Objective:To investigate the potential value and feasibility of MG132 as a new therapy method for the ovarian cancer and a cisplatin chemotherapy-synergistic agent.Methods:The mode of the nude mice transplanted tumor tissues of ovarian epithelial carcinoma were established,20 nude mice were randomly divided into four groups and were all intrapedtoneal injected,once a day,a total of seven days:①Control group(0.2 ml saline),②MG132 group(2 mg/kg),③Cisplatin group(1 mg/kg),④Combination group[MG132(2 mg/kg) + cisplatin (1 mg/kg)].The weight inhibitory rates of tumors in each group were compared after four weeks.The expressions of Caspase3,Beclin1 in each group were detected by IHC,FIA,western blot and RT-PCR.Results:①The inhibitory rate of tumors in cisplatin group,MG132 group,and combination group was 53.85％,15.38％,88.46％,respectively,the additive effect of cisplatin and MG132 combination therapy was 60.95％.②IHC,FIA,western blot detected that compared to control group,the positive expressions of Beclin1,Caspase3 were increased in cisplatin group,MG132 group,and combination group,among which combination group increased more.③RT-PCR detected that the mRNA relative quantity of Beclin1 in cisplatin group,MG132 group,combination group respectively were higher than that of control group(P＜0.05);and it was higher in combination group than that of cisplatin group and MG132 group(P＜0.05).Conclusions:The growth of nude mice transplanted tumor tissues of ovarian epithelial carcinoma can be inhibited by MG132,and has a synergistic effect for treating ovarian cancer by combination with cisplatin,it is expected to be an effective anti-tumor drug for platinum resistant refractory ovarian cancer.

A survey was conducted to investigate the situation of English works reading activities among medical students and their attitude to the medical contents in English works.The result showed that medical students support the setting-up of literature and medicine course and it was believed that medicine in literature would be helpful to improve the quality of medical humanity education.

Objective To evaluate the value of diffusion.weighted imaging(DWI)in the differential diagnosis of benign and malignant breast lesions with Meta.analysis.Methods English and Chinese articles published from 1997 to 2007 were searched in Cochrane library,Pubmed,Ovid.Elsevier.Springer databases and Chinese National Knowledge Infrastructure(CNKI),and selected according to the criteria for diagnostic research recommended by Cochrane Method Group.The characteristics of selected articles including the study background,design,results and diagnostic indicators were analyzed.Statistical analysis was performed with Meta-test version 0.9.Results Six articles were included with a total of 479 lesions.The sensitivity and specificity of DWI for breast 1esions ranged from 64.0% to 92.8% and from 45.8% to 96.7% respectively. All groups had homogeneity.and a fixation effects regression Was developed.The pooled weighted sensitivity and specificity with 95% CI and Az Was 86%(76%-92%).80%(61%-91%)and 92.19% respectively.Sensitivity analysis demonstrated no remarkable change on heterogeneity.Condnsion The value of ADC in DWI can help to differentiate the benign breast lesions from the malignant with good sensitivity and specificity.The further study is needed to assess the clinical aplication of the b-value and diagnostic threshold.

OBJECTIVETo investigate whether DTI abnormalities in patients with nondemented subcortical ischemic vascular disease (SIVD) are correlated to general cognitive function and executive function independent of conventional MRI parameters.

METHODSSixty-six SIVD patients including 30 with cognitive impairment without dementia (VCIND) and 36 with normal cognition (NCI) underwent DTI and cognitive assessment of the general cognitive function and executive function. Conventional MRI parameters and DTI parameters were measured within the white matter lesions (WMLs), normal-appearing white matter (NAWM) and normal-appearing gray matter (NAGM). The independent predictors of general cognitive function and executive function in SIVD patients was analyzed.

RESULTSNCI and VCIND patients showed significant differences in the periventricular WMLs, ADC valus in NAWM and WMLs of the centrum semiovale, and FA values in NAWM of the anterior periventricular. Except for ADC values of the caudate nucleus, ADC and FA values in the subcortical NAGM showed significant difference between the two groups. After controlling for age and education, PVLs and ADC values in the WMLs and NAWM of the centrum semiovale and putamen, and FA values in the anterior periventricular NAWM, WMLs and putamen showed significant correlations to MMSE; the number of lacunar infarcts, ADC values in posterior periventricular NAWM and caudate nucleus, and FA values in anterior periventricular NAWM and thalamus showed significant correlations to CDT. Multivariate analysis showed independent correlations of the ADC values in WMLs and NAWM of the centrum semiovale to MMSE, and demonstarted correlations of the ADC values of the caudate nucleus and number of lacunar infarcts to CDT.

CONCLUSIONIn nondemented SIVD subjects, the integrity of the white matter in the centrum semiovale strongly correlates to the general cognition, and the microstructural damage of the caudate nucleus head predicts executive function impairment independent of other MRI variables.

Brain Ischemia ; pathology ; physiopathology ; psychology ; Case-Control Studies ; Caudate Nucleus ; pathology ; Cerebral Cortex ; pathology ; Cognition ; physiology ; Cognition Disorders ; pathology ; physiopathology ; Diffusion Tensor Imaging ; Humans ; Magnetic Resonance Imaging

Objective To observe the effect of immersion virtual reality(IVR)training com-bined with occupational therapy(OT)in the treatment of stroke patients with unilateral neglect.Methods Fifty stroke patients with unilateral neglect in Shanghai Fourth Rehabilitation Hospital were randomly divided into IVR plus OT group and OT group,with 25 cases in each group.The OT group received conventional OT for unilateral neglect,and the IVR plus OT group received IVR train-ing and OT.Both groups were treated for 4 weeks.Before and after treatment,the Catherine Bergego Scale(CBS),line cancellation test,star cancellation test,and drawing clock test were used to evalu-ate unilateral neglect symptoms;the Fugl-Meyer Assessment for Upper Extremity(FMA-UE)and Barthel index(BI)were used to evaluate motor function of upper extremity and activities of daily liv-ing.Results After treatment,the results of CBS,line cancellation test,star cancellation test,draw-ing clock test,FMA-UE and BI scores were significantly improved when compared with those before treatment in both groups,and the improvement effects in the IVR plus OT group were significantly better than those in the OT group(P＜0.05 or P＜0.01).Conclusion IVR training combined with OT can improve unilateral neglect symptoms,motor function of affected upper extremity,and activi-ties of daily living in stroke patients.

Objective To observe the effect of immersion virtual reality(IVR)training com-bined with occupational therapy(OT)in the treatment of stroke patients with unilateral neglect.Methods Fifty stroke patients with unilateral neglect in Shanghai Fourth Rehabilitation Hospital were randomly divided into IVR plus OT group and OT group,with 25 cases in each group.The OT group received conventional OT for unilateral neglect,and the IVR plus OT group received IVR train-ing and OT.Both groups were treated for 4 weeks.Before and after treatment,the Catherine Bergego Scale(CBS),line cancellation test,star cancellation test,and drawing clock test were used to evalu-ate unilateral neglect symptoms;the Fugl-Meyer Assessment for Upper Extremity(FMA-UE)and Barthel index(BI)were used to evaluate motor function of upper extremity and activities of daily liv-ing.Results After treatment,the results of CBS,line cancellation test,star cancellation test,draw-ing clock test,FMA-UE and BI scores were significantly improved when compared with those before treatment in both groups,and the improvement effects in the IVR plus OT group were significantly better than those in the OT group(P＜0.05 or P＜0.01).Conclusion IVR training combined with OT can improve unilateral neglect symptoms,motor function of affected upper extremity,and activi-ties of daily living in stroke patients.

To quantify the transcriptional activity of NF-κB and to screen drugs related to the regulation of NF-κB activation, we constructed a recombinant plasmid through deleting the original CMV promoter of retrovirus vector pQCXIP and inserting the NF-κB enhancer and NanoLuc luciferase sequence into the vector. Then, using the recombinant plasmid we constructed a cell line in which the expression of NanoLuc luciferase (NLuc) was regulated by NF-κB. The inserted sequences were verified by restriction endonuclease digestion and sequencing. Tumor necrosis factor-α (TNF-α), an NF-κB activator, acted on the constructed NLuc cell line and leaded to the specific luciferase reaction. The luciferase reaction showed a fine time and dose dependence to the TNF-α stimulation, indicating the successful construction of the NF-κB regulated NLuc-expressing cell line. Besides, the NF-κB inhibitor, triptolide, reduced the expression of NLuc in a dose-dependent way. The constructed reporter system in this study could be applied in the quantification of the NF-κB transcriptional activity and in the NF-κB regulation-related drug screening.

Objective The aim is to utilize CRISPR/Cas9 gene editing technology to construct Dmd gene mutant mice with a point mutation in exon 23 of the Dmd gene. Subsequently, the phenotypic changes of the mice in muscles and immune systems are analyzed and verified, providing an evaluation model for Duchenne muscular dystrophy and other related diseases.MethodsBased on the sequence characteristics of exon 23 of the Dmd gene, small guide RNA (sgRNA) was designed and synthesized. Cas9 mRNA, sgRNA fragments, and oligo donor DNA were microinjected into fertilized eggs of C57BL/6J mice. After transferring the fertilized eggs to surrogate mice, F0 generation mice were born. After mating with F0 generation mice, offspring mice were obtained, and Dmd gene positive mutant (Dmd^Mu/+) mice were obtained after genotype identification. Male hemizygous Dmd^Mu/+(Dmd^Mu/Y) mice were selected for phenotype validation. The body weight of live 3- and 9-month-old mice were recorded. Muscle tension was evaluated through the grid test. Hearts and semitendinosus muscles were collected, and the histopathological changes were observed using HE staining. Further, the expression of Dmd protein in muscle tissue of 9-month-old mice was analyzed by Western blotting.An acute inflammation model was established in Dmd^Mu/Y mice using lipopolysaccharide induction. Peripheral blood from the submandibular vein was collected, and the changes in the proportion of neutrophils and monocytes were detected by flow cytometry.Results The results of genome sequencing and Western blotting confirmed the successful construction of Dmd gene point mutant mice (Dmd^Mu/+ mice). Dmd protein expression was not detected in skeletal muscle and myocardium of Dmd^Mu/+ mice, and it was significantly reduced compared to wild-type C57BL/6J mice (P<0.05). Compared with wild-type mice of the same background, Dmd^Mu/Y mice at 3 and 9 months of age showed significant weight loss (P<0.01) and decreased muscle tension (P<0.05). 9-month-old Dmd^Mu/Y mice exhibited significant pathological changes in skeletal muscle and myocardium, including widening of intermuscular space. Under normal condition, compared with wild-type mice, the proportion of neutrophils and monocytes in the peripheral blood of 3-month-old Dmd^Mu/Y mice was significantly lower than that of wild-type mice (P<0.01). After lipopolysaccharide stimulation, the proportion of neutrophils in peripheral blood of 3-month-old Dmd^Mu/Y mice remained significantly lower compared to that of wild-type mice (P<0.01). The proportion of neutrophils in peripheral blood of 9-month-old Dmd^Mu/Y mice significantly decreased after lipopolysaccharide induction (P<0.01), with a trend of change observed in monocytes between groups.Conclusion The successful construction of the Dmd gene mutant mouse model has confirmed the vital function of Dmd gene in maintaining normal muscle tissue morphology and muscle tone. It preliminarily indicated that Dmd gene deletion could significantly reduce the proportion of neutrophils in peripheral blood, offering a new perspective for the study of immune system alterations in Duchenne muscular dystrophy patients.

Objective To monitor the antifungal resistance of clinical isolates of Candida spp.in the East China region.Methods MALDI-TOF MS or molecular methods were used to re-identify the strains collected from January 2018 to December 2022.Antifungal susceptibility testing was performed using the broth microdilution method.The susceptibility test results were interpreted according to the breakpoints of 2022 Clinical and Laboratory Standards Institute(CLSI)documents M27 M44s-Ed3 and M57s-Ed4.Results A total of 3 026 strains of Candida were collected,65.33％of which were isolated from sterile body sites,mainly from blood(38.86％)and pleural effusion/ascites(10.21％).The predominant species of Candida were Candida albicans(44.51％),followed by Candida parapsilosis complex(19.46％),Candida tropicalis(13.98％),Candida glabrata(10.34％),and other Candida species(0.79％).Candida albicans showed overall high susceptibility rates to the 10 antifungal drugs tested(the lowest rate being 93.62％).Only 2.97％of the strains showed dose-dependent susceptibility(SDD)to fluconazole.Candida parapsilosis complex had a SDD rate of 2.61％and a resistance rate of 9.42％to fluconazole,and susceptibility rates above 90％to other drugs.Candida glabrata had a SDD rate of 92.01％and a resistance rate of 7.99％to fluconazole,resistance rates of 32.27％and 48.24％to posaconazole and voriconazole non-wild-type strains(NWT),respectively,and susceptibility rates above 90％to other drugs.Candida tropicalis had resistance rates of 29.55％and 26.24％to fluconazole and voriconazole,respectively,resistance rates of 76.60％and 21.99％to posaconazole and echinocandins non-wild-type strains(NWT),and a resistance rate of 2.36％to echinocandins.Conclusions The prevalence and species distribution of Candida spp.in the East China region are consistent with previous domestic and international reports.Candida glabrata exhibits certain degree of resistance to fluconazole,while Candida tropicalis demonstrates higher resistance to triazole drugs.Additionally,echinocandins resistance has emerged in Candida albicans,Candida glabrata,Candida tropicalis,and Candida parapsilosis.

RANKL/RANK/OPG axis is important in bone metabolism regulation, and becomes a popular research area in bone diseases. RANKL is a critical part of RANKL/RANK/OPG axis, and widely required in bone metabolism research. However, the yield of recombinant soluble human RANKL (hRANKL) in Escherichia coli is much lower than mouse RANKL (mRANKL). In this study, by adjusting and stabilizing the pH value of LB medium at 7.5, lowering the inducing temperature to 16 ℃ and optimizing the lysis program, the yield of soluble hRANKL increased by approximately 5 to 12-fold over the non-adjusted group. Our experiment effectively enhanced soluble hRANKL expression in E. coli and might constitute a meaningful attempt to obtain soluble expression of recombinant protein in E. coli.