Objective:To study the effects of intervention of Jiawei Tongmai Huazhi Decoction in the Wnt/β-catenin pathway on apoptosis of lesion cells in mice with adenomyosis (AM); To discuss its mechanism of action.Methods:The AM mouse model was established using tamoxifen. The mice were divided into model group, Jiawei Tongmai Huazhi Decoction group, and progesterone group according to random number table method, with 7 mice in each group. Additionally, a blank group of 7 female mice was set up. Jiawei Tongmai Huazhi Decoction group received oral administration of Jiawei Tongmai Huazhi Decoction at a dosage of 36.51 g/kg/day, once daily. The progesterone group received oral administration of progesterone at a dose of 0.32 mg/kg twice a week. The blank group and model group received oral administration of the same volume of physiological saline once daily. After 2 months of intervention, the morphology of uterine tissues was observed by HE staining. The levels of carbohydrate antigen 125 (CA125) and prolactin (PRL) in the serum were measured by ELISA. The mRNA levels of Wnt3a and β-catenin in uterine tissues were determined by PCR. The protein expressions of Wnt3a, β-catenin, Bax, and Bcl-2 in uterine tissues were detected by Western blot.Results:Compared with the model group, the levels of serum CA125 and PRL were reduced in the Jiawei Tongmai Huazhi Decoction group ( P<0.05). The protein expressions of Wnt3a, β-catenin, and Bcl-2 were also reduced ( P<0.05), the protein expressions of Wnt3a, β-catenin, and Bcl-2 decreased ( P<0.05), while the protein expressions of Bax increased ( P<0.05). Conclusion:Jiawei Tongmai Huazhi Decoction alleviates the progression of lesions by reducing serum CA125 and PRL levels in AM model mice, and can down-regulate Bcl-2 expression and up-regulate Bax expression, promoting apoptosis of ectopic lesion cells in mice. Its mechanism of action may be related to the inhibition of Wnt/β-catenin pathway related expression proteins.

Objective:The aim of this study is to establish the reference interval of refined immune cell subsets by multi-parameter flow cytometry.Methods:In this cross sectional study, a total of 326 healthy participants were included and divided into two groups based on age: 18-40 years old group and 41-60 years old group. Peripheral venous blood was collected in a fasting status. Flow cytometry tests were performed according to previous consensus article. The analysis of reference interval was conducted according to the documents of Clinical and Laboratory Standards Institute (CLSI) EP28-A3c and Health Industry Standards of the People′s Republic of China WS/T 402-2024.Results:The T,B,NK,DC and monocyte refined immune cell subsets applicable to the reference range of the general population mainly include: CD3 +(56.4%-83.3%),CD4 +TEMRRA (0.2%-11.6%), CD4 +TEM (14.9%-52.8%), CD4 +CD28 +(76.3%-99.9%), CD19 +CD5 +(9.7%-45.8%), CD19 +CD27 -(45.7%-90.0%), CD19 +CD27 +(9.8%-54.0%), CD3 +CD16 +CD56 +(1.3%-20.2%), CD3 -CD19 -CD20 -CD14 -CD56 -HLA-DR +(0.4%-2.0%), Monocyte Mo1 subset CD14 +CD16 -(46.3%-94.6%), monocyte Mo2 subset CD14 +CD16 +(2.8%-49.7%), etc. When the Z-value between different age groups was higher than Z* (3.50 cut-off value), the reference intervals of these subsets should be established independently according to age. Conclusions:In this study, the reference intervals of refined subsets of immune cells by multi-parameter flow cytometry has been preliminarily established. For those subgroups that meet the grouping criteria, age should be fully considered in clinical applications and laboratory validation.

Objective:The aim of this study is to establish the reference interval of refined immune cell subsets by multi-parameter flow cytometry.Methods:In this cross sectional study, a total of 326 healthy participants were included and divided into two groups based on age: 18-40 years old group and 41-60 years old group. Peripheral venous blood was collected in a fasting status. Flow cytometry tests were performed according to previous consensus article. The analysis of reference interval was conducted according to the documents of Clinical and Laboratory Standards Institute (CLSI) EP28-A3c and Health Industry Standards of the People′s Republic of China WS/T 402-2024.Results:The T,B,NK,DC and monocyte refined immune cell subsets applicable to the reference range of the general population mainly include: CD3 +(56.4%-83.3%),CD4 +TEMRRA (0.2%-11.6%), CD4 +TEM (14.9%-52.8%), CD4 +CD28 +(76.3%-99.9%), CD19 +CD5 +(9.7%-45.8%), CD19 +CD27 -(45.7%-90.0%), CD19 +CD27 +(9.8%-54.0%), CD3 +CD16 +CD56 +(1.3%-20.2%), CD3 -CD19 -CD20 -CD14 -CD56 -HLA-DR +(0.4%-2.0%), Monocyte Mo1 subset CD14 +CD16 -(46.3%-94.6%), monocyte Mo2 subset CD14 +CD16 +(2.8%-49.7%), etc. When the Z-value between different age groups was higher than Z* (3.50 cut-off value), the reference intervals of these subsets should be established independently according to age. Conclusions:In this study, the reference intervals of refined subsets of immune cells by multi-parameter flow cytometry has been preliminarily established. For those subgroups that meet the grouping criteria, age should be fully considered in clinical applications and laboratory validation.

NONMEM

Objective To investigate the influence of different types of pneumoperitoneum on the ultrastructure of the extraperitoneal muscular tissues in rats. Methods By using pulse-mode or constant-pressure mode insufflators,we established CO2 pneumoperitoneum in 80 rats (40 for each) with the pressure set at 15 mmHg. At 60,90,120 and 180 minutes after the insufflation,the muscular tissues below the peritoneum of the rats (10 rats at each time point) were obtained to observe the ultrastructural change under a transmission electron microscope (TEM). Ten rats that received anesthesia only were set as a control. Results In both the groups,vacuolar degeneration was found in the myocytes below the peritoneum after 60 minutes of pneumoperitoneum,and became more and more marked with time. Moreover,the phenomenon was more significant in the pulse-mode insufflator group,in which,we observed widened gaps and vacuoles among the myocytes at 180 minutes after the insufflation,while in the other group,such changes were not apparently detected. Conclusions CO2 pneumoperitoneum may injure the muscular tissues below the peritoneum in rats. Constant-pressure mode insufflator is helpful to decrease such an injury.

Objective To evaluate the effect of CO2 pneumoperitoneum with various pressures on the ultrastructure of the implanted lesions in rats model of endometriosis. Methods Endometriosis model was established in 30 SD rats by implanting self-endometria onto the peritoneum to form ectopic endometrial tissues. Then the animals were divided randomly into three groups (ten in each):control,10 mm Hg,and 20 mm Hg CO2 pneumoperitoneum groups (CO2 inflation lasted for 1 hour). One week after the procedure,the ectopic endometrial tissues were sampled for electron microscopy to determine the ultrastructure of cell organelle in the endometriosis glands and matrix cells. Results The electron microscopy showed that in the control group,the matrix cells lined up in order and the structure of the microvilli (MV) was normal; while in the 10 mm Hg and 20 mm Hg groups,the chromatin margination of glandular cells and mitochondria swelling were observed with reduced or vanished mitochondrial crista,the ultrastructural damages to the microvilli (reduced or vanished) were more apparent than the control,the cell junctions were widened or collapsed,and the matrix cells lined up disorderly. Compared to the 10 mm Hg group,ultrastructural damage to the glandular cells in the 20 mm Hg group was more severe. Conclusions CO2 pneumoperitoneum intervention can induce ultrastructural damage to endometriosis lesions. 20 mm Hg pressure of CO2 pneumoperitoneum can induce severer damage to the ectopic endometriosis tissues than 10 mm Hg.

Objective To evaluate the efficacy of laparoscopic treatment for interstitial tubal pregnancy. Methods A total of 12 cases of interstitial tubal pregnancy underwent laparoscopy in our hospital from 1993 April to June 2007. Among the patients,9 received tubal ligation using lasso (1-0 suture were used during the surgery and the fetal tissues were removed at the same time),1 underwent salpingotomy to remove the fetus,1 were treated by salpingectomy using stapling device,and 1 received injection of MTX (40 mg). Results The procedure was completed within 30-120 minutes. The intraoperative blood loss was ≤20 ml in all of the cases,except one who received salpingectomy (200 ml). In 10 patients,the level of blood hCG decreased to a normal range in 2 weeks,while the other two cases recovered in 4 and 8 weeks respectively postoperation. Conclusion Laparoscopic treatment is safe and effective for interstitial tubal pregnancy.