Contraceptive And Anti-inflammatory Effects Of Jieze No.1

Partâ… Clinical Research of JZNo.1 on Cervicitis Caused by Ureaplasma UrealyticumObjectives To observe the clinical therapeutic effect on cervicitis caused by ureaplasma urealyticumand.Methods 310 patients who suffered from cervicitis caused by ureaplasma urealyticum were randomly divided into 3 groups:Group A:the treated group integrated with traditional Chinese herbs Jieze No.1 30rains in vaginal once a day for 10days and Azithromycin orally 1.0g once 72hours for three times;Group B:the group treated with Jieze No.1 30mins in vaginal once a day for 10days;Group C:the group treated with Azithromycin orally 1.0g once 72hours for three times.Results The clinical observation showed that the overall clinical crue rate of group A reached 85.3%;group B reached 67.8%,group C reached 60.3%,The overall clinical crue rate of group A showed significant differences compared with group B and group C (P＜0.01,P＜0.01).The clearing rate of Uu in group A is 78.4%,group is 60.9%,group C is 47.9%;Group A also showed great differences in clearing rate of Uu compared with group B and group C(P＜0.01,P＜0.01).Especially to drug-resistant strain,the clearing rate of Uu in A group reached 48.1%,group B reached 42.1%,and group C reached 16.1%.The clearing rate of Uu in group A and group B has significant differences compared with group C(P＜0.01,P＜0.05).while group A and group B has no significant differences(P＞0.05).Conclusion JZ No.1 combined with azithromycin can effectively treat cervicitis caused by ureaplasma urealyticum,Eespecially it takes a remarkable effect on drug-resistant strains,It deserves further research. Partâ…¡The Effect of Jieze No.1 on Vaginal Microorganism in VitroObjectives:To observe antibacterial effect of Jieze NO.1(JZ1) on vaginal microorganism in vitro.Methods:Minimum Inhibitory Concentration(MIC) and Minimum Bactericidal Concentration(MBC) of staphylococcus aureus(SA),Escherichia coli(EC),candida albicans(CA),and streptococcu s(ST),trichomonas vaginalis and B.fragilis were detected by an agar dilution method.We then try to obtain killing curves which relate the antibacterial effect of JZ NO.1 to EC.MIC and MBC of UU were detected by a microdilution broth method.And the Kirby-Bauer method was used to measure the bacterial inhibiting diameters of Jieze NO.1 decoction on lactobacillus.CPE,MTT and plaque assay are used to estimate JZ1 antiviral activity on herpes simplex virus type 2 (HSV-2).Results:JZ NO.1 decoction produces better results on EC and SA,equivalent on ST, and not better on CA.KCS test indicate that 7.8mg·ml^-1 JZ NO.1 decoction can inhibit the growth of EC weakly,62.5mg·ml^-1 JZ NO.1 decoction can reduce the amount of EC greatly, 125 mg·ml^-1 JZ NO.1 decoction can expel EC completely.JZ NO.1 decoction has strong inhibitory effect on clinical strain Uu.After treated with Jieze NO.2 decoction of 0.25g.ml^-1 +0.0625mg.ml^-1 concentration for 2h or 0.125g.ml^-1+0.03125mg.ml^-1 concentration for 48h,the mortality of trichomonas vaginalis was all reached 100%,which was higher than that of N-9 and Jieze NO.1 decoction at the same concentration and Metronidazole of 1/100 concentration;Jieze NO.2 decoction,Jieze NO.1 decoction and Metronidazole all showed some antibacterial effect on B.fragilis,and their MIC were 0.125g.ml^-1+0.25mg.ml^-1,0.5mg.ml^-1,0.625ug.ml^-1 respectively;N-9 showed little antibacterial activation on B. fragilis,and its MIC was more than 20mg.ml^-1.50%Inhibitory Concentration(IC₅₀) of JZ NO.1 on HSV-2 is 0.89mg·ml^-1 by CPE,0.98 mg·ml^-1 by MTT.Therapeutic Index(TI) is 7.07 by CPE and 8.16 by MTT.Conclusions:The antibacterial effect of JZ NO.1 decoction on trichomonas vaginalis is better than that on EC,SA,ST,B.fragilis and CA.JZ NO.1 decoction also can inhibit the growth of Uu,and possesses antiactivity of HSV-2 in vitro, Partâ…¢The Effect of Jieze No.1 on Spermicidal Action with Nonoxynol-9 in VitroObjective:Spermicidal effect of JiezeNO.1(JZ1) decoction and Nonoxynol-9(N-9) on human sperm was examined in vitro.Methods:The minimum spermicidal concentration of JZ1 decoction,N-9 and their mixture solution in 20s and 3min were examined by improved spermicidal test of Sander-cramer in vitro.The percentage of progressively moving spermatozoa,moving spermatozoa and viable spermatozoa were also observed in 20s,3min and 30min after the liquid medicine.Results:(1) The minimal spermicidal concentration of JiezeNO.1 decoction in 3 min is 1g·ml^-1,and minimal spermicidal concentration of N-9 in 20s is 0.25mg·ml^-1.The minimal spermicidal concentration of N-9 with 1g·ml^-1 JiezeNO.1 decoction in 20s and 3 min is 0.125mg·ml^-1 and 0.0625mg·ml^-1,and it is lower than concentration of N-9 (0.25mg·ml^-1) in 20s and 3min.(2) Sperms did not recover their activitys in a revival test.when the minimum spermicidal concentration of either JZ1 decoction,or N-9,or the mixed solution of the two agents,was used.(3)N-9(JZ1 in mix group) showed significant differences in the percentages of progressively moving spermatozoa,moving spermatozoa, and visible spermatozoa in 20s,3min,and 30min,when compared with N-9 alone(P＜0.01).Conclusions:JiezeNO.1 decoction can improve N-9 spermicidal action in vitro,and also reduce the dose of N-9.when combined with N-9,it has proved of synergic effects.Partâ…£The Preventing Effect of Jieze NO.1 Gel on richomonas Vaginitis in MiceObjective To observe the preventing effect of Jieze NO.1 gel on trichomonas vaginitis in mice.Methods 112 mice were separated into Jieze NO.1 group(high titer,median titer and lower titer),Nonoxynol-9 group,Jieze NO.2 group(high titer,median titer and lower titer),Nonoxynol-9 group,Metronidazole group,gel control group and model group. Every mouse was inoculated by 20ul gel of different drugs or saline intravaginally before modeling composition,2min later,all mice were inoculated by 20ul trophozoites.We collected the lotion to test under microscope and culture in the medium at 2,4,7,10,12,14 day after inoculation,checking active trichomond and comparing infection rate between each group.Results The infection rate of Jieze NO.2 and N-9 group was 0.0%,it had significant differences(P＜0.01)compared with model group at the same time(P＜0.01);The infection rate of metronidazole group was 10.0%in 4 days,it was lower than that of control group in 10 days(P＜0.01);The infection rate of Jieze NO.1 2g.ml^-1 group was lower than that of control group before 2day(P＜0.05);The infection rate had no difference between jel control group and model group;When compared with metronidazole group,the infection rate of Jieze NO.2,N-9,2g.ml^-1 and 1g.ml^-1 of Jieze NO.1 group had no difference(P＞0.05); The infection rate of 0.5g.ml^-1 of Jieze NO.2 was lower than Jieze NO.1 at 2days(P＜0.05).Conclusion The high titer group,the median titer group and the lower titer group of Jieze NO.2 all can effectively prevent the occurrence of trichomonas vaginitis.Partâ…¤The Preventive Studies of Jieze NO.1 on Candida AlbicansObjective:To evaluate prevention effect of Jieze NO.1(JZ1) gel,the combination of JZ1 and Nonoxynol-9(JZ2)gel on Candida Albicans Vaginitis in mice.Methods:Mice were infunded intravaginally with 20ul gel of blank and drug,1min later inoculated intravaginally with 5×10⁴ stationary-phase C.albicans blastoconidia (ATCC90028) in 20μl of phosphate-buffered saline(PBS).On day 2,4,7 and 14 after inoculation,the pseudohyphae in vaginal lavage fluid,the vaginal fungal burden(CFU),the scores of inflammation in vaginal tissue,PAS of vaginal and uterus tissues were observed. And the prevention rate of vaginitis was calculated.Results:There was dosage-dependent effect of the prevention rate on JZ1 in JZ2 gel. Compared with groups of model and placebo of gel,there were significant difference on the prevention rate(65%) in group of high concentration of JZ2(P＜0.01),which could equal to the effect of group of Bifonazole Gel,and better than the group of 10%N-9(P＜0.05). There were low prevention rate(P＜0.05) in other groups of JZ2.But they were significant difference when compare with the group of model.Conclusion:JZ2 had better prevention effect on Candida Albicans Vaginitis.Partâ…¥Experimental Research on Preventing Effect of Jieze NO.1 Gel on Genital Herpes in MiceObjective:To set up genital herpes model in mice and observe the preventive effect of Jieze NO.1 gel on this model.Methods:We separated 32 mice into four groups,i.e.,the high titer group(HT),the median titer group(MT),the low titer group(LT) and saline control group(NS),All mice were injected subcutaneously with progesterone.On the fifth day,mice were anesthetized by 3%pentobarbital.The mice in virus groups are inoculated by 20ul different titers of HSV-2 invaginally,2.5×10⁶ PFU·ml^-1(HT),2.5×10⁵ PFU·ml^-1(MT),and 2.5×10⁴ PFU·ml^-1(LT),respectively;The mice in NS were given 20ul saline.Then we evaluated the success of genital herpes model in mice through genital pathology scores,viral titers in vaginal washes and ELISA for anti-HSV-2 IgG respectively.Secondly,we separated 72 mice into 6 preventive groups,i.e.,Jieze NO.1 high dose group(JZH),the median dose group(JZM),the low dose group(JZL),the ACV group(ACVR):the model group(MOD) and the saline control group(NS).At the same time of virus challenging,the mice were pretreated intravaginally with 1g·ml^-1(JZH),0.5g·ml^-1(JZM),and 0.25g·ml^-1(JZL), respectively,the mice in NS and MOD with saline intravaginally,and the mice in ACV with 12%ACV intravaginally.Genital pathology was monitored daily and scored daily,and vaginal washes were analyzed for viral titers by plaque assays.Once the experiment was finished,all mice were killed and the following steps are implemented:(1) The serum was detected for anti-HSV-2 IgG through ELISA,(2)part of the vaginal tissue was analyzed histopathologically,and(3) the other part the vaginal tissue was analyzed through FQ-PCR for HSV-2 DNA.Results:The death rate of HT in model is 50.0%,that of MT is 12.5%,while no mice in LT are dead.The pathology score of HT is higher than MT and LT.The positive rates of anti-HSV-2 IgG in both HT and MT is 100.0%and 50.0%in LT.In preventive experiment, (1) the genital pathology scores,the viral titers in vaginal washes,and HSV DNA in vaginal tissues in JZH,JZM and ACVR have significant difference in comparison to MOD(P＜0.05,P＜0.01);(2) the genital pathology scores,the viral titers in vaginal washes,and HSV DNA in vaginal tissues in both JZH and JZM are lower than those of ACVR,and have no significant difference in comparison to ACVR(P＞0.05);(3) the genital pathology scores and the viral titers in vaginal washes in JZH are higher than those JZM,although the difference are not statistically significant(P＞0.05).(4) the genital pathology scores,the viral titers in vaginal washes,and HSV DNA in vaginal tissues in JZH and JZL have significant difference in comparison to NS(P＜0.05);(5) The positive rate through ELISA of JZH,JZM,JZL,ACV and MOD is 75.0%,58.0%,41.6%and 91.6%, respectively;For comparison,the positive rate of JZH,JZM,JZL,ACV and MOD is 58.3%,41.6%,50%and 100%respectively by cell culture.(6) The mice in MOD appear dots of conjestion under their membrane,and the volume of their vaginal becomes significantly bigger.These features,however,are not observed in the mice in JZH,JZM, and JZL.(7) Lots of lymphocyte infiltrations occur and epithelium is histopathologically damaged in the mice in MOD,while few lymphocyte infiltrations in JZH,JZM,and ACV, and some lymphocyte infiltrations in JZL.Conclusions:The death rate of mice in HT is highest,but the mice in MT have low death rate and high positive rate,and the mice in LT have low positive rate,and thus,we choose the MT as the animal model for subsequent preventive experiment.JZH,JZM and ACV can relieve the vaginal pathology,reduce the viral titers in vaginal washes and HSV-2 DNA in vaginal tissues and raise the anti-HSV-2 IgG positive rate.Although JZL can not reduce the antibody positive rate and the amount of the HSV-2 DNA,but can relieve vaginal pathology,reduce the viral titers in vaginal washes slightly.Moreover,1g·ml^-1 JZ NO.1,0.5g·ml^-1 JZ NO.1 and 12%ACV can effectively prevent the occurrence of genital herpes in mice,and if genital herpes do occur,JZ1 can relieve the degree of illness.Partâ…¦Spermicidal and Intravaginal Contraception Effects of Jieze NO.1 gelObjective:To evaluate the spermicidal and contraceptive effects of Jieze NO.1 gel, nonoxynol(N-9),the combination of Jieze NO.1 and N-9(JZ2) gel in rats and rabbits.Methods:The drug was applied in female rats or rabbits intraviginally before coition, then we collected the vaginal washes in 1min,3min,5min,10min,15min and 30min to observe the percentage of moving spermatozoa,the percentage of death spermatozoa and the death time of all sperms under microscope after mating;The corpora luteum numbers and embryos were counted after 20 days,calculating the fertility rate and fertilized eggs rate.Results:The minimal spermicidal concentration of N-9 in 20s and 3min were 0.25mg/ml.The minimal spermicidal concentration of N-9 in JZ2 was 0.125mg/ml in 20s and 0.0625mg/ml in 3 min.There were not any mobile sperms in the rats vagina within 1 minute after copulation in all groups of gel of simple N-9 and JZ2.There were higher antifertility rates in groups of JZ2 gel with 10%N-9 and greater and had significant difference when compared with natural control gel and placebo gel.The antifertility rate could reach 100%in JZ2 gel with 14%N-9 and 12%N-9,equaled to the group of 14%N-9, 90%and 40%in JZ2 gel with 10%N-9 and 8%N-9,all showing significant decrease(P＜0.01),when compared with group of natural control gel(0%).The groups of JZ2 gel with 12%N-9 and 10%N-9 showed significant difference when compared with N-9 alone in the same concentration(P＜0.05) in rats.In rabbit,The percentage of moving spermatozoa and death spermatozo in group of the high,median and the low titer of Jieze NO.2 had significant difference(P＜0.01) when compared with the control group at the same time and Jieze NO.1 group in 1min,and between the low titer of Jieze NO.2 and N-9 group;The death time of all sperms in groups of the high,median and the low titer of Jieze NO.2 were 0.625±1.77min,4.00±12.25min,4.50±9.26min separately,which had significant difference(P＜0.01) when compared with the control group and between the the high titer group of Jieze NO.2 and Jieze NO.1 group,the low titer of Jieze NO.2 and N-9 group;The corpora luteum numbers had no variability between each groups;Embryos,fertility rate and fertilized eggs rate of the high titer,the median titer and the low titer of Jieze NO.2 group were lower than that of Jieze NO.1 and control group(P＜0.01),which had variability when compared between the low titer of Jieze NO.2 and N-9 group.Conclusion:JZ1 could improve N-9 spermicidal action,Jieze NO.2 gel had strong spermicidal and contraceptive effects in female rats and rabbits,of which the effect was better than that of using Jieze NO.1 gel or N-9 gel alone.Partâ…§Effect of Jieze Prescription on Lactobacillus,Glycogen and Lactoferrin in Mice VaginaObjective:To study the effect of Jieze NO.1 gel,jieze No.2 gel on Lactobacillus,glycogen and lactoferrin in mice vagina,and evaluate its side effect.Methods:180 female Kunming mice were divided into the groups of blank control, blank gel(matrix control),high(2g/ml),middle(1g / ml),low(0.5g/ml) does of JieZe NO.1 gel,high(2g/ml),middle(1g/ml),low(0.5g/ml) does of JieZe NO.2 gel and 10% Nonoxynol-9,each group observed follow indexs on 24h and 72h after injected gel intravaginally.Each mouse was subcutaneously injected physiological estradiol benzoate (E₂) 2ug,72h later infunded intravaginally with 20ul Drug gel.On 24h and 72h after injected gel intravaginally,respectively,colons-forming units(CFU) of Lactobacillus in vaginal lavage fluid from each mice was counted,glycogen content was detected by vaginal pathological section and expression of Lactoferrin in vaginal tissue by Western Blotting.Results:The results of CFU of Lactobacillus and conteat of glycogen showed there were no significant differences between groups injected drug gel intravaginally and the group of blank control, the results of groups of high,middle and low does of JieZe NO.1 gel and JieZe NO.2 gel which detected on 24h and the groups of 72h showed no significant difference too,but there were significant differences between groups injected drug gel intravaginally compared with 10%Nonoxynol-9(P＜0.01).The results of expression lactoferrin showed there were no significant differences between each two groups.Conclusion:One-time using Jieze NO.1 or Jieze NO.2 gel could have no effect on mouse vaginal lactobacillus,glycogen,lactoferrin.