AIM:To study the therapeutic effect of traditional Chinese medicine Reduqing on human cytomegalovirus (HCMV). METHODS:The expression of the HCMV late-mRNA in infected cells was measured by quantitative reverse transcription-polymerase chain reaction(RT-PCR) and the development of cytopathic effect (CPE) caused by HCMV was observed before and after treatment with Reduqing. On the basis of its anti-HCMV activity study in vitro, Reduqing was applied to the child-bearing aged women and pregnant women who were HCMV-IgM positive which indicated the active HCMV infection. The serum levels of tumor necrosis factor-alpha (TNF-α) and interleukin-2 (IL-2) before and after treatment were measured by the ELISA method. RESULTS:The results showed that the Reduqing had a strong inhibitory effect on both HCMV-mRNA and CPE. The level of the TNF-α increased while IL-2 decreased significantly in HCMV-IgM positive serum when compared with HCMV-IgM negative cases. In HCMV-IgM positive patients, after treatment with Reduqing, the serum levels of TNF-α and IL-2 returned to normal, and 73 % of them turned to be HCMV-IgM negative, which was significantly higher than that (14 %) in control group. CONCLUSION:It is suggested that serum TNF-α and IL-2 level are closely related to the activity of HCMV. Reduqing can enhance the immunity of the body and has an obvious inhibition on HCMV in vitro and in vivo, and can thus serve as an effective medicine for active HCMV infection therapy, especially for the pregnant women.

AIM: To investigate whether human cytomegalovirus(HCMV) regulate human embryonic lung fibroblast(HEL) cell proliferation and apoptosis by activating NF-?B. METHODS: Immunohistochemistry and Western blot analysis were used to detect the NF-?B translocation and/or Bcl-2 and the levels of I-?B? during HCMV infection. Apoptotic cell were examined by flow cytometry, and the HEL cell proliferation was determined by MTT. RESULTS: The levels of NF-?B in the nucleus reached highly 48 h postinfection, and the levels of I-?B? were low 24 h postinfection. The activity of NF-?B was inhibited 120 h postinfection. The levels of bcl-2 was accorded with the activity of NF-?B. HCMV promoted HEL cells to proliferate before 72 h postinfection and induced apoptosis 120 h postinfection. CONCLUSION: NF-?B plays a role in HEL cell proliferation and apoptosis during HCMV infection, and it involves in the pathological mechanisms of diseases associated with HCMV infection.

AIM: To explore the relationship between human cytomegalovirus(HCMV) gene activity and cytopathogenesis in HCMV infection. METHODS: HCMV echelin-infected cell model was set up in vitro by coincubating passage cultured HEL and HCMV with different titers (group A: 10-5; group B: 10-3; and group C: 10-1). FQ-PCR was performed to evaluate the number of HCMV DNA copies. MCP mRNA was measured by RT-PCR. Meanwhile the development of cytopathologic effects (CPE) was observed under microscope and ultrastructural changes determined by TEM.RESULTS: Compared with group B and C in high HCMV titer, group A with low viral titer showed low HCMV DNA load (P

Objective To evaluate the testing value of ELISA, N-PCR and RT-PCR in clinical practice for pregnant women with HCMV infection. Methods 5581 pregnant women were screened by ELISA. 100 cases with positive IgM (group 1), 69 positive IgM combined with positive serous DNA (group 2) and 69 with positive mRNA (group 3) were studied on maternal-fetal transmission and pregnancy outcome. Results The accordance rate of group 3 and group 2 with group 1 are 56.25% and 43.75%, respectively. The maternal-fetal transmission rate in three groups is 19.00%,40.58% and 46.15%, respectively. A significant difference existed between group 2, 3 and group 1(P0.05). The total rate of spontaneous abortion, fetal death, fetal abnormality and neonatal death are 10.00%,15.94% and 30.77% in different groups, respectively and that of group 3, 2 are 4 and 2 times as much as that of group 1, respectively (OR=4.00, P

Objective To explore prospectively the relationship between treatment of toxoplasmosis and pregnancy outcome. Methods Detected by ELISA and PCR,59 pregnant women and 91 women at reproductive age with infection were treated by oral spiramycin. Then their Tox-lgM and DNA negative conversing rates,intrauterine transmission rate and incidence of abnormal pregnancy outcome were studied in contrast to those of 60 women in pregnancy and 79 at childbearing age with infection but without management. Results In healing group,the seroconversion rate of Tox-lgM and Tox-DNA from positive to negative were 83.05% (49/59) in pregnant women and 70.33%(64/91) in women at childbearing age,respectively. The rates in control group were 35% (21/60) and 37.97% (30/79). The rates of intrauterine transmission and abnormal pregnancy outcomes of healing group were 8.4% (5/59) and 30% (15/50),respectively. While those of control group were 11.86% (7/59) and 40% (20/50) accordingly (? 2=11.4970,P

Objective To study the state of herpes simplex virus (HSV) infection and the vertical transmission between pregnant women and their fetus/newborns. Methods ELISA was applied to screen HSV specific antibodies in 455 pregnant women. HSV type 2 DNA was detected by PCR technique in the tissues and appendages of fetus, whose mothers had positive HSV IgM and/or type 2 IgG antibodies. Results The infective rate of HSV types1 and 2 during pregnancy were 85.27% and 47.69%, respectively. The intrauterine infective rate was 4.24%. The positive rate of HSV type 2 IgG in the pregnant women with history of abnormal pregnancy was significantly higher than that in the control group. HSV type 2 DNA was found in the amniotic fluid and tissues of placenta and fetus. Conclusion The HSV infection was common during pregnancy. HSV type 2 infection was closely related to the abnormal pregnancy, such as abort, premature delivery and malformation. ELISA combined with PCR technique is a valuable method for the diagnosis of HSV infection.

To explore the effect of [Ca2+]i and neuronal mitochondria transmembrane potentials in hippocampus of murine cytomegalovirus (MCMV) infected mice, newborn Balb/c mice were randomly divided into two groups: a virus inoculated group and a control group. After 56 days, single cell of hippocampus was isolated, and mitochondria transmembrane potentials and the intracellular free calcium level [Ca2+]i in hippocampus were measured by means of flow cytometry (FCM). Compared with the control group, the mitochondria transmembrane potentials was decreased (P<0.01) and the intracellular free calcium level [Ca2+]i was increased (P<0.01) in inoculated group. The dysfunction of [Ca2+]i and mitochondria transmembrane potentials in hippocampus may play an important role in the functional disorders in CMV-infected CNS.

To further investigate the vertical transmission route of human papillomavirus (HPV) and the indication for the choice of mode of delivery, the infective status of 152 asymptomatic pregnant wemen and the maternal-fetal transmission were studied. By using general primers in polymerase chain reaction (GP-PCR) combined with restriction fragment length polymorphism analysis, HPV DNA positive rate in cervical secretions and venous blood in asymptomatic pregnant women was 36.21% and 52.78%, respectively, and the identified genotypes were mainly HPV16 and 18. The maternal-fetal transmission rate of HPV via genital tract as well as blood was 40.91% and 57. 89%, respectively. It was concluded that besides the transmission route of genital tract and amniotic fluid, there was also transplacental transmission of HPV in utero. Therefore,in our opinion, it is not an absolut indication to perform a cesarean delivery for the pregnant women with HPV asymtomatic genital infection.
DNA, Viral/analysis ; Disease Transmission, Vertical ; Female Urogenital Diseases/*virology ; *Papillomaviridae/isolation & purification ; Papillomavirus Infections/*transmission ; Polymerase Chain Reaction/methods ; Polymorphism, Restriction Fragment Length ; Pregnancy Complications, Infectious/*virology

Objective To construct human cytomegalovirus pp65 prokaryotic expressing vector and induce specific T lymphocyte immune response with recombinant pp 65 protein of human cytomegalovirus(HCMV) to observe the effect of CTL on infected HEL cells. Methods The whole cDNA of HCMV pp65 was amplified by PCR and inserted into prokaryotic expressing vector pRSET by gene engineer technique. The product was purified by affinity chromphotography and identified with Western blot after that recombinant plasmid was expressed by the induction of IPTG. With MTT technique, we observed the stimulating and proliferating effect of recombinant HCMV pp65 protein on PBMC in vitro. Cytotoxicity of PBMC on HCMV-infected HEL cells was detected. Results The pp65 prokaryotic expressing vector was successfully constructed and could express in engineering bacteria DE3. High dose of pure recombinant protein was acquired and had been identified. The rhHCMV pp65 protein can activate PBMC and cause the proliferation of it in vitro. The proliferated PBMC have the specific cytotoxicity to HEL cells infected by HCMV. Conclusions The acquired recombinant HCMV pp65 protein could induce specific T cells immune response in vitro to kill the HCMV infected HEL cells. And it is very important for the immune therapy of the HCMV infections.

Summary: To further investigate the vertical transmission route of human papillomavirus (HPV) and the indication for the choice of mode of delivery, the infective status of 152 asymptomatic pregnant wemen and the maternal-fetal transmission were studied. By using general primers in polymerase chain reaction (GP-PCR) combined with restriction fragment length polymorphism analysis, HPV DNA positive rate in cervical secretions and venous blood in asymptomatic pregnant women was 36.21 % and 52.78 %, respectively, and the identified genotypes were mainly HPV16 and 18. The maternal-fetal transmission rate of HPV via genital tract as well as blood was 40.91 % and 57.89 %, respectively. It was concluded that besides the transmission route of genital tract and amniotic fluid, there was also transplacental transmission of HPV in utero. Therefore,in our opinion, it is not an absolut indication to perform a cesarean delivery for the pregnant women with HPV asymtomatic genital infection.