Animals ; Forecasting ; Herpesvirus 2, Gallid ; genetics ; physiology ; Marek Disease ; genetics ; metabolism ; virology ; MicroRNAs ; metabolism

OBJECTIVE: To explore the cross-talk between extracellular signal-regulated kinase (ERK) and nuclear factor (NF-kappaB) signal transduction pathways in the hepatocytes expressing hepatitis C virus nonstructural protein 3 (HCV NS3). METHODS: A cell line QSG7701/NS3, which stably expressed HCV NS3 protein, was constructed by transfecting plasmid pcDNA3.1-NS3 into a human immortalized hepatocyte line QSG7701. Before and after QSG7701/NS3 cells were treated by MEK inhibitor PD98059, the phosphorylation level of ERK and the expression of cyclin D1 protein were detected by Western blot; the DNA binding activities of activator protein 1 (AP-1) and NF-kappaB were evaluated with electrophoretic mobility shift assay (EMSA). Cell cycles were measured by flow cytometry (FCM); the effects of PD98059 on the cell proliferation were determined by MTT assay. RESULTS: HCV NS3 protein could up-regulate the phosphorylation of ERK and the expression level of cyclin D1 in QSG7701 cells. PD98059 could inhibit the cell proliferation mediated by HCV NS3 protein, down-regulate the activities of AP-1 and NF-kappaB, and suppress the expression of cyclin D1. CONCLUSION The inhibition of ERK can suppress the DNA binding activity of NF-kappaB and the cell proliferation mediated by HCV NS3 protein in a dose-dependent manner. There may be a cross-talk between ERK and NF-kappaB signal transduction pathways, which may exert synergistic action on the proliferation and malignant transformation of hepatocytes.
Blotting, Western ; Cell Cycle ; drug effects ; Cell Line ; Cell Proliferation ; drug effects ; Cyclin D1 ; metabolism ; Dose-Response Relationship, Drug ; Extracellular Signal-Regulated MAP Kinases ; antagonists & inhibitors ; metabolism ; Flavonoids ; pharmacology ; Hepatocytes ; cytology ; metabolism ; Humans ; NF-kappa B ; metabolism ; Phosphorylation ; drug effects ; Signal Transduction ; drug effects ; physiology ; Viral Nonstructural Proteins ; genetics ; metabolism

OBJECTIVETo explore the effect of Qiling Decoction (QD) combined highly active antiretroviral treatment (HAART) on expression levels of peripheral blood Th17 and Treg cells in HIV/AIDS patients.

METHODSTotally 55 HIV/AIDS patients were randomly assigned to the treatment group (28 cases) and the combination group (27 cases). Besides, 21 HIV negative patients were recruited as the healthy control group. Those in the treatment group received HARRT alone, while those in the combination group received HAART combined QD. The observation lasted for 24 weeks. Meanwhile, according to peripheral blood CD4+ T cell counts before treatment, HIV/AIDS patients were assigned to three subgroups. For patients in subgroup 1, 1 cells/microL < CD4+ T cell counts < or = 100 cells/microL; For patients in subgroup 2, 101 cells/microL < CD4+ T cell counts < or = 200 cells/lL; For patients in subgroup 3, 201 cells/microL < CD4+ T cell counts < or = 350 cells/microL. Expression of peripheral blood Th17 and Treg cells, and number of CD4+ T cell counts were detected using flow cytometry (FCM)in HIV/AIDS patients at the pre-treatment baseline, week 4, 12, and 24, as well as those in the healthy control group.

RESULTSCompared with the healthy control group, CD4+ T cell counts and the baseline expression level of Th17 cells in the peripheral blood of HIV/AIDS patients significantly decreased, the expression level of Treg cells significantly increased P < 0.01). Compared with before treatment in the same group, CD4+ T cell counts all increased at week 4, 12, and 24 in the two treatment groups, showing statistical difference (P < 0.05, P < 0.01). There was no statistical difference in the effective rate at various CD4+ T cell levels between the two groups (P > 0.05). There was no statistical difference in expression levels of Th17 and Treg cells between the combination group and the treatment group at any time point (all P >0.05). The Th17/Treg ration significantly increased in the combination group after 24 weeks of treatment, showing statistical difference when compared with the treatment group (U = 2.135, P = 0.038).

CONCLUSIONQD could improve the immune balance of Th17/Treg cells, which might be one of its mechanisms for improving HIV/AIDS patients' immunity.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Adult ; Antiretroviral Therapy, Highly Active ; CD4 Lymphocyte Count ; Case-Control Studies ; Drugs, Chinese Herbal ; therapeutic use ; Female ; HIV Infections ; drug therapy ; immunology ; Humans ; Male ; Middle Aged ; Phytotherapy ; T-Lymphocytes, Regulatory ; cytology ; Th17 Cells ; cytology

OBJECTIVE: To investigate the proteome of hepatocyte transformation by hepatitis C virus (HCV) nonstructural protein 3 (NS3). METHODS: Human hepatocyte line QSG7701 stably expressing HCV NS3 C-terminal deleted protein was constructed, which was named pRcHCNS3/QSG. Two-dimensional electrophoresis (2-DE) was used to separate the total protein of pRcHCNS3/QSG and pRcCMV transfected cells (pRcCMV/QSG) respectively. Differentially expressed protein spots were identified by mass spectrometry. Western blot confirmed the differentially expressed proteins. RESULTS: 2-DE profiles with high resolution and reproducibility were obtained. The average spots of pRcHCNS3/QSG and pRcCMV/QSG were (1183+/-77) and (1095+/-82) respectively, and (920+/-60) spots were matched. Twenty-one differentially expressed protein spots were chosen randomly and 15 were identified by mass spectrometry. Some proteins such as Ras, P38 and HD53 which were involved in signal transduction were increased in pRcHCNS3/QSG cells. Western blot also showed strong expression of phosphorylated P44/42 and P38 in pRcHCNS3/QSG cells. Other differentially expressed proteins were related to cell cycle regulation, immunoreaction, tumor invasion and metastasis, and liver metabolizability. CONCLUSION HCV NS3 might be involved in cell malignant transformation through affecting protein expression and signal transduction such as MAPK cascade. Further study on the signal transductions and their relationship would not only be helpful to explore the mechanism of HCV related HCC, but also provide a new idea for the molecular treatment of HCC.
Cell Line ; Cell Transformation, Neoplastic ; Electrophoresis, Gel, Two-Dimensional ; methods ; Hepatocytes ; metabolism ; pathology ; Humans ; Mass Spectrometry ; methods ; Proteome ; analysis ; Proteomics ; methods ; Transfection ; Viral Nonstructural Proteins ; biosynthesis ; genetics

To study the varieties of adhesives in gels on the basis of previous studies, dosage of adhesives required for forming of gels by using the single factor design, as well as the drug loading capacity of gels with the adhesiveness and stickiness as the scoring indicators, in order to determine the forming method of analgesic micro-emulsion gel. Subsequently, the improved Franz diffusing cell method was adopted to study the release of water-soluble components and liposoluble components in analgesic gels, with imperatorin and ferulic acid as index components. The results showed that analgesic micro-emulsion gel could promote the release of imperatorin and ferulic acid. Finally, HPLC was used to detect that the loss amount of volatile components in analgesic micro-emulsion gel was 23.13% lower than that in the original analgesic gel. In the experiment, we finally prepared finished products of micro-emulsion gel, discovered that the micro-emulsion technology is helpful to improve the synchronous release of water-soluble components and liposoluble components in prescriptions and can reduce the loss of volatile components.
Adhesiveness ; Analgesics ; chemistry ; therapeutic use ; Animals ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Emulsions ; Hydrophobic and Hydrophilic Interactions ; Mice ; Ointments ; Pain ; drug therapy

Objective To develop pathogenic surveillance on measles and to effectively isolate measles virus. To know the genetic characterizations and molecular epidemiology of wildtype measles viruses from 2005 to 2007 in Guangdong Province,and provide the scientific basis for measles control and eradication. Methods Vero/ Slam cell line were used,measles viruses were isolated from throat swabs or urine specimens collected from uspected measles patients in outbreaks and sporadic patienta. A 450 nucleotides fragment of the C-terminal of the nucleoprotoin (N) gene was amplified and by RT-PCR and subjected to scquenee and phylngenetie analysis using Bio-Edit software. Results 82 wild-type measels virus were obtained from 377 throat swabs and urine specimens from 2005 to 2007 in Guangdong Province measles lab. The measles isolation rate was 23.58 % in 2005,17.11% in 2006,39.13% in 2007.The succeed rate of virus isolation is related to the quality of specimens collected and the days after rash occurrence. Conclusions We have grasped the techniealability of measles virus isolation and confirm action,and got higher isolation ratio. The wild-type measles virus isolated from Guangdong Province is of H1 genotype from 2005 to 2007,which is the same as the dominant genotype circulation.

AIMTo study the effect of modified starfish sterol [C03, succinic acid (5-epiandroene-17-one-3beta-ol) diester] on experimental arrhythmias.

METHODSArrhythmias were induced by drugs (Aco, Oua, BaCl2 and adrenalin) i.v., ligating the left anterior descending coronary artery and electricity.

RESULTSC03 71.4 mg x kg(-1) (ig) was shown to increase the dose of Oua inducing VP, VT, VF and CA in guinea pigs (P < 0.01); C03 (26.8, 80.4 mg x kg(-1)) was found to increase the dose of Aco inducing VF and CA in rats (P < 0.01); C03 (8.9, 26.8, 80.4 mg x kg(-1)) increase the dose of barium chloride and delay the onset time of ventricular arrhythmias (P < 0.01); C03 (14.1, 42.3 mg x kg(-10) shorten time of recovering induced by adrenalin in rabbits (P < 0.01); C03 (80.4 mg x kg(-1)) was shown to reduce the number of ventricular arrhythmias induced by coronary artery ligation in rats (P < 0.05), C03 increase VFT induced by electricity in rabbits, VFT of C03 14.1 mg x kg(-1) increased from (5.1 +/- 2.5) V to (11.0 +/- 2.7) V (P < 0.01), 42.3 mg x kg(-1) increased from (6.1 +/- 1.7) V to (15 +/- 5) V (P < 0.01).

CONCLUSIONStarfish sterol has anti-arrhythmic effect.

Aconitine ; Animals ; Anti-Arrhythmia Agents ; therapeutic use ; Arrhythmias, Cardiac ; chemically induced ; physiopathology ; prevention & control ; Barium Compounds ; Cats ; Chlorides ; Epinephrine ; Guinea Pigs ; Materia Medica ; isolation & purification ; therapeutic use ; Mice ; Ouabain ; Rabbits ; Rats ; Starfish ; chemistry ; Sterols ; isolation & purification ; therapeutic use ; Ventricular Fibrillation ; physiopathology

OBJECTIVETo get a more comprehensive understanding of the clinical characteristics of pediatric victims in earthquake and to summarize the experience of medical rescue.

METHODSThe clinical information was collected from the pediatric victims who were admitted to West China Hospital, Sichuan University following the Lushan earthquake in 2013 and Wenchuan earthquake in 2008. The clinical data were compared between the pediatric victims in the two earthquakes.

RESULTSThirty-four children under 14 years of age, who were injured in the Lushan earthquake, were admitted to the West China Hospital before April 30, 2013. Compared with the data in the Wenchuan earthquake, the mean age of the pediatric victims in the Lushan earthquake was significantly lower (P<0.01), and the mean time from earthquake to hospitalization was significantly shorter (P<0.01). In the Lushan earthquake, 67.6% of the injured children had variable limb fractures; traumatic brain injury was found in 29.4% of hospitalized children, versus 9.5% in the Wenchuan earthquake (P<0.05). Among the 34 children, no amputation and death occurred, and all the 13 severe cases started to recover.

CONCLUSIONSThere were higher proportions of severely injured children and children with traumatic brain injury in the Lushan earthquake than in the Wenchuan earthquake. But these cases recovered well, which was possibly due to timely on-site rescue and transfer and multi-sector, multi-institution, and multidisciplinary cooperation.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Earthquakes ; Emergency Medical Services ; Humans ; Rescue Work ; Wounds and Injuries ; epidemiology ; therapy

Objective To study the genetic characterization of enterovirus 71 (EV71) strains isolated from specimens of patients with hand-foot-mouth disease (HFMD) in Ningxia province in 2008. Methods All the stool, throat swab and vesicle samples that collected from patients with HFMD were cultured. The positive isolates were identified by reverse transcriptase PCR (RT-PCR) with specific primers of EV71. Complete VP1 gene sequences (891 nucleotides) of 29 strains (part of 93 EV71 strains) were determined and compared with A, B and C genotype reference EV71 strains while EV71 China isolates by homogeneity and phylogenetic tree analyses. Results 215 strains of EV were isolated from 439 specimens. Results from RT-PCR indicated that 93 strains belong to EV71. Phylogenetic tree analysis revealed that the selected 29 stains were clustered with reference strains of C4 subgenotype. The nucleotide identity with C4 reference strains was 91.7%-99.4%. The amino acid homogeneity was 96.6%-100.0%. Conclusion The recently identified EV71 strains in Ningxia province belonged to subgenotype C4 which resembled to most of the isolates in China.

OBJECTIVETo investigate the effectiveness of reconstruction of immunological functions of T cells on the degree of metastases of mouse hepatocarcinoma and the mechanisms of their functioning.

METHODSThe T cell model of immunological functions in Balb/c nu/nu mice was established and the effectiveness of the model was evaluated. The mice were divided into 4 groups. The immunological functions of T cells in experiment groups of Balb/c nu/nu mice were reconstructed. Metastases of the cancer in lymph nodes in each group were examined histologically. The formation time and growth rate of the tumors were calculated. The expression of MHCI and II of the tumor cell line and the difference of expression of immune associated gene were detected by Th1-Th2-Th3 gene array.

RESULTSThe ratio of CD3, CD4, CD8 and CD4/CD8 in the reconstructed group was higher than that in the control group. The average formation time was 7.7+/-0.6 days in Balb/c nu/nu mice and 11.5+/-1.3 days in Balb/c mice. The extent of metastases of the experiment group was lower than that of the control group (P < 0.05). The expression of MHCI of the high metastasis cell line was lower than that of the low metastasis cell line (P < 0.05). The expressions of Th1/Th2 associated genes in lymphocytes of high metastasis mice were lower than those of the low metastasis mice.

CONCLUSIONReconstruction of the immunological function of T cells can influence the metastasis of mouse hepatocarcinoma. The alteration of MHC molecule and low expression of Th1/Th2 correlated genes in lymphocytes may be a factor influencing the metastasis of liver cancer.

Animals ; CD4-CD8 Ratio ; Carcinoma, Hepatocellular ; immunology ; pathology ; Liver Neoplasms ; immunology ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; T-Lymphocytes ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Tumor Cells, Cultured