| Genital herpes, is a common sexually transmitted diseases (STD) caused by type Ⅱ herpes simplex virus (HSV-2) infection,which is the main reason of the STD clinic genital ulcers. HSV-2can infect the host through various ways, of which the mucous membranes of the genitourinary tract is most susceptible.Mainly behave as the genitalia set of small blisters or pustules and cervical ulcers Chronic cervicitis is a common gynecological diseases, cervical erosion is the main clinical manifestations and its causes are complex. Recent studies show that a variety of sexually transmitted diseases were closely related to the incidence of cervicitis and cervical cancer.among this Patients who were infected with herpes simplex virus Type2have recurrent features and the relationship between HSV infection and the cervical erosion has gradually attracted the attention of the people.Due to the paticular structure of the cervix,HSV infection in cervix can not easily be diagnosed.The recurrent infection of HSV-2lead to cervical local congestion or ulcers, its also easily combined with other infections, which accelerated partial pathological damage and acute cervicitis easily converted into a chronic cervicitis, while the degree of cervical erosion is increasing. With repeated stimulation of HSV-2infection, the virus can make cervical epithelial dysplasia and squamous metaplasia, and even cancer.Many scholars have detected HSV-2antigen, virus fragments and particles in cervical cancer cells. The detection rate of Genital herpes with cervical erosion are increased, and the detection rate of HSV-2infection in cervical secretions was reach to32.9%in abroad Therefore, in clinical, the detection of herpes simplex virus can not be ignored to the patients with recurrent cervical erosion.Virus culture and PCR are the two main methods to monitor HSV excretion in Clinical, between this two methods,viral culture is the gold standard.but it has the disadvantage of istedious operation Jong cycle, technically demanding, Difficult to genotyping, and the test results are easily influenced by transit time and transit processing, which would finally lead to limitations in routine clinical applications. Compared with virus culture,PCR method has the advantage of high sensitivity.specificity, short time taking, and can greatly improve diagnostic accuracy, which could provide an important basis for early clinical diagnosis and treatment,and now had used gradually in clinical.Due to the special expression form of HSV-2infection, the treatment of HSV-2has always been difficulties in clinical and the treatment in clinical is the Research direction.The current multi-drug therapy.such as acyclovir, valaciclovir can be used to shorten the course and reduce relapses, although the anti-viral drugs in the treatment of the herpes simplex virus infection has little drug resistance reports to the general population, but for immunosuppressed patients, antiviral drug resistance has reacht to14%.With the development of medical technology, organ transplant patients increased.transplant recipients need to use drugs to suppress rejection,the immune system of this patients are in an inhibitory state, making HSV-2attack host more easily. The Clinical symptoms present frequently recurring and involving multiple internal organs, which had caused new problems on therapy. Select effective, safe and repeatable method of treatment to control the disease, to relieve patients’pain is a clinical problem ought to be solved.Photodynamic therapy (PDT) involves the use of a photosensitizing agent, which may require metabolic synthesis, followed by light activation. PDT requires the simultaneous presence of a chemical photosensitizer, appropriate wavelength light and oxygen inside the affected tissue. The photosensitizer is accumulated in target cells and adsorbs energy directly from a light source, which it then transfers to molecular oxygen to create highly atomic group and reactive oxygen species. These cytotoxic agents react with cellular components to cause damage that ultimatelyleads to pathological cell death to treat diseases.PDT is the treatment manner of microinvasion, no-thermogenesis and causing histoclasia in local.accompanning further studies of novel photosensitizer and the mechanism of PDT, PDT is being studied extensively for its possible application in the treatment of non-oncological diseases,such as nevus flammeus, actinic keratosis,acne and psoriasis for the past few years.Several studies have found that PDT exerts a good cytocidal activity against some viruses.The mechanism of PDT to kill the virus is that porphyrin photosensitizers had a affinity to the viral envelope lipid. The role of photodynamic therapy on the enveloped viruses is superior to the non-enveloped viruses. Since the envelope of the virus comes from the host plasma membrane.virus-infected cell membrane and viral envelopeare all such photosensitizer binding site. After Photosensitization.the viral envelope integrity is destroyed. Herpes simplex virus is an enveloped virus.therefore PDT had direct inactivation of herpes simplex virus. After ALA-PDT, stand-early gene of the virus ICP027, late gene VP16and gD were dropped by an average, so some experts have speculated that PDT inhibit proliferation of the virus through interfere viral replication necessary gene synthesis due to less accumulation of photosensitizer in normal tissue, so PDT does not cause Photosensitization of normal tissue cells. Research has shown that that photodynamic therapy had inactivation on HSV-1in vitro.Objectives The incidence of genital herpes gradually increased, and the early diagnosis and treatment to HSV-2infection can helps to effectively control the spread of the disease. The study was starting from the detection and treatment of HSV-2infection.to supply a laboratory reference for clinical diagnosis and treatment of genital herpes.1.A real-time quantitative PCR method was applied to detect HSV-2infection in patients with cervical erosion, to explore the proportion of herpes simplex virus (HSV-2) in cervical specimens from patients with cervicitis and to assess the relationship between cervical erosion and HSV-2infection.2.ALA drug and laser was used to processed human keratinocytes cell Hacat strains infected with HSV-2in vitro,to study the kill efficiency of ALA photodynamic therapy on herpes simplex virus, in order to explore the feasibility of ALA-PDT in the treatment of genital herpes and to provide a preliminary laboratory basis for the treatment of genital herpes.Medthods1.Real-time PCR was used to detect HSV-2for the262patients who were cervicitis and vaginal secretions anormal but without herpes genitalis case history from clinic service of dermatology and department of gynaecology and obstetrics.the control group were50normal female by health examination.the relation of the two groups in hsv-2infection and the level of cervical erosion were compared.2.Hacat cells in vitro culture were administered different density of5-ALA ranged from Ommol/l to4nmol/l.after3hrs incubation at37℃and5%CO2, the cells were exposure to laser irradiation doses0J/cm2to21J/cm2.MTT colorimetric method was used to detect cell survival ratio. The effect of different density and energy on keratocyte was observed..3.(1)PDT treat on the exracellular viruses:Hacat cells were implanted in mixed liquor combined HSV-2with0.5mmol/l ALA after3h adsorption.and laser irradiation doses used from6,9,12,15J/cm2.(2)PDT do with the viruses of CPE.pathological.hacat cells were implanted in HSV-2,administered different doses of laser radiation from6to15/cm2after cpe appeared. After2d of two different treatments,OD value was detected by MTT and the cytoprotection ratio was calculation. we investigated the kill effect of the5-ALA-PDT to HSV-2.Results1. QT-PCR was used to detect HSV-2:cases were found positive(30.9%)of the262patients with cervicitis and8(9.2%) of the79controls were positive for HSV-2, there were statistically significant differences in detection rate between cervicitis group and control group (χ2=16.21, P<0.05).2.The relationship of HSV-2and different degree of cervical erosion:The infection rate of HSV were117.0%(15/88),27.9%(24/86) and47.7%(42/88)。 respectily with light,modera and severe cervicitis degree (Z=-4.395, p<0.001).The infection rate of HSV-2increased with the increment of cervicitis degree(rs=0.272,P<0.050).3.The use of photodynamic therapy on HaCat, when choose different drug concentration and different laser energy:The survival rate becoming less and less with increasing concentrations of drugs and the difference has significance between the different concentration groups (F=1178.32, P<0.001).Also there were Significant difference between the different energy group (F=1001.56, P<0.001) and the survival rate getting lower and lower with increasing energy.There was interaction effect between the concentration group and the energy group (F=60.5. P<0.001).4. The role of photodynamic therapy on herpes simplex virus,(1) The killing effect of5-ALA-PDT on extracellular HSV-2:the survival rate of HaCat was statistically significant difference between the groups (F=79.38, p<0.001);when the concentration of5-ALA-PDT in the0.5mmol/1, with the increase of energy density, cell viability was gradually increasing.There were statistically significant difference among cell control group and Virus control group,0.5mmol/1.6J/cm2,0.5mmol/1.9J/cm2(p<0.001); There were statistically significant difference among virus control group and the cell control group or0.5mmol/1.9J/cm2,0.5mmol/1.12J/cm2,0.5mmol/1.15J/cm2(p<0.001);(2) The killing effect of5-ALA-PDT on intracellular HSV-Ⅱ:the survival was statistically significant difference between the groups (F=17.01p<0.001); with the increase of energy density, survival rate of HaCat was gradually increasing.There were statistically significant difference among cell control group and virus control group,0.5mmol/1.6J/cm2,0.5mmol/1.9J/cm2(p <0.001); There were statistically significant difference among virus control group and the cell control group or0.5mmol/1.12J/cm2,0.5mmol/1.15J/cm2(p<0.001).Conclusion1. The infection rate of HSV-2were higher in patinents with cervical erosion than normal contral group.The infection rate of HSV-2increased with the increament of cervicitis degree, HSV-2infection is related to cervicitis.2.5-ALA-PDT showed drictly killing effect on HSV-2in extracellar.protect HaCat cells against infection.; for those cell had cytopathic changed,5-ALA-PDT inhibit viral relication within the cell,prevent the vius spread to the surrounding HaCat cells. |
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