Objective To investigate the inactivating effect of heat and ultraviolet(UV) light on HCV JFH-1 strain using the cell culture system. Methods The HCV JFH-1 virus stock, with an initial titer of 2.5 × 104 FFU/ml, was exposed in 56℃ water bath or to UV light for varying durations of time for explo-ring their inactivating effects on the virus. The kinetics of virus titer reduction was determined by an indirect immuno-fluorescence assay (IFA). If the cells infected with the exposed virus stock were IFA negative after three blind passages, the virus stock was considered to be inactivated completely. Results After incubation of the HCV JFH-1 virus stock (2.5 × 104 FFU/ml)in 56℃ water bath for 10 min, 20 min and 30 min, the virus titers were reduced to 1.6 × 103 FFU/ml, 3.1 × 102 FFU/ml and 3.3 × 10 FFU/ml, respectively. The exposure of the virus stock to UV light (wavelength 253.7 nm, intensity ≥60 μW/cm2, 30 cm below the UV lamp) for 15 s, 30 s and 45 s resulted in virus fiter reduction to 1.0 × 103 FFU/ml, 1.1 × 102 FFU/ml and 2.7 × 10 FFU/ml, respectively. After 40 min incubation of the virus stock at 56℃, or 1 min exposure to UV light (wavelength 253.7 nm, intensity ≥60 μW/cm2) the virus infectious titer was reduced below the detection limit of IFA, and the IFA was still negative even after three blind passages, indicating that the virus was inactivated completely. Conclusion HCV is sensitive to heat and UV light treatment. For HCV JFH-1 virus stock containing 2.5 × 104 FFU/ml virus, heat treatment at 56℃ for 40 min, or UV light expo-sure at an intensity of ≥60 μW/cm2 for 1 min, resulting in complete virus inactivation.

Antigens ; analysis ; Autoantigens ; analysis ; Breast Neoplasms ; metabolism ; Citrates ; Female ; Formaldehyde ; Hot Temperature ; Humans ; Hydrogen-Ion Concentration ; Immunohistochemistry ; methods ; Iodide Peroxidase ; analysis ; Iron-Binding Proteins ; analysis ; Paraffin Embedding ; Receptors, Progesterone ; analysis ; Thyroid Gland ; immunology ; Tissue Fixation

Antigens ; analysis ; Cervical Intraepithelial Neoplasia ; metabolism ; Citric Acid ; Cyclin-Dependent Kinase Inhibitor p21 ; analysis ; Edetic Acid ; Formaldehyde ; Hot Temperature ; Humans ; Hydrogen-Ion Concentration ; Immunohistochemistry ; Intestinal Mucosa ; immunology ; Ki-67 Antigen ; analysis ; Microwaves ; Palatine Tonsil ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; analysis

Objective To evaluate left ventricular(LV) systolic synchrony and sequence in patients with premature ventricular complexes(PVCs) from right ventricular outflow tract (RVOT).Methods Thirty patients with frequent isolated PVCs from RVOT and 30 healthy subjects as control were included.Speckle tracking imaging (STI) was performed to assess the time-to-peak segmental systolic strain in longit udinal(TsL), circumferential (TsC) and radial (TsR) direction.The standard deviation (SD) of TsL,TsC and TsR of 18 LV segments were calculated respectively.All values of patients with PVCs were recorded during sinus beats(PVC-S) and PVC beats(PVC-V) respectively.LV systolic sequence in PVC-V was analyzed.Results Significant differences were observed in the SD values between the PVC-V and control subjects in three directions,as well as between the PVC-S and control subjects in circumferential and radial direction.In PVC-V significance difference was seen in TsL and TsR from apical to basal level,as well as in TsL and TsC in different walls.Conclusions LV systolic synchrony was demonstrated in patients with PVCs from RVOT during both sinus beats and PVC beats.Systolic sequence in PVC beats from RVOT exhibit certain rules.

Objective To compare and evaluate two type-specific primers PCR genotyping methods of hepatitis B virus ( HBV) which were established by Naito et al ( Naito method) and our lab (new method). Methods The two genotyping methods were applied for detecting the plasmids containing the HBV genomes of genotype A or D or subgenotype B1 or C2 and the plasmids mixed with different proportion of subgenotypes B1 and C2. In addition, the genotypes of 113 serum samples of patients with chronic HBV infection from Shenzhen, Handan and Urumqi cities of China were identified by the two methods, respectively. The results were verified by PCR product based sequencing. Results The sensitivity of the two methods showed no difference when they were applied to detect the plasmids containing the HBV genomes of genotype A or D or subgenotype B1 or C2. While detecting the plasmids mixed with different proportion of subgenotypes B1 and C2, the sensitivity of the new method was superior than that of Naito method. Meanwhile, the specificity of the new method was obviously superior than that of Naito method. Both of the two methods were highly sensitive in identification of HBV genotypes of serum samples with a single genotype. However, the new method showed more sensitive in identification of the B/C mix strains than that of Naito method. The total coincidence rate between the two methods was 83. 2% (94/113), with the discrepancy of 16. 8% (19/113). Fifteen of the 19 inconsistent genotypes by the two methods were selected and their PCR products were sequenced directly. The sequencing results were identical with that of the new methods, but not with that of the Naito method. Conclusion The new method is more sensitive, and its specificity is superior to the Naito method. It could be used for clinical and epidemiological studies on HBV genotype and subgenotype in China.

From the hospital information system (HIS) of 20 national grade III-A general hospitals, 2 960 cases of herpes zoster as the research object, analyzes the relations between the general information, syndrome of traditional Chinese medicine (TCM), western medicine combined diseases, the relationship between the solar term and the incidence of herpes zoster, and the combined use of Chinese and western medicine. Among the patients with 46-65 year old has the highest percentage of diseased; admission to general outpatient clinic is the most; the most common medical payment is medicare; combined disease such as hypertension, diabetes and coronary heart disease is more common; early treatment effect of herpes zoster is better than the sequelae; summer and autumn solar term patients is hospitalized more, TCM syndrome is damp heat of liver fire; about drugs, western medicine is the most commonly used vitamin B1 and mecobalamin, traditional Chinese medicine is the most frequently used Danhong injection, combination therapy with promoting blood circulation drugs and neurotrophic drugs. Thus, herpes zoster, more common in elderly patients, with no obvious relationship between solar term, should be early diagnosis and early treatment, often with combination of Chinese traditional and western medicine treatment.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Herpes Zoster ; diagnosis ; drug therapy ; epidemiology ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Thiamine ; therapeutic use ; Vitamin B 12 ; analogs & derivatives ; therapeutic use ; Young Adult

OBJECTIVETo observe the effect of bear bile powder (BBP) on the STAT3 pathway and its downstream target genes of nude mice hepatocellular carcinoma (HCC) xenograft, and to explore its mechanism for treating HCC.

METHODSThe subcutaneous xenograft model was established using HepG2 cells. When the subcutaneous transplanted tumor was formed, naked mice were randomly divided into two groups, the BBP group and the control group. Mice in the BBP group were administered with BBP by gastrogavage, once daily for 3 consecutive weeks, while mice in the control group were administered with normal saline by gastrogavage, once daily for 3 consecutive weeks. The body weight and the tumor volume were measured once per week. By the end of medication, the tumor weight was weighed and the tumor inhibition ratio calculated. The apoptosis of the tumor tissue was detected by TdT-mediated dUTP nick end labeling (TUNEL). The expression of Bcl2-associated X protein (Bax), B cell lymphoma/eukemina-2 (Bcl-2), cyclin-dependent protein kinase (CDK4), cyclinD1 were detected by reverse transcription-polymerase chain reaction (RT-PCR). The protein expression levels of signal transducers and transcription activators 3 (p-STAT3), proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, CDK4, and cyclinD1 were determined by immunohistochemistry.

RESULTSBBP could inhibit the tumor volume and tumor weight, showing statistical difference when compared with the control group (P < 0.01). Results of TUNEL showed that BBP could significantly induce the apoptosis of hepatoma carcinoma cells. Results of RT-PCR showed that BBP could up-regulate the expression of Bax and down-regulate mRNA expression of Bcl-2, CDK4, and cyclinD1. Immunohistochemical results showed that BBP could up-regulate the expression of Bax and inhibit the protein expression of p-STAT3, PCNA, Bcl-2, CDK4, and cyclinD1.

CONCLUSIONBBP could induce the apoptosis of hepatoma carcinoma cells and inhibit their proliferation by regulating STAT3 pathway.

Animals ; Bile ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Ursidae ; Xenograft Model Antitumor Assays ; bcl-2-Associated X Protein ; metabolism

Objective To establish a hepatitis B virus (HBV) nested PCR (nPCR) for detection of genotypes A-D and subgenotypes B1,B2, C1 and C2. Methods The entire HBV nucleotide sequences of genotypes A-H retrieved from GenBank were compared and analyzed by DNAStar software. The PCR primers were designed by Primer Premier 5.0 software,and the nPCR for genotyping HBV/A-D as well as subgenotyping B1, B2,C1 and C2 were established. There were 3 steps in the process:step 1 for genotypes B, D and subgenotypes C1, C2 with the amplification of Mix A; step 2 for genotype A with the amplification of Mix B; step 3 for subgenotypes B1 and B2 with the amplification of Mix C in the second-ound PCR, based on the first-round amplification procedure. A total of 68 serum samples from patients with chronic HBV infection were detected by nPCR. 15 of 68 sera were selected randomly and their PCR products were directly sequenced to confirm the accuracy of the method. Results Among 68 serum samples of patients with chronic HBV infection detected by the nPCR, 23.53% (16/68) were infected with B2, 11.76% (8/68) with C1,48.53% (33/68) with C2,1.47% (1/68) with D,11.76% (8/68) with B2C2 mix strains,1.47% (1/68) with C2D mix strains and 1.47% (1/68) with B2/C1/D mix strains. The sequencing analysis of the 15 serum samples had the same results as detected by nPCR. Conclusion nPCR is a simple,rapid method and able to detect genotypes A-D and subgenotypes B1 ,B2 ,C1 and C2 subtypes of HBV with both high sensitivity and specificity.

Objective To evaluate left ventricular(LV)myocardial contraction patterns and function when pacing in different right ventricular(RV)sites and discuss echocardiogarphic method to evaluate physiologcal pacing mode.Methods This study included 26 patients with paroxysmal supraventricular tachycardia without organic heart disease.Four pacing modes including right atrium pacing(AAI),RV apex pacing(VVI-RVA),RV septal pacing(VVI-IVS)and RV outflow tract pacing(VVI-RVOT)were performed on the patients in a random order after succussful radiofrequency ablation.The parameters measured in each pacing mode included(1)LV systolic function parameters:LV twist angle(Twist),aortic systolic velocity-time integral(VTIAo)and LV global strain(Gε)；(2)LV contracting pattern:segmental peak systolic strain(Sε),the time to peak value(TPε),and the distribution of segmental Sε,TPε in each layer or wall.The relationship between Sε,TPε of each wall was analyzed.[Results]Pacing from RV sites showed lower Twist,VTIAO and Gε than AAI mode.Gε demonstrated significant difference in three RV sites pacing mode(VVI-RVOT＞VVI-IVS＞VVI-RVA,P＜0.05).Compared with the AAI mode,the distribution of segmental Sε,TPε in the each layer or wall alerted significantly in three RV sites pacing mode,especially in VV1-RVA.The distribution pattern was similar in VVI-RVOT and VVI-IVS.Furthermore,the wall Sε collated negtively with wall TPε(r =-0.51,P＜0.001).[Conclusions]Compared with AAI mode,RV pacing,especially the VVI-RVA induced the alternation of LV contraction patterns and reduction of systolic function.Longitudinal strain parameters can be used to assess the myocardial contraction patterns and function in different pacing mode.

OBJECTIVETo compare the differences in the complete genome sequence between mumps epidemic strain and mumps vaccine strain S79 isolated in Zhejiang province.

METHODSA total of 4 mumps epidemic strains, which were separated from Zhejiang province during 2005 to 2010, named as ZJ05-1, ZJ06-3, ZJ08-1 and ZJ10-1 were selected in the study. The complete genome sequences were amplified using RT-PCR. The genetic differences between vaccine strain S79 and other genotype strains were compared; while the genetic-distance was calculated and the evolution was analyzed.

RESULTSThe biggest difference between the 4 epidemic strains and the vaccine strain S79 was found on the membrane associated protein gene; whose average nucleotide differential number was 42.5 +/- 3.0 and the average variant ratio was 13.6%; while the mean amino acid differential number was 12.8 +/- 1.5 and the average variant ratio was 22.4%. The smallest difference among the 4 epidemic strains and the vaccine strain was found in stromatin genes, whose average nucleotide differential number was 73.8 +/- 2.5 and the average variant ratio was 5.9%; while the mean amino acid differential number was 3.0 +/- 0.8 and the average variant ratio was 0.8%. The dn/ds value of the stromatin genes of the 4 epidemic strains reached the highest, as 0.6526; but without any positive pressure (dn/ds < 1, chi2 = 0.87, P > 0.05). There were mutations happened on the known antigen epitope, as 8th amino acid of membrane associated protein genes and on the 336th and 356th amino acid of hemagglutinin/neuraminidase proteins. Compared with the vaccine strain, the glycosylation sites of ZJ05-1, ZJ06-3, ZJ08-1 and ZJ10-1 increased 1, 1, 2 and 2 respectively. The complete amino acid sequence of all strains showed that there were 17 characteristic sites found on the genotype-F mumps strain. Within the complete genome, the genetic-distance between epidemic strains and vaccine strains in Zhejiang province (0.071) was significantly larger than the genetic-distance between strains in Yunnan province (0.013); the difference showing statistical significance (t = 4.14, P < 0.05). Except nucleocapsid protein genes, all the genes shared similar evolution tree.

CONCLUSIONThere were significant differences found in the genes between mumps epidemic strain and mumps vaccine in Zhejiang province.

Amino Acid Sequence ; China ; epidemiology ; Genome, Viral ; Genotype ; Humans ; Molecular Sequence Data ; Mumps ; epidemiology ; genetics ; virology ; Mumps Vaccine ; Mumps virus ; classification ; genetics ; isolation & purification ; Viral Proteins ; genetics