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.

OBJECTIVETo study the predicting effect of proly 4-hydroxylase beta polypeptide (P4HB) in treating non-small cell lung cancer (NSCLC) patients by Yiqi Chutan Recipe (YCR).

METHODSTotally 46 stage III and IV NSCLC patients were treated by YCR for 4 therapeutic courses. Effect was assessed by RECIST of solid tumor. P4HB expression was detected in the lung cancer tissue by immunohistochemical assay. Factors affecting disease control rates (DCR) of YCR were analyzed by Logistic regression analysis. The correlation between P4HB expression and the effect of YCR was analyzed.

RESULTSThe DCR of advanced NSCLC treated by YCR was 36.96% (17/46 cases). P4HB was high expressed in advanced lung cancer tissue (6/15 cases). Gender, initial treatment, and retreatment are independent factors for affecting DCR of treating lung cancer by YCR.

CONCLUSIONP4HB might be taken as a factor for predicting the effect of YCR in treating NSCLC.

Carcinoma, Non-Small-Cell Lung ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Lung ; Lung Neoplasms ; drug therapy ; metabolism ; Male ; Procollagen-Proline Dioxygenase ; metabolism ; Protein Disulfide-Isomerases ; metabolism

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.

Objective To analyze the complete genome sequence of Guangxi HEV isolate swGX32 and to compare it with other HEV isolates. Methods The overlapping fragments of HEV isolate swGX32 were amplified with reverse-transcription nested polymerase chain reaction (RT-nPCR),and the 5′ and 3′ ends of viral genome were amplified with rapid amplification of cDNA ends (RACE). The PCR products were cloned and sequenced. The sequence and phylogenetic analysis of swGX32 was performed. Results The genome of swGX32 consisted of 7240 nt excluding the polyA tail, with 4 nt overlapping between ORF1 and ORF2. ORF3 is contained in the sequence of ORF2. The complete genome sequence of swGX32 shared identity of 73%-74%, 73%, 74%-75%,83%-94% with HEV genotype 1,2,3 and 4, respectively. Among all these HEV reference sequences, swGX32 showed the highest identity with the human isolate JKO-ChiSai98C (94%). Phylogenetic tree showed that swGX32 belonged to genotype 4 and clustered with JKO-ChiSai98C in the branch of HEV subtype 4a. Conclusion The swine HEV isolate swGX32 is closely related to human strain JKO-ChiSai98C genetically and phylogenetically, which further provides molecular biology evidence of hepatitis E as a zoonosis.

Objective To investigate the efficacy and toxicity of neoadjuvant regional arterial chemotherapy in the treatment of advanced gastric cancer. Methods The clinical data of 158 patients with advanced gastric cancer and with the same clinical stages who were admitted to Renji Hospital of Shanghai Jiaotong University from February 2002 to May 2005 were retrospectively analysed. Preoperative regional arterial chemotherapy was applied to 76 patients (test group) and the remaining 82 patients only received surgical treatment (control group). The chemotherapy regimen was epirubicin (50 mg/m2) + cisplatin (60 mg/m2) + 5-fluorouracil (1000 mg/m2).This regimen was modified to oxaliplatin (130 mg/m2) + 5-fluorouracil (1000 mg/m2) since 2003, and surgery was performed 6-11 days after the chemotherapy. All patients received postoperative intravenous chemotherapy.The clinical effects, radical resection rate, operative complications and long-term survival of the two treatment methods were evaluated. All data were analysed using the chi-square test and Kaplan-Meier analysis. Results The radical resection rate was significantly higher at 86% (65/76) in the test group compared with 71% (58/82)in the control group ( x2 = 5.01, P ＜ 0. 05 ). The toxicity of the chemotherapy in the test group was mild. The postoperative complication rate was 20% (15/76) in the test group and 16% (13/82) in the control group, with no significant difference between the two groups (x2 = 0.41, P＞0.05). The median survival time was 41 months in the test group and 23 months in the control group. The 5-year overall survival rate was higher in the test group (44.6%) than that in the control group (29.1%) (x2 =3.95, P＜0. 05). Conclusions Neoadjuvant regional arterial chemotherapy is well tolerated by patients with advanced gastric cancer. It is also effective for increasing the radical resection rate and improving the long-term survival.

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

Animals ; Hepatitis E ; prevention & control ; Hepatitis E virus ; immunology ; Humans ; Viral Hepatitis Vaccines

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

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.

Animals ; Genotype ; Hepatitis E ; veterinary ; virology ; Hepatitis E virus ; genetics ; Hepatitis, Viral, Animal ; virology ; Prevalence