OBJECTIVES: To explore the mechanism through which moxibustion promotes endometrial repair in rats with in thin endometrium (TE). METHODS: Female SD rats were randomized into control group, 95% anhydrous ethanol-induced TE model group and moxibustion (at "Guan Yuan") group. High-throughput sequencing was used to identify the target genes of TE, and the targeting relationship between miR-223-3p and NLRP3 was verified using a dual luciferase assay. Histopathological of rat uterus was observed with HE staining, and expressions of miR-223-3p and NLRP3 were detected using RT-qPCR; serum levels of IL-1β and IL-18 of the rats were detected using ELISA, and protein expressions of NLRP3, ASC, caspase-1 and GSDMD in the uterus were detected with Western blotting. The pregnancies of the rats after treatment were counted. RESULTS: Enrichment analysis of the differential genes suggested up-regulated inflammatory response in TE, and dual luciferase assay verified targeted inhibition of NLRP3 expression by miR-223-3p. The rat models of TE had significantly decreased endometrial thickness and reduced endometrial glands and blood vessels with enhanced mRNA expression of NLRP3, increased serum levels of IL-1β and IL-18, up-regulated protein expressions of NLRP3, ASC, caspase-1 and GSDMD, lowered pregnancy rates on both the affected and unaffected sides and the overall number of pregnancies. Treatment of the rat models with mo-xibustion obviously increased the endometrial thickness and the density of glands and blood vessels, up-regulated miR-223-3p expression, lowered serum IL-1β and IL-18 levels and the protein expressions of NLRP3, ASC, caspase-1 and GSDMD, and significantly increased the number of pregnancies. CONCLUSIONS Moxibustion at "Guan Yuan" acupoint up-regulates the expression of miR-223-3p, which results in targeted inhibition of NLRP3 to suppress pyroptosis and promote endometrial repair in rat models of TE.
Animals ; Female ; MicroRNAs/genetics* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Endometrium/pathology* ; Rats, Sprague-Dawley ; Rats ; Moxibustion ; Pyroptosis ; Up-Regulation ; Interleukin-1beta/metabolism* ; Interleukin-18 ; Caspase 1/metabolism*

Objective To compare the reparative effects of moxibustion at"Guanyuan"and"Shenshu"points in rats with a thin endometrium.Methods Thirty-two female Sprague-Dawley rats were divided randomly into control(CON),model(MOD),Guanyuan(GY),and Shenshu(SS)groups.A thin endometrium model was established using 95％anhydrous alcohol.The Guanyuan and Shenshu groups underwent moxibustion at the Guanyuan and Shenshu points,respectively,and the other two groups of rats were fixed in the same way.Temperature changes at the acupoint area were measured at different time points.The pathological morphology of the uterus was observed by hematoxylin and eosin(HE)staining,and serum levels of inflammatory factors and hormone levels were detected by enzyme-linked immunosorbent assay(ELISA).Expression levels of proliferation and endometrial receptivity factors were detected by immunohistochemistry(IHC),and angiogenesis factors were detected by Western blot.Results The temperatures immediately after moxibustion and after 5 min were higher at the Shenshu compared with the Guanyuan acupoint(P＜0.01).Endometrial thickness and numbers of glands and blood vessels were decreased in the MOD group compared with the CON group(P＜0.01),serum interleukin 1 β(IL-1 β)was increased(P＜0.01)and PROG was decreased(P＜0.01),and Ki67,proliferating cell nuclear antigen(PCNA),leukemia inhibitory factor(LIF),integrin β3(ITG-β3),angiopoietin-1(ANG-1),and CD34 protein in uterine tissue were all decreased(P＜0.01).Endometrial thickness and numbers of glands and blood vessels were increased in the GY and SS groups compared with the MOD group(P＜0.01),while serum IL-1 β was decreased(P＜0.05),E2 and PROG were increased(P＜0.05),and serum interleukin 10(IL-10)was increased in the SS group(P＜0.05).Ki67,PCNA,LIF,and ITG-β3 were all increased in both treatment groups compared with the MOD group(P＜0.01).PCNA was higher in the GY group compared with the SS group(P＜0.05),while LIF and ITG-β3 were higher in the SS group compared with the GY group(P＜0.01,P＜0.05,respectively).CD34 and VEGFA levels were increased in the two treatment groups compared with the model group(P＜0.01).ANG-1 was higher in the GY group than that in the SS group(P＜0.01),while CD34 was higher in the SS group than that in the GY group(P＜0.05).Conclusions Moxibustion at the Guanyuan and Shenshu points could help to repair the pathological thin endometrium in rats.The GY group may be superior to the SS group for reducing the levels of serum IL-1 β,increasing E2,and promoting the expression of proliferation factors,while the SS group may be superior to the GY group for increasing the levels of PROG and promoting the expression of endometrium receptor-related factors.Both acupoints can promote the expression of angiogenesis factors,potentially acting via different pathways.

Objective To compare the reparative effects of moxibustion at"Guanyuan"and"Shenshu"points in rats with a thin endometrium.Methods Thirty-two female Sprague-Dawley rats were divided randomly into control(CON),model(MOD),Guanyuan(GY),and Shenshu(SS)groups.A thin endometrium model was established using 95％anhydrous alcohol.The Guanyuan and Shenshu groups underwent moxibustion at the Guanyuan and Shenshu points,respectively,and the other two groups of rats were fixed in the same way.Temperature changes at the acupoint area were measured at different time points.The pathological morphology of the uterus was observed by hematoxylin and eosin(HE)staining,and serum levels of inflammatory factors and hormone levels were detected by enzyme-linked immunosorbent assay(ELISA).Expression levels of proliferation and endometrial receptivity factors were detected by immunohistochemistry(IHC),and angiogenesis factors were detected by Western blot.Results The temperatures immediately after moxibustion and after 5 min were higher at the Shenshu compared with the Guanyuan acupoint(P＜0.01).Endometrial thickness and numbers of glands and blood vessels were decreased in the MOD group compared with the CON group(P＜0.01),serum interleukin 1 β(IL-1 β)was increased(P＜0.01)and PROG was decreased(P＜0.01),and Ki67,proliferating cell nuclear antigen(PCNA),leukemia inhibitory factor(LIF),integrin β3(ITG-β3),angiopoietin-1(ANG-1),and CD34 protein in uterine tissue were all decreased(P＜0.01).Endometrial thickness and numbers of glands and blood vessels were increased in the GY and SS groups compared with the MOD group(P＜0.01),while serum IL-1 β was decreased(P＜0.05),E2 and PROG were increased(P＜0.05),and serum interleukin 10(IL-10)was increased in the SS group(P＜0.05).Ki67,PCNA,LIF,and ITG-β3 were all increased in both treatment groups compared with the MOD group(P＜0.01).PCNA was higher in the GY group compared with the SS group(P＜0.05),while LIF and ITG-β3 were higher in the SS group compared with the GY group(P＜0.01,P＜0.05,respectively).CD34 and VEGFA levels were increased in the two treatment groups compared with the model group(P＜0.01).ANG-1 was higher in the GY group than that in the SS group(P＜0.01),while CD34 was higher in the SS group than that in the GY group(P＜0.05).Conclusions Moxibustion at the Guanyuan and Shenshu points could help to repair the pathological thin endometrium in rats.The GY group may be superior to the SS group for reducing the levels of serum IL-1 β,increasing E2,and promoting the expression of proliferation factors,while the SS group may be superior to the GY group for increasing the levels of PROG and promoting the expression of endometrium receptor-related factors.Both acupoints can promote the expression of angiogenesis factors,potentially acting via different pathways.

Objective:To construct and validate a predictive model for the risk of excessive blood loss in pa-tients with ruptured tubal pregnancy,and to provide a basis and tool for the assessment of changes in the condi-tion of patients with ruptured tubal pregnancy.Methods:Clinical data of inpatients with ruptured tubal pregnancy from January 2014 to July 2021 were retrospectively analyzed,who underwent surgical treatment in the Depart-ment of Gynecology,Dongguan Maternal and Child Health Hospital.The pelvic blood volume was categorized into excessive blood loss and non-excessive blood loss groups based on whether the amount of pelvic blood was found to be≥750 ml intraoperatively.Factors influencing the occurrence of excessive blood loss were screened and modeled by univariate analysis,Lasso regression,and multi-factor Logistic stepwise regression.The area un-der the subject working characteristic curve(AUC)was used to evaluate the discrimination of the predictive mod-el,the model's consistency was evaluated by calibration curve and goodness-of-fit test,and the clinical utility of the model was evaluated and validated by the decision analysis curve.Finally,column line plots were drawn.Results:①A total of 386 patients with ruptured tubal pregnancy were included,of whom 124(32.12%)had blood loss≥750 ml.②The optimal predictors for predicting concomitant blood loss in patients with ruptured tubal preg-nancy were screened,including:days of abdominal pain,dizziness,pallor,fatigue,the maximum diameter of para-metrial mass,human chorionic gonadotropin(β-hCG),and hemoglobin(Hb)and the model and the column line graphswere constructed accordingly.③The prediction model AUC was 0.827(95%CI 0.781-0.873);the cut-off value was 0.391,at which point the specificity and sensitivity were 68.55%and 84.35%,respectively,and the AUC validated within the model by resampling was 0.804.Clinical decision curves showed that the threshold probability intervals for the maximum net benefit values ranged from 8.5%-97%,respectively.Conclusions:The constructed prediction model was validated to suggest good discriminatory efficacy and degree of consistency.As a tool,it has clinical application value in predicting the risk of hemorrhage in patients with ruptured tubal pregnan-cy.It can help to determine the occurrence of adverse events such as hemorrhagic shock at an early stage and improve the success rate of rescue treatment.

Objective:To compare the difference on the positive detection rate of the main aeroallergens and food allergens specific IgE measured with the systems between fluorescence enzyme immunoassay(FEIA, referred to as ImmunoCAP system)and western blot(referred to as Allergy Screen system), in order to provide a basis for the rational application of methods and interpretation of the test results.Methods:The clinical information and sIgE test results data were collected from a total of 458 cases of allergic diseases from October 2017 to April 2019 in the outpatient clinic of Allergy Department in Beijing Children′s Hospital.All of the 458 cases were detected for a panel of common aeroallergens and food allergens sIgE level in their serum with the Allergy Screen system.Simultaneously, the above 141 cases were detected main aeroallergen and food allergen sIgE with ImmunoCAP system, while 303 cases only for aeroallergens and 14 cases only for food allergens.All of the cases were divided into three different phenotype groups according to the main target organ and diagnosis such as airway allergic disease group( N=293), skin allergic disease group( N=14)and multi-system allergic disease group( N=151). Meanwhile, three different age groups were referred to as <3 years old group( N=97), 3 to 6 years old group(3 years and 6 years included)( N=186)and >6 years old group( N=175). The same kinds of the allergens included in the two systems were house dust mite(HDM), cat dander, dog dander, egg white, milk, crab, shrimp, therefore data of sIgE to those seven allergens were compared. Results:In all the enrolled cases, the positive detection rate of HDM, egg white and milk sIgE detected by ImmunoCAP system were significantly higher than those by Allergy Screen system(30.6%, 36.1% and 43.2% vs 21.2%, 21.3% and 21.3%). Among the disease groups, the positive detection rate of HDM sIgE detected by ImmunoCAP system was significantly higher in the airway allergic disease group and multi-system allergic disease group than those by Allergy Screen system, respectively(33.7% and 24.6% vs 23.4% and 16.2%). The positive detection rate of egg white and milk sIgE detected by ImmunoCAP system in the multi-system allergic disease group was significantly higher than those by Allergy Screen system, respectively(47.6% and 47.6% vs 26.2% and 25.2%). There was no significant difference in the positive detection rate of cat dander, dog dander, crab and shrimp sIgE among the disease groups.The positive detection rate of HDM sIgE detected by ImmunoCAP system in all of the age groups were significantly higher than those by Allergy Screen system, respectively(14.9%, 26.9% and 42.3% vs 5.7%, 17.6% and 32.6%). The positive detection rate of cat dander sIgE detected by ImmunoCAP system in <3 years old group was significantly lower than that by Allergy Screen system(6.9% vs 16.1%). The positive detection rate of egg white sIgE detected by ImmunoCAP system in the <3 years old group and the 3 to 6 years old group were higher than those by Allergy Screen system, respectively(47.1% and 33.3% vs 32.4% and 15.0%). The difference in the positive detection rate of milk sIgE detected by these two methods in the <3 years old group and the 3 to 6 years old group was similar to the egg white.Conclusion:According to the analysis of seven kinds of major aeroallergen and food allergen sIgE results, there are differences between the two methods to detect HDM, cat dander, egg white and milk sIgE.It depends on the type of allergic disease and age.The positive detection rates of dog dander, crab, and shrimp sIgE detected by the two methods are consistent.In clinical application, comprehensive analysis should be made, testing methods should be selected rationally, and the results should be interpreted scientifically.

BACKGROUND: Thin endometrial diseases are a challenge in clinical treatment at present. Scholars have found that bone marrow mesenchymal stem cells (BMSCs) transplantation has its unique curative effect and advantages, but few studies have been conducted on pathway or gene control. OBJECTIVE: To observe the effect of BMSCs transplantation in rats with thin endometrium based on the HOXA10 regulatory network. METHODS: Twenty-one adult female Sprague-Dawley rats of SPF grade (provided by the Animal Experimental Center, Guangzhou University of Chinese Medicine in China) were randomly divided into three groups (n=7/group): control group, model group, and BMSCs group. In the latter two groups, a thin endometrium model was prepared in each rat by filling the uterine cavity with 95％ ethanol. In the control group, normal saline was injected to fill the uterine cavity of rats. After extraction of ethanol or normal saline, the rats in the BMSCs group were injected intrauterinely with 1 mL of BMSCs suspension (1×1010 cells/L) , and those in the control and model groups were given the same volume of normal saline. After two estrous cycles, the uterus of each rat was removed. Hematoxylin-eosin staining was used to measure the thickness of the endometrium. Immunohistochemistry was used to detect the expression of vimentin, keratin, vascular endothelial growth factor, leukemia inhibitory factor and integrin αvβ3. qRT-PCR was used to detect the relative transcription of HOXA10 and miR-196 b. RESULTS AND CONCLUSION: (1) Compared with the control group, the endometrial thickness of the rats were significantly thinner in the model and BMSCs groups (P < 0.05) , while the endometrial thickness in the BMSCs group was thicker than that in the model group (P < 0.05). (2) The mean absorbance values of endometrial vimentin, keratin, vascular endothelial growth factor, leukemia inhibitory factor and integrin αvβ3 were highest in the control group, higher in the BMSCs group and lowest in the model group, and there were significant differences between groups (P < 0.05). (3) The relative transcript level of HOXA10 gene in the model and BMSCs group was significantly lower than that in the control group, while the relative transcript level of HOXA10 gene in the BMSCs group was significantly higher than that in the model group (P < 0.05). The relative transcript level of miR-196 b in the model and BMSCs groups was significantly higher than that in the control group (P < 0.05) , while the relative transcript level of miR-196 b in the BMSCs group was lower than that in the model group (P < 0.05). (4) HOXA10 was negatively correlated with miR-196 b gene, HOXA10 was positively correlated with the protein expression to different extents, and miR-196 b gene was negatively correlated with the protein expression to different extents. These findings suggest that BMSCs transplantation can improve the endometrial thickness and relevant protein levels of thin endometrium rats to some extent, which may be attributed to the negative regulation of HOXA10 gene by miR-196 b, and HOXA10 gene further promotes the expression of vascular endothelial growth factor, leukemia inhibitory factor and integrin αvβ3 proteins.

Objective To establish a rat xenograft model of human uterine leiomyoma using immunosuppressive a-gent and provide a useful tool for the study on uterine leiomyoma. Methods Intragastric administration with immunosup-pressive agent mycophenolate mofetil(MMF)(40 mg/kg)was given to rats for two weeks before the surgery until the end of the experiment. 20 SPF female Wistar rats were randomly divided into 4 groups after abdominal transplantation of human leiomyoma tissues:group A received femoston containing 0.4 mg/kg estradiol and 2 mg/kg dydrogesterone, group B re-ceived estadiol 0.4 mg/kg,group C received dydrogesterone 2 mg/kg,and group D served as the control group, received distilled water 1 mL/200 g. All rat received the corresponding drugs once per day for 2 days. Samples were taken at 4 weeks after the surgery to observe the pathology of the tumor tissues. Results The modeling success rates were 90% in the group A,40 % in the group B,and 0% in the groups C and D. Conclusions Rat xenograft model of human leiomyoma can be successfully established using an immunosuppressive agent femostone with a high modeling success rate and low cost. It can be used as a new animal model for the study of transplanted leiomyoma.

Objective To observe the changes in estrous cycle and vaginal smears in ovarectomized NOD/SCID mice.Methods To continuously observe the estrous cycle time by vaginal smears of NOD/SCID mice in consecutive nine days, twice daily.After ovariectomy, the changes of estrous cycle were observed by vaginal smears for 7 days.Results The estrous cycle in NOD/SCID mice was 4-6 days.Regular estrous mice accounted for 80%.There was no significant correlation between vaginal opening and estrous cycle status.After ovariectomy, the vaginal smears showed characteristics of metestrus or diestrus.Conclusions Vaginal smear cytology can be used to determine the estrous cycle and characteris-tics of NOD/SCID female mice.The ovariectomized operation of NOD/SCID female mice is effective.

Thin endometrium is an important factor influencing the in vitro fertilization and embryo transfer, and there is a lack of effective therapy in the treatment of thin endometrium.The aim of this study was to explore a stable animal model of effective thin endometrium, and to promote the research on thin endometrium pathogenesis and provide experimen-tal basis of treatment.To analyze a variety of establishments of endometrial damage animal model reported in the domestic and foreign literatures,it is concluded that perfusing 95% ethanol into uterine cavities of rats can establish a rat model of thin endometrium,and put forward some experience and methods for its improvement.

Objective To investigate the efficacy comparison of endovascular therapy and simple medical therapy for symptomatic intracranial artery stenosis. Methods A total of 145 patients with intracranial artery stenosis were analyzed retrospectively. They were divided into either an endovascular therapy group (n=72) or a medical therapy group (n=73). They were treated with endovascular therapy (gateway balloon,wingspan stents,Apollo stents) or medical therapy (aspirin 100 mg/d,clopidogrel 75mg/d, and atorvastatin 20-40 mg/d) according the willingness of the patients or their family members. The incidences of stroke and transient ischemic attack ( TIA ) , and restenosis rate ( stenosis rate >50% as a standard) during 1-,3-,6-,9-,and 12-month follow-up periods were observed and compared. Results On the basis of medical therapy,the patients of the endovascular therapy group were successfully stented. The success rate of stenting was 98. 6% (70/71). Seven patients had complications in the endovascular therapy group (9.9%),2 of them complicated with hemorrhage(one of was died),drinking cough,hoarseness, dizziness,headache,and excitement were one case in each, the other patients were cured and discharged with active medical treatment, and they did not have serious sequelae. At 12 months after treatment, the stroke recurrence rate of the endovascular therapy group was 8. 4% (n=6,both were TIA),and that of the medical therapy group was 26. 0% (84. 2% was minor stroke). There was significant difference (χ2 =7. 752,P<0. 01);at 12 months after treatment,the incidences of restenosis and aggravated stenosis were 5. 6% (n=4) and 6. 8% (n=5) respectively. There was no significant difference (χ2 =0. 091,P>0. 05). Conclusion Compared with the medical therapy,the efficacy of endovascular therapy for symptomatic intra-cranial arterial stenosis is more significant. The improvement of clinical prognosis is superior to medical therapy.