Objective To evaluate the diagnostic value of detection of IgM antibodies to EV71-infection patients,and compared characterisation of RT-PCR,IgM capture ELISA and neutralization test.Methods Virus RNA,neutralization titer and IgM antibody in 115 EV71-infection patients were detected by EV71 real-time RT-PCR kit( EV71-PCR kit),neutralization test,and EV71 IgM-capture ELISA kit (EV71-IgM kit),respectively.Results Using EV71-IgM kit,the detection rate was 80.9％ (93/115,95％ CI:72.5-87.6) among the 115 EV71-infection patients,and was 2.6％ among the 228 healthy children.Simultaneously,sera collected after 1-2 day of disease onset showed an IgM positivity of 70.4％ (38/ 54).The positive rate of EV71-PCR among these patients was 82.6％ (95/115,95％ CI:74.4-89.0),so there was no statistically significant differences between it and EV71-IgM kit.In addition,the detection rate in EV71-infection patients could increase to 92.2％ by combined detection of EV71-PCR and EV71-IgM kit.Conclusion EV71-IgM kit was a rapid and valuable way for the early diagnosis of EV71 infection,and could significantly improve detection rate for EV71 infection by combining with EV71-PCR kit.

We revisited the "Xiao Chaihu Decoction event (XCHDE)" occurred in late 1980s in Japan and the Bupleuri Radix related adverse drug reaction (ADR) reports in China After careful review, comparison, analysis and evaluation, we think the interstitial pneumonitis, drug induced Liver injury (DILI) and other severe adverse drug envents (ADEs) including death happened in Japan is probably results from multiple factors, including combinatory use of XCHDE with interferon, Kampo usage under modern medicine theory guidance, and use of XCHD on the basis of disease diagnosis instead of traditional Chinese syndrome complex differentiation. There are less ADE case reports related to XCHD preparation in China compared to Japan, mostly manifest with hypersensitivity responses of skin and perfuse perspiration. The symptoms of Radix Bupleuri injection related ADEs mainly manifest hypersensitivity-like response, 2 cases of intravenous infusion instead of intramuscular injection developed hypokalemia and renal failure. One case died from severe hypersensitivity shock. In Chinese literatures, there is no report of the interstitial pneumonitis and DILI associated with XCHDG in Japan. So far, there is no voluntary monitoring data and large sample clinical research data available. The author elaborated the classification of "reevaluation" and clarified "re-evaluation upon events" included the reaction to the suspected safety and efficacy events. Based on the current status of the clinical research on the Radix Bupleuri preparations, the author points out that post-marketing "re-evaluation upon suspected event" is not only a necessity of continuous evaluation of the safety, efficacy of drugs, it is also a necessity for providing objective clinical research data to share with the international and domestic drug administrations in the risk-benefit evaluation. It is also the unavoidable pathway to culture and push the excellent species and famous brands of TCM to the international market, in that it enhances the pharmaceutical manufactures the ability to control enterprise risk and eliminate the negative impact on traditional Chinese medicine. With the widely used and ADR report concentrated Radix Bupleuri injection as an example, the author provide a new approach to post-marketing clinical research.
Bupleurum ; chemistry ; Drug Compounding ; Drug-Related Side Effects and Adverse Reactions ; etiology ; Drugs, Chinese Herbal ; adverse effects ; Humans ; Product Surveillance, Postmarketing ; methods ; Safety

ObjectiveTo investigate norovirus infection status and indentify its epidemiological characteristics and genotype distribution in infantile viral diarrhea in Jiangsu.MethodsFour hundred and ninety-eight fecal specimens of infantile virus diarrhea cases were collected from Suzhou Children's hospital and Nanjing Children's hospital in 2010.Norovirus genegroup were detected by real-time RT-PCR,and genetype were determined by sequence analysis.Results Among all fecal specimens,2 (0.4％) cases were positive for norovirus G Ⅰ,and 190 (38％) cases for G Ⅱ.Nucleotide sequence analysis revealed that in the 2 samples for G Ⅰ,one strain was G Ⅰ 1 and another was G Ⅰ 3.Twenty-one strains were belonged to G Ⅱ 4 and 2 strains were G Ⅱ 3 in the 23 samples for G Ⅱ.ConclusionAs one of the most important pathogens causing infantile viral diarrhea in Jiangsu province,subtype G Ⅱ 4 was the main epidemic strain of norovirus,meanwhile other genotypes also existed.

OBJECTIVETo analyze and summarize the clinical and pathological characteristics of intracranial haemangioblastoma and to improve surgery effect.

METHODSeventy-two patients with intracranial haemangioblastoma who were proven by operation and pathology from 1970 to 1988 were analyzed retrospectively.

RESULTSIntracranial haemangioblastoma tends to occur in the hemisphere of cerebellum (83 tumours, 87%) and the age of them ranged from 20 to 40 years (47 cases, 58.3%) mostly. The ratio of men (46 cases) was higher than women (26 cases). The diagnosis of the disease depends on CT and MR substantive haemangioblastoma. The most effective and reliable treatment of intracranial haemangioblastoma is surgical resection.

CONCLUSIONIntracranial haemangioblastoma is benign tumour which can be, cured by total surgical resection. The key recurrence factors include the young age of initial onset, mistaken exploration and incomplete extirpation of tumour.

Adolescent ; Adult ; Brain Neoplasms ; pathology ; surgery ; Child ; Female ; Hemangioblastoma ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo examine the correlation between p65 expression and the metastasis and prognosis in tongue carcinoma.

METHODSThe immunohistochemical expression of p65 was examined in 50 cases of tongue carcinoma, and the clinical and histopathologic records were retrospectively analyzed.

RESULTSOverexpression of p65 in cytoplasma and nucleus was found in 64% cases of tongue carcinoma. There was significant difference of p65 expression among different TNM stages, and the higher expression rate of p65 was found in higher stages. Overexpression of p65 was also related to lymph node metastasis. Significantly greater increase in expression levels of p65 was found in the metastatic cases (n = 22) compared to the nonmetastatic cases (n = 10). However, there was no significant difference of p65 expression among different histopathologic grading. The 5-year survival rate of p65 positive and negative group was 59.3% and 88.9% respectively, and there was significant difference in Kaplan-Meier survival curve (chi2 = 5.6, P < 0.05).

CONCLUSIONThe results suggest that the expression of p65 was important to predict the metastasis and prognosis in tongue carcinoma.

Adult ; Aged ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Tongue Neoplasms ; Transcription Factor RelA

OBJECTIVETo examine the relationship between NF-kappaB signal transduction way and the sensitivity of Tca8113 carcinoma cell to Pingyangmycin chemotherapy.

METHODS2 mg/L antisense p65 oligodeoxynucle-otides (5'-GAACAGTTCGTCCATGGCCG-3') was transfected into Tca8113 cells through Lepofectin vectin, then the cells were treated with 8 mg/L Pingyangmycin. After 3 h and 6 h, the expression of p65 in nucleus was studied with immunohistochemical methods and Western blot analyses. After 48 h, the inhibitory rate of cell growth was detected with MTT assay.

RESULTSThe NF-kappaB/p65 signal transduction way in Tca8113 cells was activated after treated with Pingyangmycin. But the transfection of antisense p65 oligodeoxynucle-otides inhibited the activation of the signal transduction, the p65 expression in nucleus was decreased significantly (P < 0.05) at 6 h, and the inhibitory rate of cell growth was increased significantly (P < 0.05) at 48 h.

CONCLUSIONThe results suggested the chemotherapeutic sensitivity of Tca8113 carcinoma cell to Pingyangmycin was improved by the inhibition of NF-kappaB/p65 signal transduction.

Bleomycin ; analogs & derivatives ; Cell Line, Tumor ; Cell Proliferation ; Humans ; NF-kappa B ; Signal Transduction ; Tongue Neoplasms ; Transcription Factor RelA ; Transfection

Using the character of natural aggregation of CHO cells, and an ultrasonic and sedimentation column combined perfusion system to promote cells aggregation and retention into bioreactor,recombinant CHO cell strain MK3-A2 was cultured,which could secrete rhTNK-tPA, by a serum-free perfusion culture system. The culture periods in this two experiments were as long as 77 and 110 days respectively. The cells density reached 2?107 cells /ml. The average volumetric productivity of rhTNK-tPA was 89 mg/L?d, and the highest one was 216mg/L?d.The cells aggregation rate was approximately 90%, and the diameters of most of them were 285～570?m. During the perfusion culture the cells retention rate almost kept in 95% and the viability of cells was more than 85%.Thus, it means that aggregation culture with such perfusion system could be used to scale up produce biopharmaceuticals instead of microcarrier culture system.

BACKGROUNDTransplantation of mensenchymal stem cells (MSCs) has been proposed as a promising way for tissue engineering. However, the application of MSCs for transplantation will undergo apoptosis due to the extremely harsh microenvironment such as excessive inflammation. Apigenin (API) has been reported to protect cells against inflammatory damage and cell death by exhibiting anti-inflammatory and anti-oxidative capacity. Here we investigated the modulatory effects of API in lipopolysaccharide (LPS)-mediated inflammation and apoptosis of MSCs, and further defined the underlying mechanism.

METHODSEffects of different concentrations of API (0, 5, 10, 20, 40 and 80 µmol/L) for 24 hours, and LPS (0, 0.5 and 5.0 µg/ml) for 6 hours and 24 hours on MSCs viability were assayed by MTT. Based on this, MSCs were pretreated with different concentrations of API (0 - 40 µmol/L) at the indicated times (6, 12 and 24 hours) followed by exposure to 5 µg/ml LPS for 24 hours. MTT, phase-contrast microscopy, annexinV/propidium iodide (PI) double stain flow cytometry (FCM) and Hoechst staining were applied to explore the effects of API on MSCs induced by 5 µg/ml LPS for 24 hours. In addition, reverse-transcription polymerase chain reaction (RT-PCR) was applied to detect the mRNA expression of pro-inflammatory factors including cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), nuclear factor-kappa B (NF-κB), pro-apoptotic gene caspase-3, Bad, and anti-apoptotic gene Bcl-2. Moreover, AutoDock software was used to imitate the docking score of API and vitamin D receptor (VDR). In parallel, Western blotting and RT-PCR were used to investigate protein and mRNA expression of VDR.

RESULTSMSCs stimulated with LPS 5 µg/ml for 24 hours was used as a model of apoptosis induced by over inflammatory stimulus. API (0 - 40 µmol/L) had non-toxic effect on MSCs; however, it could decrease mRNA expression of COX-2, iNOS and NF-κB at different time points in MSCs induced by LPS, except for API at the concentration of 5 µmol/L.

RESULTSfrom phase-contrast microscopy, MTT, Hoechst staining and AnnexinV/PI double stain FCM demonstrated that with the increasing concentrations of API and extension of administrating time, significant morphological changes of MSCs occurred, viability of cells was strongly inhibited, and meanwhile, apoptosis of LPS-administrated MSCs was exacerbated, compared with LPS individual group. In addition, API promoted caspase-3, Bad mRNA expression and inhibited Bcl-2 mRNA expression in a time-dependent and concentration- dependent manner. Further study found that pro-apoptosis effect of API was related to suppress VDR expression.

CONCLUSIONSAPI could inhibit the expression of inducible inflammatory factors, therefore exert the strong anti-inflammatory function. However, API could not protect MSC apoptosis induced by LPS but amplified the apoptosis. The apoptosis is related to Bad/Bcl-2 increasing and caspase-3 activation, which is mediated through suppressing VDR expression.

Animals ; Apigenin ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Cell Survival ; drug effects ; Cells, Cultured ; Flow Cytometry ; Lipopolysaccharides ; pharmacology ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Calcitriol ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo examine whether transforming growth factor-β (TGF-β) pathway and adaptive T cell immunity play roles in the anti-atherosclerotic effects of pioglitazone (PIO) in ApoEmice.

METHODSApoEmice with atherosclerosis induced by high-fat feeding were treated daily with PIO (20 mg/kg) or vehicle for 8 weeks. The protein expressions of TGF-β pathway in the atheromatous lesions of the aorta and the percentages of IFN-γand Foxp3cells in the spleen of the mice were examined with immunohistochemical staining. In the in vitro experiment, primary cultured splenocytes were stimulated with oxidized low-density lipoproteins (oxLDL) and treated with PIO either alone or in combination with the PPARγ antagonist GW9662, after which the changes in percentages of CD4IFN-γcells and CD4CD25Foxp3cells were analyzed with flow cytometry.

RESULTSPIO treatment of ApoEmice with high-fat feeding significantly attenuated the progression of atheromatous lesions (P<0.05) and resulted in increased expressions of TGFβ1 (P<0.01), TGFβRII (P<0.05), and p-Smad3 (P<0.05) and a decreased expression of Smad7 (P<0.05) in the lesions. PIO treatment also led to decreased percentage of IFN-γcells (P<0.05) and increased percentage of Foxp3cells (P<0.01) in the spleen of the mice. In primary cultured splenocytes, PIO treatment caused significant down-regulation of IFN-γ mRNA (P<0.05) and up-regulation of Foxp3 mRNA (P<0.05) and obviously increased the percentages of CD4IFN-γcells (P<0.05) and CD4CD25Foxp3(P<0.05); the effects of PIO on CD4IFN-γand CD4CD25Foxp3cells were abolished by treatment of the cells with GW9662.

CONCLUSIONThe anti-atherosclerotic effect of PIO is probably mediated by the TGF-β/Smad signaling pathway and PPAR-γ-dependent modulation of Th1/Treg population.

OBJECTIVETo analyze the etiological and epidemiological characteristics of hand-foot-and-mouth disease (HFMD) seen in Jiangsu province from 2008 to 2010, and provide evidence for its prevention and control.

METHODSBased on the requirement of supervision program of HFMD, surveillance and report were done according to National Disease Supervision Information Management System. Descriptive epidemiological method, performed between 2008 and 2010, was used to analyze the time, region and population distribution and results of etiologic analysis of HFMD. Nucleic acid of enterovirus (EV) genome was detected by real-time RT-PCR.

RESULTSThe average incidence rate of HFMD was 86.70 per million between 2008 and 2010, the peak incidence occurred in April to July. There were significant differences among the incidence in different districts (P<0.05), and the highest incidence was seen in the densely inhabited southern areas of Jiangsu province. Most of the cases were infants and children aged less than 5 years. The number of male cases (2008: 17,008, 2009: 48 768, 2010: 50,231) was much larger than that of the female cases 2008: 9662, 2009: 29 151, 2010: 30,655. The HFMD cases with mild symptoms were caused mainly by enterovirus 71 (EV71) and coxsackievirus A16 (Cox A16) and there was difference among different years. The severe HFMD cases and deaths were mainly caused by EV71 infection.

CONCLUSIONSThe epidemiologic characteristics of HFMD in Jiangsu province from 2008 to 2010 had close relationship with season, population and region. The mild cases of HFMD were mainly infected with EV71 and Cox A16. However, EV71 illness seemed to be more severe and had significantly greater frequency of serious complications and fatality than the illness caused by Cox A16.

Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; Enterovirus A, Human ; pathogenicity ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; virology ; Humans ; Infant ; Male