Background Heat shock proteins (HSPs) are highly conserved proteins that are induced in cells when confronted with a wide variety of proteotoxic stresses.HSP27 has a high degree of similarity with α-crystallin protein.The abnormality of HSP27 structure and expression are closely related to the formation of cataracts.Our previous study showed sodium salieylate has the protective effect on H2O2-induced lens damage.Objective This study was to investigate the roles of MAPK signal pathway in sodium salicylate-induced the expression of HSP27 in human lens epithelial cells (LECs) in vitro.Methods Human LECs were incubated in the fresh media containing sodium salicylate at different concentrations (0-55 mmol/L) for different times (1-5 hours) and allowed to be recovered in fresh medium without sodium salicylate for 1-24 hours with or without pretreatment with P38MAPK inhibitor (SB203580), ERK1/2 inhibitor (PD98059) and JNK/SAPK inhibitor ( SP600125). The expressions of P38MAPK, EBK1/2, JNK/SAPK, phosphorylated P38MAPK, phosphorylated ERK1/2, phosphorylated JNK/SAPK and HSP27 were detected by Western blot. HSP27 mRNA was detected by RT-PCR. The expression of HSP27 was also detected by immunohistochemistry. Results There was only weak expression of HSP27 in normal human LECs.After stimulation of 35-55 mmol/L sodium salicylate was removed and human LECs were cultured again for 6 hours,the expression of HSP27 in LECs were significantly increased ( F= 509. 953,P<0. 01). HSP27 was absent expressed in human LECs in 55 mmol/L sodium salicylate stimulation for 1-5 hours groups, but LECs were re-cultured for 3,6 hours after removed the stimulation, the expression of HSP27 was elevated (F = 452. 534, P<0. 01). Activation of P38 M APK occurred after sodium salicylate stimulation 30 minutes and 1 hour ( F = 865.68, P<0. 01). However, ERK 1/2 was expressed after sodium salicylate was eliminated for 1-6 hours ( F = 388.84, P<0. 01). JNK/SAPK was inactived by sodium salicylate. The expression of HSP27 could be down-regulated with the pretreatment of SB203580 and PD98059. Conclusion Sodium salicylatc can induce the expression of HSP27 in human (LECs) . The effects are mediated,at least in part ,through the activation of P38MAPK and ERK1/2 signaling pathway .

Objective To explore the clinical diagnosis and management of hepatecellular adenoma(HA). Methods Seventeen HA cases treated in our hospital from Janurary 1986 to June 2007 were studied retrospectively.The clinical and pathologic data were reviewed. Results Of 17 patients,9 were male and 8 were female with median age of 46 years.None of the female patients had a history of oral contraceptives.Most patients were agymptomatic.Preoperative liver function test and AFP serum test were normal.Preoperative imaging study yielded deftnite diagnosis in only a fraction of patients.All 17 cases underwent surgical resection and the postoperative course is uneventful.All cases were followed up ranging from 6 to 252 months.Recurrence was not found.Pathology reported atypical hyperplagia in 2 cases and with malignant transformation in 1 case. Conclusions The diagnosis of HA is exclusive and dependent on clinical data,laboratory test and radiography.HA has a tendency to malignancy.Surgery is recommended in all cases.

Objective To sum up the therapeutic results of 125 cases of tetralogy of Fallot(TOF),and ex- plore the optimal time and risk factors of opration,as well as perioperative management.Methods One hundred and thirth-one consecutive cases of TOF underwent corrective surgery.There were simple stenosis of infundibular portion in right ventricular outflow tract in 37 cases,stenosis of infundibulum and pulmonary valve in 14 cases,main pul- monary trunk and left/right pulmonary arteries stenosis in 74 cases,and pulmonary atresia in 5 cases.Autologousper- icardial conduit,or with waived were used for right ventricular outflow tract and right ventriculo-pulmonary artery connection.Other anomalies were corrected.Results The surgicalmortality was 4.0 %.The cause of death were se- rious low cardiac output syndrome(3 patients),respiratory function failure(1 patient),pericadial infection(1 pa- tient).Conclusion It is necessary to profonn corrective opration on younger TOF patients.Effetive prophylaxis and control of low cardiac output syndrome and pulmonary complication is a useful strategy.

In order to enhance the antitumor efficacy of recombinant Newcastle disease virus, rNDV-IL15 was rescued in this study. Recombinant plasmid prNDV-IL15 was constructed, and BHK21 cells were transfected with the recombinant plasmid. Finally, the recombinant Newcastle disease virus rNDV-IL15 was successfully rescued. The growth curves of these two recombinant viruses were determined. Murine melanoma B16F10 cells were infected with rNDV-IL15 at MOI of 0.1, and the expression level of IL15 in the supernatant was detected by ELISA. The antitumor efficacy of rNDV-IL15 and rNDV was compared in vitro and in vivo. Results showed that prNDV-IL15 was constructed and recombinant virus rNDV-IL15 was successfully rescued. The growth curve of rNDV-IL15 showed that the growth of rNDV-IL15 had not been changed after insertion of IL15 gene. Results showed that there was high level of IL15 expression in the supernatant of rNDV-IL5-infected B16F10 cells (1 044.3 +/- 27.7 ng x mL(-1)). rNDV-IL15 and rNDV significantly inhibited the growth of B16F10 cells in vitro in a time-dependent manner. However, there was no significant difference between them. In animal experiments, rNDV-IL15 efficiently suppressed tumor growth in vivo when compared with rNDV, and the difference was statistically significant. The results suggested that rNDV-IL15 is a more effective antitumor agent.
Animals ; Body Weight ; Cell Line, Tumor ; Cell Proliferation ; Chick Embryo ; Cytotoxicity, Immunologic ; Female ; Genetic Therapy ; Interleukin-15 ; genetics ; metabolism ; Melanoma, Experimental ; pathology ; therapy ; Mice ; Neoplasm Transplantation ; Newcastle disease virus ; genetics ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Tumor Burden

This study is to develop a therapeutic drug monitoring (TDM) network server of tacrolimus for Chinese renal transplant patients, which can facilitate doctor to manage patients' information and provide three levels of predictions. Database management system MySQL was employed to build and manage the database of patients and doctors' information, and hypertext mark-up language (HTML) and Java server pages (JSP) technology were employed to construct network server for database management. Based on the population pharmacokinetic model of tacrolimus for Chinese renal transplant patients, above program languages were used to construct the population prediction and subpopulation prediction modules. Based on Bayesian principle and maximization of the posterior probability function, an objective function was established, and minimized by an optimization algorithm to estimate patient's individual pharmacokinetic parameters. It is proved that the network server has the basic functions for database management and three levels of prediction to aid doctor to optimize the regimen of tacrolimus for Chinese renal transplant patients.
Algorithms ; Bayes Theorem ; Database Management Systems ; Drug Monitoring ; methods ; Humans ; Immunosuppressive Agents ; administration & dosage ; pharmacokinetics ; Kidney Transplantation ; Models, Biological ; Tacrolimus ; administration & dosage ; pharmacokinetics

OBJECTIVETo develop a rapid test kit for antibody to HIV by nano immunomagnetic lateral flow method.

METHODSA rapid test kit was developed by conjugation of the HIV antigen gp41 and gp36 to 200nm super paramagnetic particles by carbodiimide (EDC) and coating of the HIV antigen gp41 and gp36 to nitrocellulose membrane. Then the kit was evaluated with serials of experiments.

RESULTSThe kit was qualified with examination of national reference panel of anti-HIV antibody for colloidal gold diagnostic kit. The sensitivity was 100% by tested with 20 HIV antibody positive sera, the specificity was 98.5% by tested with 600 HIV antibody negative sera, respectively. The stability of the kit was over 12 month by storage at room temperature.

CONCLUSIONA diagnostic kit for antibody to HIV was developed with the advantages of convenience, rapid test, good stability and point of care.

Antibodies, Anti-Idiotypic ; immunology ; Gold Colloid ; chemistry ; HIV ; immunology ; isolation & purification ; HIV Antibodies ; HIV Envelope Protein gp41 ; HIV Infections ; diagnosis ; HIV Seropositivity ; blood ; HIV-1 ; immunology ; isolation & purification ; Immunomagnetic Separation ; methods ; Molecular Biology ; methods ; Nanotechnology ; Reagent Kits, Diagnostic

Background Leber hereditary optic neuropathy (LHON)is a mitochondrial DNA (mtDNA)hereditary disease,so it is significant to understand the influence of DNA mutation on the occurrence of LHON.Objective This survey was to evaluate the role of mtDNA mutation in the development of LHON.Methods This survey study was approved by the Ethic Committee of Wuhan General Hospital of Guangzhou Military Command and written informed consent was obtained from each subject before the relative medial examination.Seventy-two matrilineal relatives from a family with LHON were collected for a pedigree analysis and mutation screening.Regular eye examination was performed on 11 patients,13 mutant gene carriers and 49 individuals with normal phenotype,and the degree of visual damage was graded as follows: ＞0.3 was normal,0.1-0.3 was mild damage,＜0.05-0.1 was moderate damage,＜0.02-0.05 was severe damage and ＜0.01 was very severe damage.Clinical characteristics of LHON was evaluated.The periphery blood sample of 2-4 ml was collected from individuals to separate the mononuclear cells,and the mtDNA was extracted by modified high salt method.MtDNA was amplified by PCR and the mutation loci was sequenced.Results PCR amplification product sequencing of mutant gene showed that both G11778A and T14502C mutations were detected in 24 of 72 matrilineal relatives,but only 11 of 24 carriers developed LHON.No abnormal clinical findings were seen in the 13 carriers,showing a less 50％ penetrance in this family.There was no G11778A or/and T14502C mutation in the normal phenotype individuals of this family.The onset age for vision impairment in 11 affected matrilineal relatives varied from 8 to 50 years old,with the mean age of 24.36 years old,showing a significantly lower age than that of the 13 carriers (5-72 years old,mean 40.38 years old) (t =2.102,P=0.049).Conclusions This study suggests that the Gl1778A and T14502C mutation in mitochondrial DNA is one of causes in the development of LHON.The primary G11778A mutation together with T14502C mutation in mtDNA is a factor for the occurrence of LHON,hut it is not sufficient to the development of LHON.An effective “second hit” process will play an inducing role for LHON.

To investigate the phenotypic characteristics of the strain of the rabies virus CTNCEC25, the strain of the China rabies virus CTN-1 adapted to primary chicken embryo cells (CECs), Vero cells, and mouse neuroblastoma N2a cells was inoculated with CTNCEC25 and parental CTN-1 strains to explore the cytopathic effect (CPE) and growth kinetics of CTNCEC25 on cultured cells. To determine the pathogenicity of CTNCEC25, suckling mice, adult mice, guinea pigs and rabbits were inoculated with CTNCEC25 via the intracerebral route and their survival monitored every day. Furthermore, the CTNCEC25 strain was passed serially in CECs for 20 passages and then 3 passages in the brains of suckling mice to determine phenotypic stability. CTNCEC25 achieved similar growth kinetics in Vero cells and N2a cells compared with parental CTN-1, but CTNCEC25 replicated more efficiently in CECs than the CTN-1 strain with a titer 72 h after infection reaching 10(7.5-7.6) FFU/mL, which was significantly higher than the 10(5.8) FFU/mL achieved by its parental strain, CTN-1. Moreover, CTNCEC25 induced apparent CPE in Vero cells, CECs and N2a cells. Analyses of intracerebral inoculation demonstrated that CTNCEC25 was attenuated profoundly in adult mice and was completely apathogenic to guinea pigs and rabbits, though it caused death in suckling mice. The CTNCEC25 strain proliferated steadily after serial passage in CECs and the brains of suckling mice, and remained avirulent in adult mice. These results suggest that CTNCEC25 is a highly attenuated and genetically stable strain of the rabies virus. CTNCEC25 replicated stably and efficiently in cultured cells and achieved high titers, so it could be a promising and safe vaccine strain for rabies prevention in China.
Animals ; Cell Line ; Cercopithecus aethiops ; China ; Guinea Pigs ; Humans ; Mice ; Rabbits ; Rabies ; virology ; Rabies virus ; genetics ; growth & development ; pathogenicity ; Serial Passage ; Viral Vaccines ; adverse effects ; genetics ; Virulence

Objective To evaluate the clinical effectiveness of double balloon dilation in percutaneous kyphonplasty on curing vertebral?compression fractures. Methods From January 2009 to September 2013, 84 patients (94 vertebral bodies) with vertebral compression were treated by percutaneous kyphonplasty. All were fresh fractures and were injured or obvious low back pain 1 month, accompanied by local tenderness, kowtow attack painful, lumbar mobility, but no lower extremity injury numbness, activities and defecation disorders. After randomization, the double balloon dilation in percutaneous kyphonplasty method was used to treat 44 patients (49 vertebrae). In the process of performing percutaneous unilateral pedicle puncture and balloon dilata?tion of the vertebral body, the balloon has been moved some distance in the vertebral body. Then completed the perfusion of bone cement, vertebral body forming. 40 cases (45 vertebrae) were used conventional unilateral percutaneous kyphonplasty to vertebtal compression fractures. Recorded the operation time, amount of bleeding, bone?cement injection volume. Used visual analogue scale (VAS), the height of the vertebral body and Cobb angle to evaluated the curative effect. Results All 84 patients completed the operation, follow?up time was 22 months (18-24 months). In two mobile open expansion group, the operation time was about 48 min. The amount of bleeding was 8-15 ml. The average bone?cement injection volume was 5.1 ml. No patients quit the study and no bone cement?leakage cases or other side effects were observed , and no clinical accidents occurred. In a single stretching group, 40 cases (45 vertebrae) completed conventional vertebroplasty, the time of 44 min, bone cement average injection rate 3.2 ml, bleeding 10-15 ml. In two mobile open expansion group, the VAS score was 8.5 points, the height of the vertebral body height was 2.1cm, and the Cobb angle was 34°. After operation, the VAS score 2.9 points, the height of the vertebral body 2.8 cm, and Cobb 20° . In the other group, the pain was significantly relieved and the relief was satisfactor after operation.Vertebral height of 2 cm turned to the last follow?up of 2.4 cm. The Cobb angle was 32°, and the last follow?up was 27°. The VAS score, operation time and bleeding volume of the two groups were not statistically significant, and a statistically significant difference of the average bone ce?ment injection volume, postoperative vertebral height and cobb angle improved with statistical significance. A single open group were 2 cases of bone cement leakage and leakage, 1 cases of bone cement tail, the complication rate was 5.6%. Conclusion The application of double balloon dilation in percutaneous kyphonplasty to vertebral?compression fractures improve relocation of verte?bral compression fractures, increase recovery of vertebral height, and more effectively strengthen and stiffen pathological vertebral bodies, while improving kyphosis. Moreover, it can reduce pressure during bone?cement injections, minimizing the chance of over?flow and leakage, as well as the related side effects, but it will also result in an increase of bone cement?injection volume.

Objective To discuss the diagnosis and treatment of splenic harmatoma(SH). Methods The clincial data of three cases of SH treated in our hospital from January 1997 to December 2007 were collected,and the other 17 cases which were published from January 1997 to December 2007 in the Chinese biological and medical lit-erature database were reviewed. The clinical manifestation,pathological findings,diagnosis and treatment of these 20 patients were analyzed. Results The main symptoms were abdominal pain or discomfort and abdominal mass in 13 cases(13/20). All 20 patients underwent surgical treatment with good recovery without severe complications and re-currence. Conclusions The definite diagnosis of SH depends on postoperative pathological findings. Complete surgi-cal resection is the best treatment for SH with favourable prognosis.