FAILURE OF HERPES SIMPLEX VIRUS TYPE 2 TO SUBSTITUTE FOR DIMETHYL-BENZANTHRACENE IN TWO-STAGE SKIN CARCINOGENESIS

There are some Indications thtt herpes aimplex virus (HSV) may be mutagenic. Specific chromosomal changes have also been demonstrated In cultured cells infected with HSV. To further Investigate the mutagenic activity of HSV type 2 (HSV-2) we used mouse skin as a model system for cardnogenetls. Inoculation of the back skin of 4-week-old Sencar mke with live virus twice per week for one week or with Inactivated virus twice per week for two weeks was used to Initiate the mouse akin. After Initiation with HSV-2, 12-O-tetradecanoylphorbol-13-acetale(TPA) was applied twice weekly for 50 weeks as a promoter. During a period of 52 weeks, no skin carcinoma was found In the experimental groups, whereas 55% of control mice treated with 9, 10-dlmethy1-1, 2- benzanthracene (DMBA ) and then with TPA-developed skin carcinoma. The results demonstrate that HSV-2 could not substitute for DMBA in this animal model of two-stage skin carcinogenesis.

SEARCH FOR HERPES SIMPLEX VIRUS TYPE2（HSV－2）AND HUMAN PAPILLOMAVIRUS（HPV）IN THE NORMAL AND ABNORMAL CERVICAL SAMPLES

The specimens of 111 cervical carcinomas. 68 chronic cervicitis and 43 normal cervical exfoliated epithelial cells were examined for the presence of HSV2 DNA sequences with DNA hybridization using HSV2 BgL Ⅱ N fragment probe labelled by 32PdCTP. The result showed that the infection rates of HSV2 in the samples of cervical cancer.chronic cervicitis and normal epithelial cells were 1 4. 41 %(16/111). 27.94%( 19/68) and 25.58% ( 11/43),respectively. It was implied that early stages carcinogenesis of cervical epithelial cells might be correlated with the HSV2 infection.Sixteen HSV 2 positive samples of cervical carcinomas were also examined for the presence of the sequences homologous to human papillomavirus (HPV) type 6B/11. 16 and 18 DNA using dot blot hybridization (Tm17℃). The result indicated that 13 out of 16 were HPV 16 DNA hybridization positive accounting for 81. 2% of all HSV-2 positive samples and none of them were positive for HPV type 6B/11 and 18. The result indicated that double infection of HSV 2 and HPV16 in the same cervical carcinoma tissues may provide a strong evidence of the viral synergistic interaction in the induction of female cervical

ENHANCEMENT OF MORPHOLOGICAL AND ONCOGENIC TRANSFORMATION OF MOUSE CELLS WITH 12-O-TETRADECANOYL PHORBOL- 13-ACETATE FOLLOWING HERPES SINPLEX VIRUS TYPE 2 INFECTION

Sequential exposure of mouse embryo cells to HSV-2 and TPA gave rise to a synergistic enhancement of the transformation frequency. The transformants were selected for their ability to form dense foci of cells in medium containing 10% or 1%(low) fetal bovine serum. The average number of foci induced with HSV -2 followed by TPA was about 3 or 5(in low serum) fold greater than that induced with HSV- 2 alone. HSV- 2 antigen could be detected in about 10% of transformed cells before 27th passage with immunofluorescence technique. Of two cell lines established from single focus , one designated BL which was preferable to form foci in subcultures was tumorigenic after 21th passage. All of the tumors were sarcomas with interlacing bundles of pleomorphic fibroblasts. The other,designated NP was nontumorigenic until 50th passage. The BL cell line was composed of two distict cell types, i. e.,pigmented and unpigmented. No viral DNA sequences weredetected in the cells of tumors derived from BL cell line.

The Chinese herbal prescription JieZe-1 inhibits caspase-1-dependent pyroptosis induced by herpes simplex virus-2 infection in vitro

Objective:JieZe-1(JZ-1),a Chinese herbal prescription,has an obvious effect on genital herpes,which is mainly caused by herpes simplex virus type 2(HSV-2).Our study aimed to address whether HSV-2 induces pyroptosis of VK2/E6E7 cells and to investigate the anti-HSV-2 activity of JZ-1 and the effect of JZ-1 on caspase-1-dependent pyroptosis.Methods:HSV-2-infected VK2/E6E7 cells and culture supernate were harvested at different time points after the infection.Cells were co-treated with HSV-2 and penciclovir(0.078125 mg/mL)or caspase-1 inhibitor VX-765(24 h pretreatment with 100μmol/L)or JZ-1(0.078125-50 mg/mL).Cell counting kit-8 assay and viral load analysis were used to evaluate the antiviral activity of JZ-1.Inflammasome activation and pyroptosis of VK2/E6E7 cells were analyzed using microscopy,Hoechst 33342/propidium iodide staining,lactate dehydrogenase release assay,gene and protein expression,coimmunoprecipitation,immunofluorescence,and enzyme-linked immunosorbent assay.Results:HSV-2 induced pyroptosis of VK2/E6E7 cells,with the most significant increase observed 24 h after the infection.JZ-1 effectively inhibited HSV-2(the 50%inhibitory concentration=1.709 mg/mL),with the 6.25 mg/mL dose showing the highest efficacy(95.76%).JZ-1(6.25 mg/mL)suppressed pyroptosis of VK2/E6E7 cells.It downregulated the inflammasome activation and pyroptosis via inhibiting the expression of nucleotide-binding oligomerization domain-like receptor family pyrin domaincontaining protein 3(P<0.001)and interferon-γ-inducible protein 16(P<0.001),and their interactions with apoptosis-associated speck-like protein containing a caspase recruitment domain,and reducing cleaved caspase-1 p20(P<0.01),gasdermin D-N(P<0.01),interleukin(IL)-1β(P<0.001),and IL-18 levels(P<0.001).Conclusion:JZ-1 exerts an excellent anti-HSV-2 effect in VK2/E6E7 cells,and it inhibits caspase-1-dependent pyroptosis induced by HSV-2 infection.These data enrich our understanding of the pathologic basis of HSV-2 infection and provide experimental evidence for the anti-HSV-2 activity of JZ-1.

THE CERVIX MULTI－VIRUSES INFECTION AND THE DEVELOPMENT OF CERVICAL CARCINOMAS

The infections of human papilloma virus (HPV),herpes simplex virus type 2 (HSV-2) and Epstein-Barr virus (EBV) to human cervical epithelium are universal and some of them are associated with the development of cervical carcinomas. One hundred and eleven cervical cancer biopsies taken from Beijing area and Xiangyuan county in Shanxi province of China were examined for the presence of DNA sequences of HPV, HSV-2 and EBV by means of Dot blot hybridization. The experiment results showed that the total infectious rates of HPV, HSV-2 and EBV were 71.17% (79/111), 14.44%(16/111) and 15.63% (15/96), respectively. Seventy-nine samples positive for HPV were further analysed for the viral types distribution, the result indicated that the positive specimens of HPV type 16 accounted for 72.15%, otherwise, the biopsies positive for HPV type 18and 6B/11 only accounted for 16-46% and 11.39%,respectively. The data suggested that HPV infection,especially HPV type 16, may play an important role in the development of cervical carcinomas- 16 specimens positive for HSV-2 were examined for HPV DNAsequences and the result uncovered that 13 out of them were HPV16 positive (81.25%), 11 samples containing EBV genomes were also examined for HPV DNA sequences and the result indicated that 9 of 11 were detectable for HPV DNA. The experiment results proved a direct evtidence of multi-virus infection in cervix and of the synergistic interaction among viruses in the process of cervical epithelial carcinogenesis.Comparing of the viruses' infection of two areas, the frequencies of HPV infection in Beijing and Xiangyuan areas were 72.84% (59/ 81) and 66.67% (20/30) , the infectious rates of HSV-2 in the two areas were 8.64%(7/81) and 30% (9/30) (P<0.05), the rates of EBV infection in the two areas were 12.5% (10/80) and 31.25% (5/16)(0.1>P>0.05). The results proved another strong evidence that the high incidence of cervical cancer in Xiangyuan county may be closely correlated with multiviruses infection and with multi-virus synergistic interaction.

Disseminated herpes simplex infection induced hepatitis during pregnancy mimicking HELLP syndrome,a diagnostic challenge

The disseminated herpes simplex virus 2(HSV-2)carries a high mortality rate in pregnant women if left unrec-ognized.It often presents as unrelieved fever and hepatitis.Diagnosis is challenging due to vague symptoms and potential overlap with other conditions.Pregnancy is a risk factor as it conforms to a partially immunocompro-mised state.Dissemination to the brain could be devastating,and the treatment requires intravenous antivirals like acyclovir.Fetal outcomes are variable based on previous case reports.We present a case of young female gravida 1 para 1 who presented with disseminated HSV infection mimicking HELLP(Hemolysis,Elevated Liver enzymes,and Low Platelets)syndrome.She responded well to intravenous acyclovir,and the fetus had a viable outcome at the 26th week of gestation.Early diagnosis can prevent progression to fulminant liver failure and the need for a liver transplant.

THE INFECTION OF EBV FOR CERVICAL EPITHELIUM-A NEW CAUSITIVE AGENT IN THE DEVELOPMENT OF CERVICAL CARCINOMAS?

Epstein-Barr virus was considered as a caustive agent for Burrkitt' s lymphoma and non-malignant B lymphocytes proliferation. The recent studies revealed the striking association of the Infection of EBV with the development of human epithelial tumors. 43 specimens of normal exfoliated cervical epithelial cells, 47 biopsies of chronic cervlcitis and 80 tissue samples of cervical carcinomas were tested for the presences of EBV W fragments by using dot blot hybridization method. The results showed that the detectable rates of EBV DNA sequences In the normal exfoliated epithelium, the chronic cervlcitis and cervical carcinomas were 44.16%, 12.77% and 13.75%, respectively. Eleven EBV positive DNA samples from cervical cancers were also examined for the presence of HPV DNA. The result showed 9 out of 11 were HPV DNA positive, the cultanious infectious rate of both viruses was about 81.81%.In this paper, the EBV genomes existed In the part of biopsies of cervical carcinomas were first reported. The results Implied that the development of cervical cancer might be associated with asymptomatic EBV Infection In early stage and might be the result from the synergistic carcinogenesis of EBV with other cervical oncovinises, especially with HPV.