Objective To construct humanized monoclonal antibodies against CD 20 and check their affinity to CD 20 antigen and their anti-tumor activity.Methods Based on the computer model , human IgG1 candidates closest to rituximab in crystal structure were selected in the Protein Data Bank ( PDB) .With the selected human IgG 1 candidates as the frame , we modified and transplanted the complementarity determining region ( CDR) of rituximab .First,the target gene fragments were obtained by overlapping PCR.Then, the sequences of the light chains(L) and the heavy chains(H) were inserted in-to the pcDNA3.3 and pOptiVEC vectors.Next, the constructed clones were transfected into 293F cells through transient transfection.After a large-scale cell culture, the mAb was purified by affinity chromatography rProtein A column.The puri-ty and expression level of the humanized antibodies was tested by sodium dodecyl sulfate ( SDS)-polyacrylamide gelelectro-phoresis(PAGE).The affinity of the humanized antibodies to CD20 was assessed with Fortebio assay.Finally, the anti-tumor activity of the constructed antibodies was detected by checking the tumor growth inhibition of the nude mice transplan-ted with tumor .Results Three humanized monoclonal antibodies against CD 20 were expressed and purified successfully . In reducing SDS-PAGE, the antibodies exhibited two bands of approximately 25 ×103 and 55 ×103 , respectively.The band size of the antibodies matched the expected value.Fortebio assay revealed that the humanized antibodies could bind to CD20 with high affinity (rituximab:6.48 ×10 -9mol/L, L4H7:1.91 ×10 -9mol/L, L5H5:7.35 ×10 -10mol/L,and L5H7:1.91 ×10 -9mol/L).The tumor growth inhibition experiment showed that the anti-tumor activity of L5H7 mAb was better than that of rituximab .Conclusion Three humanized monoclonal antibodies against CD 20 have been successfully construc-ted and expressed.L5H7 mAb possesses high affinity for CD20 and a good ability to kill tumor cells.

Objective To investigate the leukemia incidence among residents in the vicinity area of a nuclear installation.Methods The data were collected by the retrospective survey.Diagnosis is based on the medical records in all the relevant hospitals.The study area was those residential areas within 10 km of radium surrounding the installation.All residents who had lived in the study area for six months and over were recruited into the study group.Results The survey ascertained 643 newly diagnosed leukemia cases in the study area during 1997 to 2007.The crude rate was 2.51/100,000,and its standardized rate to national population structure was 2.53/100,000.The incidence rate in the study area was slightly lower than that in the whole city.No increase trend was detected during the period.Conclusions No significant difference of the leukemia incidence rate was found between the vicinity area of the nuclear installation and the whole city.

Objective To evaluate the efficacy and safety of transarterial chemoembolization (TACE)combined with sonographically guided percutaneous microwave coagulation therapy(PMCT)for hepatic carcinoma with diameter＞5.0 cm.Methods We retrospectively reviewed 68 cases of hepatic carcinoma with diameter＞5.0 cm under treatment of TACE combined with PMCT.CT,USG and correlated laboratory tests of hepatic carcinoma were carried out.Results Among 68 cases,complete ablation were 5 cases(5/68),tumor ablation area more than 50% or tumor shrinkage less than 30% were 59 cases(59/ 68),tumor ablation area less than 50% or tumor shrinkage more than 30% were 6 cases(6/68).Forty five cases with high AFP descended more than 50% after the procedure in 42 eases(93.33%).Thirty seven cases and 29 cases with increase of CEA and CA19-9 decreased to 28(75.97%)and 23(93.10%)cases with corresponding index decreasing more than 50% respectively.Survival time reached 4-6 months in 3 cases, more than 6 months for 31 cases,more than 12 months of 34 cases.Two cases among them showed no recurrence up to now after stoppage of treatment for 24 months and finally no correlative mortality occurred. Conclusion TACE combined with sonographically guided PMCT for hepatic carcinoma with diameter more than 5 cm is safe and effective.

Benzene ; adverse effects ; analysis ; chemistry ; Catalysis ; Humans ; Occupational Exposure ; Risk Assessment ; Vehicle Emissions ; analysis ; Workplace

Parkinson's disease (PD) is the second major neurodegenerative disease.The pathogenesis of PI) is still unclear.It is generally believed that neural damage, mitochondrial dysfunction, inflammation, oxidative stress and autophagy dysfunction caused by the transmission and aggregation of a- synuclein play an important role in the occurrence and development of PD.More and more research show- that metabolic disorder is one of the pathogenesis of PD.We examined whether overexpression of a- synuclein could induce metabolic disorder in mice and the possible mechanisms.Mice were divided into two groups: Thyl-aSYN transgenic mice (TG) and the control wild-type (WT) group.The rotarod test was used to analyze motor function in mice.We detected the body weight, plasma insulin content, glucose tolerance and insulin tolerance in the two group mice.The morphology of islets in the two groups were observed by hematoxylin eosin (HE) staining, and the islets were isolated to detect the glucose- stimulated insulin secretion (GSIS).The results showed that compared with the WT group, exercise tolerance of 12-month-old TG group decreased by 23.1% (P < 0.05) , body weight increased by 7% (P < 0.01), glucose tolerance decreased (P < 0.05), insulin tolerance decreased (P < 0.05), and insulin contents in the peripheral blood decreased by 20% (P < 0.05).Compared with the WT group, the levels of ce -syn proteins in the pancreas of the TG group increased by 1.32 times (P < 0.05) , the area of islets in the TG group decreased (P < 0.05 ) , the number of islets decreased (P < 0.01) , and the insulin secretion function decreased (P< 0.01).This study showed that the role of a-synuclein in PD is not limited to the damage of dopaminergic neurons, it also can affect metabolism and the morphology and function of peripheral organs, which provides a new theoretical basis for the pathogenesis of PD.

Previous studies have demonstrated that plasminogen actiator (PA) activity is significantly high in semen of infertile men, which is also high in semen when azoospermia or oligozoospermia is induced by injection of testosterone enanthate (TE) into healthy adult men or rhesus monkeys. To further clarify the source and possible role of PA in semen, the present study was undertaken to examine: (1) whether the mRNAs for tissue type PA (tPA), urokinase type PA (uPA), and PA inhibitor-1 (PAI-1) are expressed in epididymis, seminal vesicle and prostate gland of rhesus monkeys; and (2) whether PA has some effect on in vitro sperm capacitation as judged by the potential of sperm motility, acrosome reaction (AR) and in vitro fertilization. Our results showed that (1) mRNAs for PA and PAI-1 were expressed in epididymis, seminal vesicle and prostate gland, and (2) uPA, but not tPA, improved sperm mobility, induced AR and enhanced sperm capacity to fertilize mature eggs. Thus, it is concluded that PA activity in semen comes not only from testis and epididymis, but also from seminal vesicle and prostate gland; and that uPA, but not tPA, may play a role in sperm capacitation.

Objective To compare the spiral CT and pathological findings of sub-centimetre pulmonary ground glass nodules(GGNs), and to improve the aaccuracy in early diagnosis.Methods The CT findings and pathologic features of 30 patients with sub-centimetre pulmo-nary GGNs in our hospital from May 2013 to June 2016 were reviewed retrospectively.The patients were divided into 3 groups according to their pathological classification,namely the benign group,the preinvasive group and the invasive group.The shape,size,margin,vacuole sign, pleural indentation,the relations with bronchi and blood vessels of the pulmonary GGNs were analyzed.Results There was no significant difference in shape,maximum diameter and margin between the benign group and the other two groups(P>0.05).Adjacent vessels changes of the lesion showed significant differences between benign group and preinvasive or invasive groups(P<0.05).And there were significant differences in size,pleural indentation,vacuole sign and adjacent vessels changes between the preinvasive group and the invasive group(P<0.05).Conclusion The comprehensive analysis of lesions size,vacuole sign,pleural indentation and adjacent vessels changes can be helpful to improve the aaccuracy of differential diagnosis on sub-centimetre pulmonary ground glass nodules.

Objective To observe the efficacy and safety of applying sirolimus-eluting stents in emergency percutaneous coronary intervention (PCI) for the patients with acute myocardial infarction (AMI).Methods In total,220 patients with AMI were enrolled in this study at Shanghai Tongji Hospital, divided into two groups,one with bare-metal stent and the other with sirolimus-eluting stent.Cardiovascular fatality,major adverse cardiac events (MACE) and target vessel revascularization (TVR) were observed one and six months after PCI in the two groups.Results There was no significant difference in overall fatality and MACE in the 1~(st) or 6~(th) months after PCI between the two groups.Three cardiogenic deaths occurred in bare-metal stent group with a fatality of 2.8 percent,and five deaths in sirolimus-eluting stent group with a fatality of 4.5 percent in six months after PCI.However,rate of restenosis in those with sirolimus-eluting stents was significantly lower than that of bare-metal stents (6.0 percent vs 16.1 percent,P

Adolescent ; Adolescent Development ; Child ; Child Development ; China ; epidemiology ; Female ; Humans ; Male ; Obesity ; epidemiology ; Prevalence

OBJECTIVETo observe the effect of chronic lead contaminant on mRNA expression of protein kinase C (PKC) and calmodulin (CaM) in hippocampus of baby rats.

METHODSThe Wistar pregnant rats were randomly divided into 3 groups fed with distilled water and lead contained water (0.2% and 1.0% lead acetate) respectively. The lead exposure period was from the 0 day of pregnancy to the day when the offspring weaned. Then the baby rats were fed with lead water the same as their mothers. The cliff avoidance reflex within postnatal day 8 and step down test at postnatal day 50 were performed. Then pups were killed at postnatal day 8 and 50 respectively. Atomic absorption spectrometry was used to determine lead content of rats' brain. RT-PCR was used to observe mRNA expression of PKC and CaM in hippocampus of baby rats.

RESULTSThe brain lead content of test groups were much higher than that of the control group. The completion rate of cliff avoidance reflex and the score of step down test of test groups were lower than those in the control group (P < 0.05). Compared with control group, PKC and CaM mRNA expression of chronic lead exposure baby rats in the hippocampus had the down trend (P < 0.05).

CONCLUSIONThe decrease of PKC and CaM mRNA expression level in hippocampus has a great link with the impairment of learning and memory induced by lead in baby rats, which might be one of the molecule mechanisms of lead induced impairment of learning and memory.

Animals ; Calmodulin ; genetics ; metabolism ; Female ; Hippocampus ; drug effects ; metabolism ; Lead ; toxicity ; Learning ; drug effects ; Male ; Memory ; drug effects ; Protein Kinase C ; genetics ; metabolism ; RNA, Messenger ; genetics ; Rats