Objective To construct the recombinant expression vector carrying human glucocorticoid receptor ?(hGR?) cDNA to provide experiences for cloning of large DNA fragments and offer experimental materials for studying the function and medchanism of hGR? by antisense nucleic acid technique. Methods hGR? cDNA was amplified from an expressional phage library of human white cells by L-PCR, and cloned into pGEM-Teasy vector. Subsequently, the hGR? cDNA fragment digested from the recombinant T vector was sub-cloned into retroviral vector pLXSN in sense and antisense orientations, respectively through an improved ligasing protocol. Results Restriction endonuclease analysis and sequencing showed that cloning and sub-cloning of hGR? were successful. Conclusion Retroviral recombinant vectors carrying sense-and antisense-hGR? cDNA were successfully constructed.

Central South University began to implement the eight-year medical education in 2004, and has accumulated rich experience in the pre-medical education through continuous reform and improvement. Since 2012, Central South University has made a series of reformation, which is more conducive to the all-round development of medical students , on the pre-medical education . Through adding freshman courses, humanities courses, bilingual teaching courses, and applying aca-demic adviser institution, early scientific research training and open courses during the pre-medical education, Central South University has exercised and strengthened students' scientific research ability, the humanistic quality, English level, and made the pre-medical comprehensive examination for the shunt selection of students. Thus Central South University has improved the eight-year medical students' comprehensive quality, and provided an example of the reformation for medical education.

Objective To retrospectively explore the incidence and causes of apnea after operation for retinopathy of prematurity(ROP). Methods The clinical data of 17 premature infants with operation for ROP(ROP group)and the other 23 premature infants without ROP(control group)were collected.The occurrence of apnea(time of onset and remission and original diseases) was recorded after operation in ROP group and at adjusted 37 weeks of gestational age in control group.Follow-up was conducted for 4 weeks,and the prevalences of apnea were compared between these two groups.Results There was significant difference in prevalenees of apnea between ROP group and control group ( 52.9%vs 21.7%,P<0.05).In ROP group,time of occurrence of apnea was (38±40)h after operation,and time of remission was (126±145)h after onset.Among the infants with apnea,there were 3 cases of pneumonia(33.3%),1 case of hypoglycemia(11.1%)and 5 cases with unexplained causes(55.5%). Conclusion Infants after operation for ROP are more prone to apnea,and pediatricians and ophthalmologists are required to collaborate in the perioperative care.

[ ABSTRACT] AIM:To investigate the effect of the overexpression of voltage-gated chloride channel family protein 3 ( ClC-3) gene on bones of mice .METHODS: The tail gene detection assay was used to confirm the overexpression of ClC-3.The male FVB mice of three months old were divided into two groups , the wild type ( WT) group and the ClC-3 overexpressed (ClC-3 transgene) group.The body weight, length and weight of the right tibias were measured .The upper and middle parts of the tibias were dissected , decalcified, paraffin-imbed, sectioned and stained with HE staining .The bone morphology metrology was used to analyze the changes of bone structures .The percent trabecular area (%Tb.Ar), trabecular number ( Tb.N) , trabecular width ( Tb.Wi) and trabecular separation ( Tb.Sp) of cancellous bone in the upper part of the tibia were measured.The total tissue area (T.Ar), cortical area (Ct.Ar), percent cortical area (%Ct.Ar), marrow area ( Ma.Ar) and percent marrow area (%Ma.Ar) of the cortical bone in the middle part of the tibia were detec-ted .RESULTS:The wild type mice and the ClC-3-overexpressed mice were verified by the tail gene detection assay . Compared with WT group , the body weight and the length and weight of the tibia were decreased in ClC -3 transgene mice (P<0.05).In the cancellous bones of ClC-3 transgene mice, the%Tb.Ar and Tb.Wi were decreased (P<0.05), the Tb.Sp was increased (P<0.05) and the Tb.N was not significantly changed .In the cortical bones of ClC-3 transgene mice, the T.Ar, Ct.Ar and%Ct.Ar were decreased (P<0.05), the%Ma.Ar was increased (P<0.05), and the Ma. Ar was not significantly changed .CONCLUSION:ClC-3 overexpression may lead to the reduction of the bone mass and the destructure of the cancellous and cortical bones .The results suggest that ClC-3 may be involved in the regulation of bone resorption and/or formation.

Objective To summarize the clinical manifestations,diagnosis and treatment of perinatal tuberculosis.Methods We retrospectively analyzed the clinical data of six infants with perinatal tuberculosis treated in Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from January 2000 to December 2016.Results (1) The six patients were at the age of 27 to 60 days at the onset,while the mean ages of full-term and preterm infants were (31 ± 4) and (55± 6) days,respectively.Three mothers were diagnosed with active tuberculosis (two of which were diagnosed after parturition).(2) Three infants were admitted to the hospital with fever including two having respiratory diseases,one with external auditory canal secretions and deviated mouth,and two with secretions in the external auditory canal and masses in the head and neck.One was diagnosed as hepatosplenic tuberculosis,three as miliary tuberculosis (one was complicated by tuberculous meningitis and one with tuberculous meningitis and cervical lymph node tuberculosis),one with left cervical and temporal tuberculosis,and one with miliary tuberculosis complicated with left temporal,external auditory canal and cervical lymph node tuberculosis.(3) Two cases were positive for purified protein derivative skin test and two were positive for tuberculosis infection T cell spot test.Five cases were negative in anti-tuberculosis antibody test.Typical miliary shadows in the lungs of three infants were shown on chest X-ray.Two cases received auriculotemporal CT/MRI,indicating space-occupying lesions and destruction in the middle ear and mastoid process.Cranial MRI findings indicated one case had tuberculous meningitis.Diagnosis of three cases were confirmed by biopsy or after operation.(4) All six patients survived.Four cases recovered after combined treatment of isoniazid,rifampicin and pyrazinamide and two were transferred to a specialized hospital for further treatment.(5) All six patients were fully recovered in follow-ups at 2-3 years after discharge with normal development comparing with healthy children of the same age.Conclusions Perinatal tuberculosis,with nonspecific clinical manifestations,has a low incidence of infection but a high mortality rate.Suspected cases require full consideration including clinical symptoms,dynamic observation of laboratory findings and imaging features,and pathological examination when necessary.Maternal history of tuberculosis should be consulted and timely treatment is mandatory to improve the outcomes.

Objective:To assess the value of acoustic radiation force impulse (ARFI) elastography in the diagnosis of children with biliary atresia.Methods:A prospective survey of infants with hepatitis syndrome and hyperbi-lirubinemia in Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine from January 2017 to December 2018 was performed.The children with hepatitis syndrome were divided into the biliary atresia group( n=45) and non- biliary atresia group( n=30). Thirty children with hyperbilirubinemia were selected as the control group.Shear wave speed (SWS) of all infants was collected by ARFI ultrasound and compared among 3 groups.Receiver ope-rating characteristic curve(ROC curve) was used to analyze the optimal threshold value for SWS in the diagnosis of biliary atresia. Results:The mean SWS values in the biliary atresia group, non-biliary atresia group and the control group were (1.79±0.29) m/s, (1.26±0.12) m/s and (1.08±0.06) m/s, respectively.Compared with the control group, the mean SWS values in the biliary atresia group and non-biliary atresia group were significantly higher ( t=165.43, 15.75, all P<0.05). The mean SWS value in the non-biliary atresia group was significantly lower than that in the biliary atresia group ( t=90.27, P<0.05). With the non-biliary atresia group as reference, the area under the ROC curve of SWS for diagnosis of biliary atresia was 0.98(95% CI: 0.95-1.00), the optimal threshold was 1.45 m/s, and the sensitivity and specificity were 88.9% and 96.7%, respectively. Conclusions:Rapid non-invasive ARFI elastography is effective in the diagnosis of biliary atresia, and thus has important value for early diagnosis and treatment in clinical practice.

Objective To summarize the outcomes and clinical experiences of renal transplantation in human immunodeficiency virus (HIV)-positive patients .Methods The clinical data were retrospectively analyzed for one HIV-positive case of renal transplantation .Diagnosed as chronic renal insufficiency 1 year ago ,he received hemodialysis .After a positive screen for HIV ,he received highly active antiretroviral therapy (HAART) and HIV RNA turned negative 3 months later .CD4 + T cell count was 331 cell/μl at pre-operation and there was no HIV-rated opportunistic infection or cancer . Her mother donated her kidney . Basiliximab and steroid pulse therapy were used preoperatively and immunosuppressants were used after transplantation , including tacrolimus , corticosteroids and mycophenolate mofetil .Results The kidney was transplanted successfully and serum creatinine declined to a normal level at day 4 after transplantation .Because of an interaction between efaverenz and tacrolimus ,the blood concentration of tacrolimus was extremely low and the dose of tacrolimus had to be raised to 0 .2 mg/(kg·d) .Antiroviral therapy remained unchanged .No rejection and other complications were observed .And HIV RNA remained negative .Conclusions Renal transplantation is optimal for HIV-positive patients whose HIV status is completely under control .However ,drug interactions needs to be considered during perioperative and postoperative periods .

Children and adults differ significantly in physiology,biochemistry and immune function,which leads to sig-nificant differences in blood transfusion strategies between children and adults.To guide the clinical transfusion practice of pediatric patients and improve the prognosis of children,the National Health Commission organized the formulation and re-lease of the health industry standard Guideline for Pediatric Transfusion(WS/T 795-2022).This paper will briefly introduce some concepts that help understand of the Standard and the preparation process of the Standard,and explain and interpret the preparation of the"scope","general provisions"and"factors to consider"of the Standard,hoping to contribute to the understanding and implementation of the Standard.

OBJECTIVE: To predict the targets and pathways in the therapeutic mechanism of Guizhi Gancao Decoction (GZGCD) against heart failure (HF) based on network pharmacology. METHODS: The chemical components of GZGCD were analyzed using the databases including TCMSP, TCMID and TCM@Taiwan, and the potential targets of GZGCD were predicted using the SwissTargetPrediction database. The targets of HF were obtained using the databases including DisGeNET, Drugbank and TTD. The intersection targets of GZGCD and HF were identified using VENNY. Uniport database was used to convert the information, and the components-targets-disease network was constructed using Cytoscape software. The Bisogene plug-in, Merge plug-in, and CytoNCA plug-in in Cytoscape software were used for protein-protein interaction (PPI) analysis to acquire the core targets. Metascape database was used for GO and KEGG analysis. The results of network pharmacology analysis were verified with Western blot analysis. Three factors (PKCα, ERK1/2 and BCL2) were screened according to the degree value of network pharmacology results and the degree of correlation with heart failure process. The pentobarbtal sodium was dissolvein H9C2 cells treated with serum-free high glucose medium to simulate the ischemic anoxic environment of heart failure. The total proteins of myocardial cells were extracted. The protein contents of PKCα, ERK1/2 and BCL2 were determined. RESULTS: We identified a total of 190 intersection targets between GZGCD and HF using Venny database, involving mainly the circulatory system process, cellular response to nitrogen compounds, cation homeostasis, and regulation of the MAPK cascade. These potential targets were also involved in 38 pathways, including the regulatory pathways in cancer, calcium signal pathway, cGMP-PKG signal pathway, and cAMP signal pathway. Western blot analysis showed that in an in vitro H9C2 cell model of HF, treatment with GZGCD downregulated PKCα and ERK1/2 expressions and upregulated BCL2 expression. CONCLUSION The therapeutic mechanism of GZGCD for HF involves multiple targets including PRKCA, PRKCB, MAPK1, MAPK3, and MAPK8 and multiple pathways including the regulatory pathway in cancer and the calcium signaling pathway.
Humans ; Protein Kinase C-alpha ; Network Pharmacology ; Heart Failure/drug therapy* ; Proto-Oncogene Proteins c-bcl-2