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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.
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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.
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The purpose is to study the prophylactic and therapeutic effect of the traditional Chinese Medicine (TCM)-Jinyebaidu (JYBD) to guinea pig cytomegalovirus (GPCMV) intrauterine infec tion. The virus-free female and male guinea pigs were screened with nest-polymerase chain reaction (N-PCR). After inbred, pregnant guinea pigs were selected and divided into 3 groups randomly: 5guniea pigs of the blank control group were not given either GPCMV or JYBD. 31 guniea pigs of the positive control group were inoculated 1 mL (107 TCID50 ) suspension of GPCMV intraperitoneal. 10 guniea pigs of the experimental group were inoculated GPCMV firstly and then perfused stomach with JYBD for 14 days (Dosage in accordance with the modulus of the weight ratio of human to guniea pig). The effects of JYBD on the intrauterine infection of GPCMV were observed.The results showed that JYBD could decrease the maternal infection rate from 100 % (31/31) to 50% (5/10) (P＜0. 001), the intrauterine infection rate from 100 % (72/72) to 75 % (21/28) (P＜0. 001), and the rate of abnormal outcome of pregnancy from 64.4 % (29/45) to 25.0 % (7/28)(P＜0. 001), the infective symptoms being relieved. It can be concluded that traditional Chinese medicine JYBD can prevent and treat GPCMV intrauterine infection, and can be expected a prophy lactic drug for HCMV intrauterine infection.
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The purpose is to study the prophylactic and therapeutic effect of the traditional Chinese Medicine (TCM)-Jinyebaidu (JYBD) to guinea pig cytomegalovirus (GPCMV) intrauterine infection. The virus-free female and male guinea pigs were screened with nest-polymerase chain reaction (N-PCR). After inbred, pregnant guinea pigs were selected and divided into 3 groups randomly: 5 guniea pigs of the blank control group were not given either GPCMV or JYBD. 31 guniea pigs of the positive control group were inoculated 1 mL (10(7) TCID50) suspension of GPCMV intraperitoneal. 10 guniea pigs of the experimental group were inoculated GPCMV firstly and then perfused stomach with JYBD for 14 days (Dosage in accordance with the modulus of the weight ratio of human to guniea pig). The effects of JYBD on the intrauterine infection of GPCMV were observed. The results showed that JYBD could decrease the maternal infection rate from 100% (31/31) to 50% (5/10) (P < 0.001), the intrauterine infection rate from 100% (72/72) to 75% (21/28) (P < 0.001), and the rate of abnormal outcome of pregnancy from 64.4% (29/45) to 25.0% (7/28) (P < 0.001), the infective symptoms being relieved. It can be concluded that traditional Chinese medicine- JYBD can prevent and treat (GPCMV intrauterine infection, and can be expected a prophylactic drug for HCMV intrauterine infection.
Subject(s)
Cytomegalovirus , Cytomegalovirus Infections/drug therapy , Drugs, Chinese Herbal/therapeutic use , Fetal Diseases/drug therapy , Fetal Diseases/prevention & control , Fetal Diseases/virology , Phytotherapy , Pregnancy Complications, Infectious/drug therapy , Random AllocationABSTRACT
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.
Subject(s)
DNA, Viral/analysis , Infectious Disease Transmission, Vertical , Female Urogenital Diseases/virology , Papillomaviridae/isolation & purification , Papillomavirus Infections/transmission , Polymerase Chain Reaction/methods , Polymorphism, Restriction Fragment Length , Pregnancy Complications, Infectious/virologyABSTRACT
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.
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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.
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The aim of the study was to investigate the correlation between prognosis of human cytomegalovirus (HCMV) infection and zinc in mature women, in order to explore the effect of serum zinc on HCMV infection. 900 mature woman were examined for zinc level and serum HCMV specific antibodies (IgG and IgM). 63 HCMV-IgM positive cases were divided into 3 subgroups according to their serum zinc level, and HCMV-IgM was observed for conversion after treatment with a Chinese traditional medicine (Jinyebadu). Our results showed that the mean concentration of zinc in serum was significantly lower in HCMV-IgM(+) group than that in HCMV-IgM(-) group (P<0.001). A positive correlation was found between the response of mature females with HCMV infection to the treatment and the levels of serum zinc (P<0.001). The response of mature women with HCMV is poor when the serum zinc is lower than 0.70.
Subject(s)
Adult , Female , Humans , Antibodies, Viral , Blood , Antiviral Agents , Therapeutic Uses , Cytomegalovirus , Allergy and Immunology , Cytomegalovirus Infections , Blood , Drug Therapy , Drugs, Chinese Herbal , Therapeutic Uses , Immunoglobulin G , Blood , Immunoglobulin M , Blood , Phytotherapy , Zinc , BloodABSTRACT
The value of ELISA, N-PCR and RT-PCR in clinical practice for pregnant women with HCMV infection was investigated. 5581 pregnant women were screened by ELISA. Among them, 100 cases were positive for IgM (group 1), 69 for both IgM and serous DNA (group 2) and 69 for both IgM and mRNA (group 3). The infectious status, maternal-fetal transmission and pregnancy outcome were monitored. It was demonstrated that the accordance rate of group 3 and group 2 with group 1 was 56.25% and 43.75%, respectively. The maternal-fetal transmission rate in the group 1, 2 and 3 was 19.00%, 40.58% and 46.15%, respectively, with a significant difference found between group 2, 3 and group 1 (P < 0.01). Incidence of spontaneous abortion, fetal death, fetal abnormality and neonatal death in group 1, 2 and 3 was 10.00%, 15.94% and 30.77%, respectively, and that of group 3, 2 was 4 and 2 times as much as that of group 1, respectively (OR = 4.00, P < 0.001; OR = 2.343, P < 005, respectively). It was concluded that HCMV-IgM(+) can only be considered as an screening indicator for pregnant women with HCMV infection, while IgM(+) combined with serous DNA(+) or mRNA(+) indicates active infection and has a high incidence of maternal-fetal transmission and abnormal pregnancy outcome.
Subject(s)
Adult , Female , Humans , Pregnancy , Antibodies, Viral , Blood , Cytomegalovirus , Allergy and Immunology , Cytomegalovirus Infections , Diagnosis , Virology , DNA, Viral , Blood , Enzyme-Linked Immunosorbent Assay , Evaluation Studies as Topic , Immunoglobulin M , Blood , Infectious Disease Transmission, Vertical , Polymerase Chain Reaction , Methods , Pregnancy Complications, Infectious , Virology , Pregnancy Outcome , Prenatal Diagnosis , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To study the diagnostic value of human cytomegalovirus (HCMV) late mRNA detection in active intrauterine infection.</p><p><b>METHODS</b>The HCMV late mRNA in peripheral blood of 42 HCMV IgM positive pregnant women and their fetal attachments (such as chorionic villi, amniotic fluid, umbilical blood and placenta) were detected by reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Late mRNA was detected in 23 of 42 HCMV IgM positive cases, a rate of 54.3%. Fetal appendages in 13 cases of late mRNA positive mothers were also tested, of which 7 were positive, with a vertical transmission rate of 53.8%. In 12 late mRNA negative mothers, only 1 case of fetal appendages tested was positive, with a vertical transmission rate of 8.3%. There was significant difference between the transmission rates of these two groups.</p><p><b>CONCLUSIONS</b>Positive results of HCMV IgM cannot accurately reflect the activity of HCMV at the time of testing. However, the activity of HCMV is closely related to the mother-fetus vertical transmission rate. As an indicator of active HCMV infection, late mRNA can not only reflect the mother-fetus transmission rate during active HCMV infection, but also provide some information about HCMV activity in fetal tissue.</p>
Subject(s)
Female , Humans , Pregnancy , Antibodies, Viral , Blood , Cytomegalovirus , Genetics , Allergy and Immunology , Cytomegalovirus Infections , Diagnosis , Immunoglobulin M , Blood , Pregnancy Complications, Infectious , Diagnosis , RNA, Messenger , Blood , Reverse Transcriptase Polymerase Chain Reaction , Sensitivity and SpecificityABSTRACT
To investigate the clinical value of three-dimensional ultrasonography (3DUS) in obstetrics, various 3DUS rendering methods including surface mode, transparent mode and multiplanar mode were employed to scan 30 fetuses in second and third trimester by using the transabdominal volume transducer. The results showed that surface mode could vividly demonstrate the surface morphologic features of the fetuses, as well as the stereo-shape and the spatial relationship among the surface structures. The face, limbs, umbilical cord and outer genitalia of the fetus could be well displayed by surface mode. Transparent mode could reveal the bony structures under the surface, such as ribs, vertebrae, crania, etc. The result was not affected by the sophisticated curvature of these bony structures and the success rate was up to 100 %. When rendered by multiplanar mode, the region of interest (ROI) could be viewed from different directions. It should be concluded that 3DUS could serve as a supplement to two-dimensional ultrasonography (2DUS). 3DUS might play an important role in prenatal diagnosis and enhance the diagnostic confidence level of the physicians.
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To investigate the clinical value of three-dimensional ultrasonography (3DUS) in obstetrics, various 3DUS rendering methods including surface mode, transparent mode and multiplanar mode were employed to scan 30 fetuses in second and third trimester by using the transabdominal volume transducer. The results showed that surface mode could vividly demonstrate the surface morphologic features of the fetuses, as well as the stereo-shape and the spatial relationship among the surface structures. The face, limbs, umbilical cord and outer genitalia of the fetus could be well displayed by surface mode. Transparent mode could reveal the bony structures under the surface, such as ribs, vertebrae, crania, etc. The result was not affected by the sophisticated curvature of these bony structures and the success rate was up to 100 %. When rendered by multiplanar mode, the region of interest (ROI) could be viewed from different directions. It should be concluded that 3DUS could serve as a supplement to two-dimensional ultrasonography (2DUS). 3DUS might play an important role in prenatal diagnosis and enhance the diagnostic confidence level of the physicians.
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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.
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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.
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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
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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.
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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
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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