The Clinical And Experimental Research Of The Treatment Which Activating Blood, Dissolving Stasis, Eliminating Mass And Killing Embryo On Tubal Pregnancy

IntroductionIn the recent years, with the increase of abortion and infections in the department of gynecology and obstetrics, the rate of ectopic pregnancy is rising. According to the statistics, the incidence of ectopic pregnancies among the all is above 1%[1], and the overwhelming majority of them are tubal pregnancy(95%).The etiopathogenesis of tubal pregnancy is considered to be syndrome of blood stasis in the lower abdomen by the traditional Chinese medicine. So the Chinese medicine which activating blood, dissolving stasis, eliminating mass and killing embryo has been used to treat tubal pregnancy for a long time. The treatment program is put into practice to treat the patients of tubal pregnancy who are fit the indication. The treatment program has been considered to be useful which is composed by Ectopic Pregnancy I Recipe (Salvia miltiorrhiza Bunge, Paeonia. veitchii Lynch, Peach kernel) plus Radices trichosanthis, Lithospermum erythrorhizon, Chinese formulated products for oral use, the powder of Chinese medicine for external use and the Chinese medicine parenteral solution for intervenous drop infusion. The purpose is to decrease the passions and postoperative complications because of the operations, and to assure the safety of reproductive activity.1. The clinical researchObjective: Adopt prospective disparation case-control study method to observe the clinical curative effects of the traditional Chinese medicine for treating tubal pregnancy. To evaluate the correctness and the feasibility of the "Chinese and western treatment program of tubal pregnancy"。Through the results of the statistics to prove the traditional Chinese medicine is an effective treatment to treat tubal pregnancy, and to promote and provide better clinical evidence for the further verification of rationality and utility of the Chinese and western treatment program for tubal pregnancy.Research Contents and Research MethodsThe patients of tubal pregnancy who were treated in the First Affiliated Hospital of Guangzhou University of Chinese Medicine during 2009.1-2010.12 were collected. By adopting prospective disparation case-control study method (2:1)and single blind method, the patients those who were considered to be in the same condition were divided into observative group and control group randomly, after had been distinguished the disease stages, syndrome differentiation and calculated the total score of illness impact factors. The observative group were treated by "Chinese and western treatment program of tubal pregnancy", while the control group were treated by pure western medicine-Mifepristone. The effective rate, the change trend and rules of serumβ-HCG, the treatment course of the valid cases of the two groups were collected severally in order to investigate the clinical rationality of the Chinese and western treatment program of tubal pregnancy.Results(1)The cases for test:There were 184 cases from January 2009 to December 2010, who were fit to be treated by expectant treatment by drug.119 cases were fit to be treated by pure Chinese medicine treatment among the all, which were divided into two groups, the observative group 82 cases while the control group 37 cases.65 cases were fit to be treated by conbined treatment of traditional Chinese medicine and western medicine among the all, which were divided into two groups, the observative group 45 cases while the control group 20 cases.(2)Illness impact factors:Among the 184 cases, age 25 to 29 years took up the largest proportion, reach maximum 37.5%. The onset time were 43-56 days from the last menstrual time took up the largest proportion, reach maximum 52.72%. The syndrome of vague pain took up the largest proportion, reach maximum 49.46%. The diameter<3cm of the mass through B-ultrasound examination took up the largest proportion, reach maximum 73.37%. The diameter<3cm of the internal hemorrhage through B-ultrasound examination took up the largest proportion, reach maximum 76.63%. The serumβ-HCG<1000 IU/L took up the largest proportion, reach maximum 71.2%. The total score of illness impact factors were 6.96±1.13 and 7.19±1.15 of those fit to be treated by pure Chinese medicine. The score of illness impact factors were 8.49±0.97 and 8.40±1.05 of those fit to be treated by conbined treatment of traditional Chinese medicine and western medicine.Using T-test, we can know that the there is no difference between observative group and control group in the illness impact factors(P>0.05), the two groups were comparable.(3)The results of the clinical curative effects:There is difference between observative group and control group in effective rate (P<.0.05) of the two groups which were fit to be treated by pure Chinese medicine. There is no difference between observative group and control group in effective rate (P >0.05) of the two groups which were fit to be treated by conbined treatment of traditional Chinese medicine and western medicine.There is no difference between observative groups and control groups in treatment course of different therapeutic method (P>0.05).There is difference between the two groups which were fit to be treated by pure Chinese medicine, in the time that the serumβ-HCG dropping down to 10%(P<0.05). While there is no difference between the same two groups in the time that the serumβ-HCG dropping down to 50%(P>0.05).There is no difference between the two groups which were fit to be treated by conbined treatment of traditional Chinese medicine and western medicine, in the time that the serumβ-HCG dropping down to 50% and to 10%(P>0.05).2. The experimental researchObjectiveIn the experimental research, the apoptosis and the apoptosis related proteins of the trophocytes in the fallopian tube were checked to observe the functional mechanism of the traditional Chinese medicine (Ectopic I, Decoction), which activating blood, dissolving stasis, eliminating mass and killing embryo for treating tubal pregnancy. Such study has positive influence in enriching the theory of Chinese gynecology, setting basis for clinical dialectics medication, developing the traditional Chinese medicine and providing the scientific theoretical basis for treating tubal pregnancy.Research Contents and Research Methods(1)The trophocytes cultured in vitro:The villus in the fallopian tube of those whom were fit to be treated by drugs but chose to take operation were collected. The trophocytes were cultured in vitro to be treated as research carrier.(2) The preparation of medicated serum①The medicinal broth of traditional Chinese medicine which activating blood, dissolving stasis, eliminating mass and killing embryo:The traditional Chinese medicine was thickened to be 1.5g/ml and conserved in-20℃.②The preparation of medicated serumThe SD rats were divided into five groups, three of them were feeded by traditional Chinese medicine with different dose- high dose group, moderate dose group and low dose group. The western medicine- Methotrexate had been intramuscular injected 1 hour before the serum was collected to prepare another group. The rats of the last group were feeded by normal saline.1 hour after the last time that given medicine, the serum of the SD rats were collected. The serum of the same group was mixed together, inactivated, degermed and conserved in-20℃.③Same indexThe observation of general state of health:the weights, the diet quantity and the water quantity of the rats were recorded 3 times every week. The mental state, the motoricity, the defecate situation of the rats were observed every day.The weights of the generative organs:The weights of the uteruses and ovaries of the rats were weighted in the day when they were killed.(3) Three groups of medicated serum were respectively added into cultured trophocytes cell in vitro. Then the cells were incubated for 72 hours. ①The apoptotic rate of different groups were checked by flow cytometry.②The expression of Bcl-2 and Bax were checked by the western blot technique.Results(1)The trophcyotes were cultured in vitro①Cell morphology:Most of the cells extended to be round, the number of the cells raised after 5 to 6 days. Some of them connected together and presented to be long spindle. The cell morphology presented to be epithelioid cell.②mmune fluorescence staining:The CK18 signals existed in the cells while the vimentin signals didn't.③The secretory function of the trophocytes:The expression ofβ-HCG can be checked in the culture solution of passage cell.④The growth curve of the cells:The exponential phase of growth of the trophocytes was 6 days, the Platform period of the trophocytes was 12 days.(2)The impact of the traditional Chinese medicine to ratsThere are no difference among the groups of traditional Chinese medicine and the group of the normal saline in the same time in the weights- the first day/the forth day/the seventh day after had been feeded the traditional Chinese medicine (P>0.05). There are no difference among the groups of traditional Chinese medicine and the group of the normal saline in the same time in the water quantity- the first day/the forth day/the seventh day-after had been feeded the traditional Chinese medicine (P>0.05).In the seventh day after had been feeded the traditional Chinese medicine, the diet quantity of the high dose group descended, there is significant differences between such group and the group of the normal saline(P<0.05)There are no difference among the groups of traditional Chinese medicine, the group of the normal saline and the group of western medicine in the weights of the uteruses and ovaries (P>0.05).(3)The impact of the apoptosis after the trophocytes were cultured by medicated serum that contained Chinese medicine ①After had been treated with different serum that contained different medicine in vitro for 72 hours, the apoptotic rate of the trophocytes raises, the group of Chinese medicine raised most obviously. There is difference between the group of normal saline and group of Methotrexate (P<0.05). There is significant difference between the group of normal saline and group of Chinese medicine (P<0.01). There is no difference between the group of Methotrexate and group of Chinese medicine (P>0.05)②After had been treated with different serum that contained different medicine in vitro for 72 hours, the expressions of Bcl-2 protein of the trophocytes descended, the group of Chinese medicine descended most obviously. There is difference between the group of normal saline and group of Methotrexate (P<0.05). There is significant difference between the group of normal saline and group of Chinese medicine(P0.05)③fter had been treated with different serum that contained different medicine in vitro for 72 hours, the expressions of Bax protein of the trophocytes didn't change obviously. There is no difference among the three groups (P>0.05).④he rate of Bcl-2/Bax down-regulated when treated with the serum contained Chinese medicine, and the apoptosis rate raised as a result.Research Conclusion1. After compared with the western medicine, the "Chinese and western treatment program of tubal pregnancy" was good stability in drug treatment of tubal pregnancy. Moreover, the treatment is effective and reasonable.2. Some points of the "Chinese and western treatment program of tubal pregnancy" should'be optimized through more perspective clinical research.3. The trophocytes in the fallopian tube were successfully cultivated in vitro, and proved that it can provide enough vector for further research.4. The traditional Chinese medicine that activating blood, dissolving stasis, eliminating mass and killing embryo induced the apoptosis. Inducing apoptosis may be one of the mechanisms of the effect of killing embryo.5. The traditional Chinese medicine that activating blood, dissolving stasis, eliminating mass and killing embryo deceased protein expression of Bcl-2 and deceased the rate of Bcl-2/Bax. Down-regulating of Bcl-2 may play an important role in the apoptosis in tubal pregnancy.