Objective Through detecting CD4+CD25+ T regulatory cells(Treg)in the peripheral blood in children suffering autoimmune diseases and normal controls to learn about the changes of Tregs during the diseases and to acquire some references for clinical diagnosis and treatment.Methods The data were reviewed for CD4+CD25+ Treg cells of the 93 children diagnosed as pediatric autoimmune disease in Children′s Hospital Affiliated to Capital Institute of Pediatrics from Nov.2007 to Jun.2008.Thirty-five normal children in the contempora-neous physical examination were selected as the control group.The percentage of CD4+CD25+ Treg cells and CD4+ T cells to the total T cells were determined by flow cytometric method.Data of the JRA group(22 cases),SLE group(12 cases) and HSP group(12 cases),which were the first three according to the number of cases,were respectively compared with the controls.Independent-samples t test was performed for a statistic analysis with SPSS 11.5 software.Results 1.The percentages of CD4+CD25+ Treg cells to the total T cells and CD4+ T cells in the autoimmune diseases children[(6.14?3.21)% and(21.85?11.68)%,respectively] were both higher than those in the control group[(3.68?1.02)% and(12.83?3.61)%,respectively Pa

Objective To investigate the clinical cross infections of myeoplasma pneumoniae(MP)and other viruses in children,providing a reference for the diagnosis and treatment of respiratory disease.Methods Serum specimens of the children hospitalized with fever,respiratory symptom besides positive results of MP-Ab IgM detection were collected.And several common viruses popular in children were investigated within the specimens collected by ELISA kits or indirect immunofluorescence.Results (1)The PCT levels of 385 cases(81.7%)appear to be under 0.5 ng/ml.(2)In the 514 cases detected for Cox-IgG and Cox-IgM,the positive rates are respectively 40.3% and 35.6%.(3)2 cases(0.8%)appear to be irdluenza B virus positive.And the positive rates of parainfluenza virusl,2 and 3 are 0.8%,0,and 9%.4,84 cases(11.8%)are positive for EB-IgM and 451 cages(63.6%)positive for EB-IgG.Conclusion Cross infections rarely occur between MP and common respiratory viruses in Children.The cross-infection rate between Cox-virus and MP iS up to 35.6%.

OBJECTIVETo observe the clinical efficiency of curettage combining circumcision on giant condyloma acuminata(CA) in male external genitalia, the relationship between recurrence and curettage depth, the possibility of HPV infection in PBMC after operation.

METHODSCurettage combining circumcision was carried out on 50 cases with CA. The removed wart and wound surface tissues were examined under light microcope and for HPV-DNA detection by PCR. HPV-DNA was detected in PBMC during < or =1 and > or =2 weeks after operation.

RESULTS(1) 46 cases were cured completely after one treating (once the cure rate is 92%), 4 cases twice(twice the cure rate is 8%); (2) The tissues were proved to be HPV-DNA positive by PCR amplification and CA relapse occurred; (3) HPV-DNA was detected in PBMC only within the week after curettage in the 19 cases.

CONCLUSIONS(1) The clinical efficiency of curettage combining circumcision on giant condyloma acuminata in male external genitalia is sure; (2) Cure rate and relapse rate are related with curettage depth; (3) Transient positive HPV-DNA in PBMC may be detected.

Adolescent ; Adult ; Aged ; Circumcision, Male ; Condylomata Acuminata ; surgery ; Curettage ; methods ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo investigate the effect of Wenjingtong Composita (WJTC) on blood glucose and advanced glycosylation end products (AGEs) in sciatic nerve of streptozotocin (STZ)-induced diabetic rats.

METHODSSTZ-induced diabetic rats were randomized to WJTC prevention group and WJTC treatment group. The levels of blood glucose and AGEs in sciatic nerve of the animals were checked after 12 weeks treatment and compared with that of aminoguanidine (AG) treatment group.

RESULTSBlood glucose level in the WJTC prevention and treatment group, and AGEs in sciatic nerve of the WJTC prevention group and the AG group were lower than those of the non-treated group (P < 0.01). Blood glucose level in the AG group was higher than that in the WJTC prevention group (P < 0.05), but was not significantly different from that in the non-treated group (P > 0.05).

CONCLUSIONWJTC might prevent diabetic peripheral neuropathy by decreasing blood glucose and inhibiting AGEs formation in sciatic nerve in STZ-induced diabetic rats.

Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Experimental ; blood ; metabolism ; Diabetic Neuropathies ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Glycosylation ; drug effects ; Guanidines ; pharmacology ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Sciatic Nerve ; metabolism ; Sciatic Neuropathy ; metabolism

BackgroundThe quest to look for seed cells is a hot spot of cornea transplant research in solving the problem of the lack of donor. Bone marrow mesenchymal stem cells(BMSCs) have been successfully induced into retinal ganglion cells(RGCs) in vivo,but the successful induction of BMSCs into corneal endothelial cells has not been reported.Objective This experiment was to study the transplantation of BMSCs on corneal endothelial surface using the splitting Descemet's membrane. MethodsFour healthy adult rhesus monkeys were divided into the experimental group ( 3 monkeys) and control group ( 1 monkey). Mesenchymal stem cells (MSCs) were isolated from bone marrow by density gradient centrifugation combined with adhering means. The cultured cells were identified by flow cytometry and its ability to differentiate was determined by allowing them to differentiate into adipocytes in vitro and labeled by 5-bromodeoxyuridine ( BrdU ) for subsequent identification. Corneal grafts of 7 mm in size with tearing of the Descemet' s membrane were prepared in the experimental group and control group. After labeling by 5-bromodeoxyuridine( BrdU ) ,cultured cells were transplanted onto the endothelial surface of cornea grafts in the experimental group, but no cultured cells were seeded in the graft of the control group. The corneal grafts were then sutured in situ, and were removed 1,2 or 3 months after operation to examine the distribution and connection between transplanted cells and their morphologic changes under the electron microscope. Results High purity MSCs were harvested by density gradient centrifugation combined with adhering method. Cultured cells reached confluency after 12 to 16 days, presenting with a spindle shape and parallel or swirling arrangement. Flow cytometry analysis showed that 94.26％ of cells were positive for CD29,7. 51％ for CD34 and 4. 02％ for CD45. Larger nuclei filled with plastosomes, golgiosomes and rough endoplasmic reticula were found on the graft under the transmission electron microscope( TEM ). After 3 weeks, MSCs were differentiated into adipocytes where Oil Red O staining resulted in an orange-red staining in the cytoplasm and blue staining in the nuclei. The transplanted cells attached loosely on the endothelial surface of the corneal graft and came in contact with each other in one month. The shape of the cells appeared as spindle-shaped and polygonal after 2 months and became tightly packed after 3 months. The positive cells retained the BrdU label and presented with brown nuclei. No endothelia cells grew in the cornea graft in the control group, with an absence of BrdU labeling. Conclusions Mesenchymal stem cells can be transplanted onto the corneal endothelial surface successfully and form a monolayer using the centrifugation method, and present with good survival and proliferation ability.

OBJECTIVETo investigate the clinical features of Epstein-Barr virus-related hemophagocytic lymphohistiocytosis (EBV-HLH), to analysis the outcome of HLH-2004 protocol, and to explore the prognostic factors in EBV-HLH patients.

METHODSThe clinical features at onset and outcome of HLH-2004 protocol from 83 pediatric patients with EBV-HLH enrolled from January 2006 to December 2009 in our hospital were analyzed retrospectively. Univariate and multivariate COX regression analysis were used to identify statistically significant prognostic factors.

RESULTS(1) Among the 83 patients, 45 were males and 38 were females. The age of onset ranged from 6 months to 14 years 4 months. 44 patients were treated with HLH-2004, and 3-year overall survival (OS) was (55.8 ± 7.9)%. (2) The most common clinical features of EBV-HLH included high fever, cytopenia, hepatosplenomegaly, and coagulopathy; The respiratory symptoms, angina phlogistic, skin rashes, neurologic abnormality were rare. 97.3% of patients showed an elevation of serum ferritin, liver dysfunction and lipid metabolism disorders was found in most of EBV-HLH patients. 89.0% of patient had hemophagocytosis in bone marrow at diagnosis of EBV-HLH. (3) COX regression analysis revealed that anemia degree, serum albumin < 30 g/L, CD4:CD8 abnormity, NK cell < 3%, treatment protocol were related with the prognosis significantly (P < 0.05).

CONCLUSIONEBV-HLH in pediatric patients has severe clinical feature and poor prognosis. HLH-2004 protocol is an effective treatment for patients with EBV-HLH. Symptomatic treatment can't rescue the patients of EBV-HLH.

Adolescent ; Child ; Child, Preschool ; Epstein-Barr Virus Infections ; drug therapy ; Female ; Herpesvirus 4, Human ; Humans ; Infant ; Killer Cells, Natural ; Lymphohistiocytosis, Hemophagocytic ; drug therapy ; virology ; Male ; Prognosis ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo explore the mechanism of Shangke Jiegu Tablet (SJT)in repairing the mandibular defect.

METHODSTotally 72 healthy male New Zealand rabbits were randomly divided into the normal control group (n = 24), the model group (n = 24), and the SJT group (n = 24). Then the mandibular defect model was established. Animals in the normal control group and the model group were fed with normal forage, while those in the SJT group were fed with SJT forage. On the day 7, 14, 28, and 56 after model establishment, 6 rabbits were killed in each group. The bone was collected from the mandibular defect. The gene expressions of osteoprotegerin (OPG) and osteoprotegerin ligand (OPGL) were detected by means of RT-PCR. The positive dyeing strength and area of the bone tissue were detected by means of immunohistochemical technique.

RESULTSCompared with the normal control group, the degree of OPGmRNA expression was remarkably up-regulated on day 7 after model establishment (P < 0.05) and the degree of OPGLmRNA expression was remarkably up-regulated on day 14 after model establishment (P < 0.05) in the model group. Compared with the model group, the degree of OPGmRNA expression was remarkably up-regulated (P < 0.05), and the positive dyeing strength and area of bone tissue were stronger and broader on day 14, 28, and 56 after model establishment in the SJT group. The degree of OPGLmRNA expression was remarkably down-regulated (P < 0.05), and the positive dyeing strength and area of bone tissue were weaker and smaller on day 14 after model establishment in the SJT group. The ratio of OPGmRNA/OPGLmRNA was remarkably up-regulated on day 14, 28, and 56 after model establishment (P < 0.05).

CONCLUSIONThe effect mechanism of promoting mandibular defect repairing by SJT may be correlated to regulating the expressions of OPGmRNA and OPGLmRNA.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression ; drug effects ; Ligands ; Male ; Mandibular Injuries ; genetics ; metabolism ; Osteoprotegerin ; metabolism ; RANK Ligand ; metabolism ; Rabbits ; Wound Healing ; drug effects

OBJECTIVETo explore the potential of iodized linoleic acid (ILA) and its 5-fluoro-deoxyuridine ester (IFU) to inhibit hepatocellular carcinoma (HCC) cells in vitro and tumors in vivo.

METHODSILA and its constituent component IFU were chemically synthesized, purified, and confirmed by 1H-NMR. The HCC cell lines, QGY-7703 (5-fluorouracil (5-FU) treatment sensitive) and SMMC-7721 (5-FU resistant), were treated with ILA, IFU, 5-FU, or traditional lipiodol for 72 hours. Survival rates of the treated cells were assessed by the methyl thiazolyl tetrazolium method, and used to calculate the IC50 and IC90. In addition, thirty nude mice were subcutaneously inoculated with SMMC-7721 cells and randomly divided two weeks later into four treatment groups (n = 6 each) for intra-tumoral injection of ILA, IFU, 5-FU, lipiodol or DMSO (controls). The rate of tumor inhibition (RTI) was calculated for each group at week 4 after treatment.

RESULTSFor the cultured SMMC-7721 cells, the inhibitory concentrations for ILA, IFU, and 5-FU were: IC50: 134.38 mumol/L, 17.55 mumol/L, and 7.38 mumol/L; IC90: 192.88 mumol/L, 97.63 mumol/L, and more than 200 mumol/L. For the cultured QGY-7703 cells, the inhibitory concentrations for ILA, IFU, and 5-FU were: IC50: 109.55 mumol/L, 44.79 mumol/L, and 98.06 mumol/L; IC90: all, more than 200 mumol/L. In both cell types, the IC50 of lipiodol was more than 400 mumol/L. Compared with the RTI of the control mice (100%), the RTI of ILA-treated mice was 31.9% (t = 2.37, P less than 0.05), of IFU-treated mice was 56.9% (t = 4.91, P less than 0.01), and of 5-FU-treated mice was 31.0% (t = 2.59, P less than 0.05). The RTI of IFU was significantly stronger than that of either ILA or 5-FU (P less than 0.05). The lipiodol treatment showed no inhibition effect on tumors (P more than 0.05).

CONCLUSIONILA and IFU can effectively inhibit the growth of HCC cells in vitro and tumors in vivo. Furthermore, IFU outperforms ILA in inhibiting HCC growth.

Animals ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Fluorouracil ; pharmacology ; Humans ; Inhibitory Concentration 50 ; Linoleic Acid ; pharmacology ; Liver Neoplasms ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Xenograft Model Antitumor Assays

OBJECTIVETo develop a LC-MS method for the determination of senkyunolide I (SI) in rat plasma, in order to observe whether there is significant change in the pharmacokinetics parameters of complex prescriptions of Huoluoxiaolingdan (HLXL) and single herbal extracts from Ligusticum chuanxiong Hort. in rats, and assess the effect of other components in HLXL on the pharmacokinetics of SI.

METHODTwelve male Sprague-Dawley (SD) rats were randomly divided into two groups, and orally administered with extract from HLXL and L. chuanxiong (both equal to SI 4.53 mg x kg(-1)). Their blood was collected at different time points for LC-MS, in order to detect the plasma concentration of SI. The pharmacokinetic parameters of SI were calculated by DAS 2.0 software. SPSS 16.0 software was used for independent-sample T-test and Nonparametric T-test.

RESULTA linear relationship of SI ranged from 6.750 to 675.0 microg x L(-1), and with the lowest limit of detection being 6.750 microg L(-1). Both of the plasma concentration-time curves of SI were fitted with the two-compartment model for extract of HLXL and L. chuanxiong. The detected AUC and Cmax of SI showed significant difference, with no significant difference in other parameters.

CONCLUSIONThe LC-MS determination method established in this experiment was so exclusive, accurate and sensitive that it is suitable for pharmacokinetic studies on extracts of HLXL and SI from L. chuanxion. The experiment results show that other ingredients of HLXL have noticeable effect on the absorption of SI in rat plasma.

Administration, Oral ; Animals ; Area Under Curve ; Benzofurans ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Male ; Metabolic Clearance Rate ; Random Allocation ; Rats ; Rats, Sprague-Dawley

AIMTo investigate human epidermal growth factor receptor type 2 (HER2) protein expression and gene amplification in Chinese metastatic prostate cancer patients and their potential value as prognostic factors.

METHODSImmunohistochemistry (IHC) was performed to investigate HER2 protein expression in prostate biopsy specimens from 104 Chinese metastatic prostate cancer patients. After 3-11 months of hormonal therapy, 12 patients underwent transurethral resection of the prostate (TURP). HER2 protein expression of TURP specimens was compared with that of the original biopsy specimens. Of these, 10 biopsy and 4 TURP specimens with HER2 IHC staining scores >or=2+ were investigated for HER2 gene amplification status by fluorescent in situ hybridization (FISH).

RESULTSOf the 104 prostate biopsy specimens, HER2 protein expression was 0, 1+, 2+ and 3+ in 49 (47.1%), 45 (43.3%), 8 (7.7%) and 2 (1.9%) cases, respectively. There was a significant association between HER2 expression and Gleason score (P = 0.026). HER2 protein expression of prostate cancer tissues increased in 33.3% of patients after hormonal therapy. None of the 14 specimens with HER2 IHC scores >or= 2+ showed HER2 gene amplification. Patients with HER2 scores >or= 2+ had a significantly higher chance of dying from prostate cancer than those with HER2 scores of 0 (P = 0.004) and 1+ (P = 0.034). Multivariate Cox regression analysis showed that HER2 protein expression intensity was an independent predictor of cancer-related death (P = 0.039).

CONCLUSIONAn HER2 IHC score >or= 2+ should be defined as HER2 protein overexpression in prostate cancer. Overexpression of HER2 protein in cancer tissue might suggest an increased risk of dying from prostate cancer. HER2 protein expression increases in some individual patients after hormonal therapy.

Aged ; Antineoplastic Agents, Hormonal ; therapeutic use ; Asian Continental Ancestry Group ; genetics ; statistics & numerical data ; Biopsy ; China ; epidemiology ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Prognosis ; Proportional Hazards Models ; Prostatic Neoplasms ; drug therapy ; genetics ; mortality ; secondary ; Receptor, ErbB-2 ; genetics ; metabolism ; Risk Factors