenty-one in all.Dietetic state and weight change of the tumor-bearing mice were observed and tumorous sige was measured during administration of endostatin. On 26thday,the tumor-bearing mice were sacrificed,subcutaneously tumorous weight was weighed and brain,lung ,liver,spleen and kidney were excised and sections were made to supply the pathological examination. Results Area under curve in the endostatin-treated group was obviously less than that in tamor control group(P∨ 0.01). Pathological study revealed that lavge areal necrosis arose in tumor and newborn cappillaries around the tumor disapeared. Conclusion Endostatin possosses strongly inhibitory effects on growth and metastasis of mouse melanoma and formation of newborn capillaries around tumor.

Objective In order to extrapolate the respiratory dysfunction of patients in early stage of acute organophosphorus pesticide poisoning (AOPP),transpulmonary thermodilution technique was used in swine models of severe acute dichlorvos poisoning (SADP) to evaluate respiratory function.Methods Twenty healthy female swine were randomly divided into dichlorvos ( n =7 ),atropine ( n =7 ) and control (n =6) groups.In the dichlorvos group,the swine were administered with 80％ emulsified dichlorvos (100mg/kg) via the gastric tube toinduce SADP.In the atropine group,swinewere administered with dichlorvos,and 0.5h later,atropine was injected to obtain and maintain atropinization.The swine of control group were administered with saline solution instead.Arterial and venous blood samples were collected 0,0.5,1,2,4 and 6 hours after modeling for blood gas analysis and detecting acetylcholinesterase levels.Both extravascular lung water index (EVLWI) and pulmonary vascular permeability index ( PVPI ) were measured by using PiCCO (pulse indicator continuous cardiac output ). At the termination of the experiment,the animals were sacrificed and the lung wet/dry weight ratio was determined and histopathological changes of lung tissue were also observed under microscope.Results In the dichlorvos group,EVLWI and PVPI were substantially increased from 0.5 h to 6 h after modeling but PaO2/FiO2 decreased from 0-6 h after modeling.In the atropine group,EVLWI and PVPI increased initially,but then they decreased 1 h afterwards and PaO2/FiO2 was also gradually decreased from 0-1 h.In both dichlorvos group and atropine group,the EVLWI was negatively correlated with PO2/FiO2 and positively correlated with PVPI.Compared with the control group,the lung wet/dry weight ratio increased markedly in the dichlorvos group and mildly increased in the atropine group.Meanwhile,the histopathological changes of lung tissue were obvious in the dichlorvos group and mild in the atropine group.Conclusions SADP swine experienced substantial changes in respiratory function. EVLWI was a reliable and valuable indicator for evaluating respiratory function in the early stage of AOPP.

Objective To study the effects of laparoscopic and open hepatectomy(LH and OH) on liver function of patients,and to evaluate patients′ traumatic reaction after LH.Methods Forty patients scheduled to undergo hepatectomy were divided into LH group and OH group randomly in PLA General Hospital from Jun.2004 to Feb.2005.Data of pre-and post-operative liver function of the two groups were compared.Results Postoperative mean value of ALT,AST and LDH in LH group and OH group elevated quickly,and the postoperative mean value of these markers was significantly different compared to that of the preoperative value,and the mean value in OH was higher than that in LH.There was no significant difference between postoperative and preoperative level of ALP and ?-GT in both groups,but postoperative mean value of them in OH was higher than that in LH.There was no significant difference between the 2 groups in TBIL and DBIL level,but their postoperative mean value in OH was higher than that preoperatively.Conclusions The effects of LH on liver function in patients are less than those of OH.This finding suggests that patients′ traumatic reaction after LH is less than that after OH and this is beneficial for the reduction of hepatic failure after LH.

Objective To investigate the expression and prognostic significance of inhibitor of growth 4 (ING4) and tail-type homeobox transcription factor 2 (CDX2) in colorectal cancer. Methods The expressions of ING4 and CDX2 pro-teins were detected by immunohistochemistry in 99 tissue samples of colorectal cancer and 30 corresponding para-cancer-ous normal tissue samples. The data of clinic outcomes were collected. The correlations between the expressions of ING 4 and CDX2 and clinicopathological parameters were also analyzed. Results The positive expression rates of ING4 and CDX2 were 68.8%and 72.7%in colorectal cancer tissues, which were significantly lower than those of corresponding normal tissue samples (93.3% and 96.7%, P<0.05). There were significant differences in the differentiation, depth of invasion, lymph node metastasis and tumor stage between expressions of ING4 and CDX2 (P<0.05). The 5-year survival rate was significant-ly lower in ING4 negative group (35.5%) compared with that of ING4 positive group (77.9%). The 5-year survival rate was significantly lower in CDX2 negative group (48.1%) than that of CDX2 positive group (70.8%, P<0.05). The expression of ING4 was positively correlated with the expression of CDX2 in colorectal cancer. Conclusion The expressions of ING4 and CDX2 are strongly associated with the carcinogenesis, development and prognosis of the colorectal cancer,which suggests that ING4 and CDX2 might be used as prognostic markers for the colorectal cancer.

Objective To investigate the alteration of C/EBP α,C/EBP β and endoplasmic reticulum stress (ERS) related molecules (IRE1α and sXBP1) expression in pancreatic tissues in rats with hypertriglyceridemia related acute pancreatitis.Methods Ninety six Sprague Dawley (SD) rats were divided into 4 groups:control group,hypertriglyceridemia (HTG) group (n =24,fed with high fat diet for 2 weeks),AP group (n =24),HTG + AP group (n =24),and AP was induced by peritoneal injection of cerulein.The rats were sacrificed at 3,6,9,24 h after AP induction,respectively.The pathological changes of the pancreatic tissues were observed and scored by HE staining.Plasma levels of IL-1β,IL-6 and TNF-α were measured by ELISA.The expression of IRE1α,sXBP1,C/EBPoα,C/EBPβ mRNA were analyzed by real time PCR.The expressions of IRE1α,sXBP1,NF-kB,C/EBPα and C/EBPβ protein were determined by Western Blot.The expressions of C/EBPα and C/EBPβ proteins were also determined by immunohitochemistry.Results After two weeks of high fat diet,serum levels of triglyceride(TG) and total cholesterol (TC) in HTG group,HTG + AP group were much higher than those of the control group,and the difference was statistically significant (P ＜ 0.05).The pancreatic tissue injury was more severe in HTG + AP group,particularly at 9 h (P ＜ 0.05).And plasma IL-1β,IL-6 and TNF-α levels were also much higher in HTG + AP group when compared with that of AP group,the differences were all significant at 9 h (P=0.011;P=0.034;P =0.027).After AP induction,IRE1,sXBP1,C/EBP and C/EBPβ mRNA began to be up-regulated at 3 h,and IRE1 mRNA reached the highest level at 24 h,sXBP1 mRNA at 9 h,while C/EBP and C/EBPβ mRNA reached the highest level at 6 h.Compared with AP group,IRE1,sXBP1,C/EBP and C/EBPβ mRNA levels were much higher in HTG + AP group.In addition,as to IRE1 and sXBP1 mRNA,the difference was significant at 3,6,9,24 h,and C/EBP mRNA at 6,9,24 h,C/EBPβ mRNA at 6 and 9 h (P ＜ 0.05).After AP induction,IRE1α,sXBP1 and NF-kB proteins in the pancreatic tissue began to be up-regulated at 3 h,and all reached the highest level at 9 h.IRE1α,sXBP1 and NF-kB proteins were up-regulated more obviously in HTG + AP group,and the up-regulation in HTG + AP group was higher than that in AP group,and the high expressions of C/EBPα and C/EBPβ proteins could only be detected at 6 and 9 h in the HTG + AP group,while there was no expression detected in AP group.Conclusions C/EBPα,C/EBPβ,IRE1α and sXBP1 may be involved in the pathogenesis of HTG related AP,and IRE1α/sXBP1 pathway and C/EBPoα,C/EBPβ may mediate the pathologic injury and inflammation process of HTG related AP.

Objective To observe clinical response of the cladribine in the treatment of hairy cell leukemia.Methods Three patients were treated with cladribine 10 mg ivgtt for 3 or 5 days.Results Among 3 patients,2 patients achieved complete remission and 1 patient achieved near complete remission.Conclusion Cladribine has high efficacy and a favorable toxicity when adminisered to patients with hairy cell leukemia.

Objective: To evaluate the tolerance and safety of a human-mouse chimeric anti-CD20 monoclonal antibody IBI301 in Chinese patients achieved objective response with CD20⁺ B-cell non-Hodgkin’s lymphoma (NHL). Methods: Nine patients with CD20⁺ B-cell NHL received dose-escalating IBI301 infusions (250 mg/m², n=3; 375 mg/m², n=3; 500 mg/m², n=3, respectively). The data of all patients were collected for safety analyses. The median exposures of 125 mg/m², 375 mg/m², 500 mg/m² dose groups were 243, 690 and 980 mg, respectively. Safety and tolerability were evaluated by monitoring adverse events (AE). The ratios of CD19⁺, CD20⁺ B cells and the levels IgG and IgM were detected to evaluate the pharmacodynamics. Results: Totally 52 events of AE were observed, including 18 events of AE in 125 mg/m² group, 14 events of AE in 375 mg/m² group and 20 events of AE in 500 mg/m² group, respectively. There were 26 adverse reactions of 52 cases of AE, 22 reactions were judged to be probably related to IBI301, and 4 reactions were not probably related to IBI301, all disappeared or returned to baseline levels. Common AE in this study included decreased WBC, upper respiratory infection, decreased neutrophil count, dyspepsia, hyperuricemia, paresthesia, oral mucositis and dizziness. No patients quitted or trial discontinued. No severe AE (SAE) were reported. No dose-limiting toxicity (DLT) events were observed in the study. The ratio of CD20⁺ and CD19⁺ B cells decreased in all subjects. There was no significant changes of the levels of IgG and IgM. Conclusions: The single dose of IBI301 injection was well tolerated, and the AE occurred in the patients recovered. No SAE were reported, No DLT events were observed in the study. The IBI301 caused an elimination of the peripheral CD20-expressing B cells in all patients. Clinical trial registration Chinadrugtrials, CTR20140762.

Objective: To observe the material basis and mechanism of action of Kai-Xin-San (KXS) in regulating antidepression of neurotrophic factors. Methods: KXS eluted by ethanol on macroporous resin was prepared. The antidepressive effect of different components was compared by tailing suspension test and forced swimming test of mice. The levels of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) in hippocampus were determined by ELISA. The rat astrocyte glioma C6 cell line and the rat adrenal pheochromocytoma PC12 cell line were used to evaluate the effects of different ethanol elution sites on the expression of NGF and BDNF and the differentiation of PC12 cells.Results: All of the ethanol elution components from KXS exerted anti-depressive effects by shorting the immobile time of tailing suspension and forced swimming of mice and 70％ ethanol elution components exerted best efficacy. This site also could increase expressions of NGF and BDNF on C6 glioma cell line. The 10％ ethanol elution site had the strongest ability to promote PC12 cell differentiation. Ginsenosides were the main effectuve ingredients for promoting the expression of neurotrophic factors. Conclusion: Regulation of neurotrophic factors might be the prominent action mechanism of KXS exerting anti-depressive effects.

Cystic echinococcosis (CE) caused by E. granulosus is a serious helminthic zoonosis in humans, livestock and wildlife. Xinjiang is one of high endemic province for CE in China. A total of 55 sheep and cattle livers containing echinococcal cysts were collected from slaughterhouses in Changji and Yining City, northern region of Xinjiang. PCR was employed for cloning 2 gene fragments, 12S rRNA and CO1 for analysis of phylogenetic diversity of E. granulosus. The results showed that all the samples collected were identified as G1 genotype of E. granulosus. Interestingly, YL5 and CJ75 strains were the older branches compared to those strains from France, Argentina, Australia. CO1 gene fragment showed 20 new genotype haploids and 5 new genotype haplogroups (H1-H5) by the analysis of Network 5.0 software, and the YLY17 strain was identified as the most ancestral haplotype. The major haplotypes, such as CJ75 and YL5 strains, showed identical to the isolates from Middle East. The international and domestic trade of livestock might contribute to the dispersal of different haplotypes for E. granulosus evolution.
Abattoirs ; Animals ; Argentina ; Australia ; Cattle ; China ; Clone Cells ; Cloning, Organism ; Echinococcosis ; Echinococcus granulosus ; Echinococcus ; France ; Genetic Variation ; Genotype ; Haploidy ; Haplotypes ; Helminths ; Humans ; Liver ; Livestock ; Middle East ; Polymerase Chain Reaction ; Sheep