BACKGROUND:In vitro fertilization-embryo transfer is commonly used to solve infertility,but its success rate is not high,the more common reasons are poor endometrial receptivity,poor egg quality,etc.The follicular fluid melatonin can inhibit the aging of the ovary,to a certain extent,can promote the development of embryos,improve the probability of conception,but whether there is a correlation between the two is not known. OBJECTIVE:To explore the correlation between follicular fluid melatonin level and pregnancy rate of single-cycle in vitro fertilization-embryo transfer women. METHODS:A total of 112 female patients who received in vitro fertilization-embryo transfer treatment in the First Affiliated Hospital of Anhui Medical University from December 2020 to April 2021 were selected as the study subjects.They were divided into quartile array(Q1-Q5)according to the follicular fluid melatonin level from low to high.Among them,the melatonin level of group Q1 was<6.99 ng/L(n=18),that of group Q2 was 7.00-9.99 ng/L(n=26),that of group Q3 was 10.00-11.99 ng/L(n=27),and that of group Q4 was 12.00-13.99 ng/L(n=18);and melatonin levels in group Q5 were 14.00-19.99 ng/L(n=23).Clinical data characteristics of the five groups were compared.Multi-factor Logistic regression was used to analyze the correlation between follicular fluid melatonin level and pregnancy rate of women with single-cycle in vitro fertilization-embryo transfer and embryo transfer.A restricted cubic spline Logistic regression model was established to analyze the dose-response relationship,and the model was evaluated by clinical decision curve. RESULTS AND CONCLUSION:(1)Compared with the study population with the lowest melatonin quintile(Q1),with the increase of melatonin level(Q2-Q5),the levels of egg harvest and pregnancy success were gradually increased,and the body mass index was gradually decreased,and the differences were significant(P<0.05).(2)Multivariate Logistic regression analysis showed that after adjusting for confounding factors such as global mass index,number of eggs retrieved,luteinizing hormone,estradiol,progesterone and other confounding factors,follicular fluid melatonin level was still independently correlated with pregnancy rate of single-cycle in vitro fertilization-embryo transfer women(OR=1.538,95%CI:1.032-1.837,P<0.05),and there was significant difference in trend test of follicular fluid melatonin level from low to high quintile array(Ptrend<0.05).(3)The sensitivity test analysis showed that E value was 2.117.Subgroup analysis showed that the study population with higher levels of luteinizing hormone in follicular fluid had a more significant association between follicular fluid melatonin and pregnancy rate in single-cycle in vitro fertilization-embryo transfer women(P interaction=0.008).(4)The results of restricted cubic spline model analysis showed that there was a nonlinear dose-response relationship between follicular fluid melatonin level and pregnancy rate of single-cycle in vitro fertilization-embryo transfer women(P<0.05),and there was an overall positive correlation between follicular fluid melatonin level and pregnancy rate of single-cycle in vitro fertilization-embryo transfer women.(5)The results of clinical decision curve analysis showed that the follicular fluid melatonin level had important clinical value in predicting the pregnancy rate of single-cycle in vitro fertilization-embryo transfer women.(6)Follicular fluid melatonin level is closely related to the pregnancy rate of single-cycle in vitro fertilization-embryo transfer women,and with the decrease of follicular fluid melatonin level,the pregnancy rate of single-cycle in vitro fertilization-embryo transfer women also decreases.

BACKGROUND:The clinical manifestation of acute radiation skin injury is recurrent necrotic ulcers,and its pathogenesis is still not fully understood.The establishment of a suitable animal model will have important clinical implications for the study of its pathogenesis,prevention and treatment. OBJECTIVE:To establish a model of acute β-ray radiation skin injury and to investigate the mechanism of injury. METHODS:Sixty-nine Sprague-Dawley rats were randomly divided into 30,45,60 Gy 32P-β-ray groups(n=21 per group)and control group(n=6).A single local irradiation of the back of the rats was performed using 32P radionuclide.The control group was operated in the same way as the irradiated groups except that it was not irradiated.The body mass and skin appearance of the rats were measured at 7,15,30,45,and 60 days after irradiation.Three rats from each group were selected at each observation time point.The skin injury was observed by hematoxylin-eosin staining,Masson staining,transmission electron microscopy,and TUNEL assay.P53,Bcl-2 and Bax protein levels in the skin were measured by immunohistochemistry and western blot assay. RESULTS AND CONCLUSION:There was no accidental death after irradiation,and the body mass of rats showed a gradual increase.The rats showed different degrees of epidermal necrosis,inflammatory cell infiltration,reduction of hair follicles and appendages,and collagen fibrillation,which were evident at 60 and 45 Gy.The levels of serum inflammatory factors,interleukin-6 and tumor necrosis factor-α,were significantly increased in a dose-dependent manner.Under the electron microscope,there are varying degrees of mitochondrial reduction,vacuolization and nuclear pyknosis in the cells.The degree of cell apoptosis showed a certain dose-dependence.Immunohistochemistry and western blot results showed an increase in the expression of P53 and Bax proteins and a decrease in the expression of Bcl-2 protein in the skin after irradiation.There were significant differences between the 60 Gy group and the 45 Gy and 30 Gy groups(P＜0.05).To conclude,irradiation with 60 Gy and 45 Gy 32P radionuclide on the back of rats could successfully establish a practically pre-clinical animal model,and the mechanism is related to the up-regulation of P53 and Bax and the down-regulation of Bcl-2.This model can provide a reference for the establishment of animal models for the study of the mechanism of radiation skin injury and its prevention and treatment.

BACKGROUND:The research of dental stem cells in the fields of regenerative medicine and tissue engineering has been deepening,bringing hope for the repair of tooth-related tissues and the treatment of systemic diseases.However,there is a lack of systematic research and analysis on the biological characteristics of dental stem cells in different age groups. OBJECTIVE:To explore the biological characteristics of the human deciduous tooth and permanent tooth pulp stem cells cultured in umbilical cord blood platelet lysate to provide a reliable basis for human platelet lysates to replace fetal bovine serum. METHODS:The pulp tissues of deciduous teeth,juvenile permanent teeth and adult permanent teeth were taken out and cultured in DMEM/F-12 medium supplemented with 10%fetal bovine serum or different concentrations(5%,10%and 15%)of human platelet lysates.Cell proliferation in the four groups was detected by cytometry.The optimal concentration of human platelet lysates was selected for subsequent experiments.Under the optimal concentration of human platelet lysates,human deciduous tooth and juvenile and adult permanent tooth pulp stem cells were cultured in vitro.The cell growth status was observed under the microscope.The specific antigen on the cell surface was detected by flow cytometry.The cell proliferation ability was tested by the cell counting method and CCK-8 assay.The cell differentiation ability in vitro was observed by a three-line differentiation assay. RESULTS AND CONCLUSION:(1)The cell proliferation rate of the 10%human platelet lysate group was the highest.(2)In all three groups,fusiform fibrous cells grew and expanded from around the tissue block.There was no significant difference between deciduous teeth and juvenile permanent tooth cells,but the adult permanent tooth cells were larger than the deciduous and juvenile permanent tooth cells of the same generation.(3)The results of flow cytometry showed that deciduous teeth,juvenile permanent teeth and adult permanent teeth conformed to the phenotypic characteristics of mesenchymal stem cells.(4)The proliferative capacity of adult permanent dental pulp stem cells was significantly lower than those of deciduous teeth and juvenile permanent dental pulp stem cells(P<0.01).(5)mRNA expressions of osteoblast-related genes alkaline phosphatase and bone morphogenetic protein 2,lipoprotein lipase and peroxisome proliferator-activated receptor γ2,mRNA expressions of chondroblast related gene type II collagen α1 and cartilage oligomeric matrix protein in adult pulp stem cells of permanent teeth were significantly lower than those of deciduous teeth and juvenile permanent teeth pulp stem cells(P<0.01).(6)Compared with adult dental pulp stem cells,human deciduous teeth and juvenile permanent teeth dental pulp stem cells have the stronger proliferative capacity and multidirectional differentiation potential,and are more suitable for clinical research and disease treatment.

In order to establish a rapid,specific and sensitive detection method for Streptococcus equi subspecies zooepidemicus(SEZ)and to understand the infection status of SEZ in horses ente-ring China,specific primers were designed and synthesized based on the conserved gene comB of standard strain SEZ(ATCC 43079)in this work.Then,the pMD19-T-comB recombinant plasmid was constructed and used as a standard positive template.After that,the fluorescence-based quantitative PCR(qPCR)detection method based on SYBR Green Ⅰ dye was established.Totally,477 equine entry serum samples from 6 countries,including Netherlands,Belgium,Japan,Germa-ny,Argentina and New Zealand,during 2018 to 2023,were randomly selected and detected for SEZ by the qPCR method.Results showed that the established qPCR method had specific amplification for only SEZ,which illustrated a good specificity.Sensitivity test of the method showed that the limited detection amount was 4.58 X101 copies/μL.And the repeatability test showed that the coef-ficient of variation of intra-batch repeatability was less than 0.5％,while the inter-batch repeat-ability was less than 3.0％,which indicated good repeatability and high stability.Retrospective a-nalysis showed that totally 11 of 477 positive samples were detected,with a relatively low positive rate of 2.31％(11/477).Among them,all the 40 samples from Netherlands in 2018 were negative(0/40).In the samples of 2019,one positive was detected from Belgium(1/20),while all other 36 samples which form Japan and Germany were negative.In the samples of 2021,three samples(3/34)from Japan and one sample(1/20)from Argentina were positive,and all the other 40 samples from the Netherlands were negative.In the samples of 2022,76 samples from Netherlands were all negative.While in the 2023,5(5/126)of 126 samples from Netherlands and one(1/88)of 88 from New Zealand were found positive with SEZ.To summarize,The SYBR Green Ⅰ qPCR method for the diagnosis of SEZ was successfully established,and it could provide necessary technical support for the rapid quarantine of China's entry-exit and port departments,as well as the epidemiological investigation of the disease.

Objective To investigate the relationships between the expression levels of tumor necrosis factor receptor associated factor 4 (TRAF4) and ribosomal S6 protein kinase 4 (RSK4) protein in gastric cancer tissues and the recurrence after laparoscopic radical gastrectomy. Methods In total, 176 patients were divided into the recurrence and non-recurrence group, and the expression levels of TRAF4 and RSK4 protein in cancer and adjacent tissues and in gastric cancer tissues in the recurrence and non-recurrence group were compared. The influencing factor of recurrence and the efficacy of TRAF4 and RSK4 protein expression in predicting recurrence were analyzed. Results The positive expression rate of TRAF4 protein in gastric cancer tissues was higher than that in adjacent tissues (P < 0.05) and that in the recurrence group was higher than that in the non-recurrence group (P < 0.05). The positive expression rate of RSK4 protein in gastric cancer tissues was lower than that in adjacent tissues (P < 0.05) and that in the recurrence group was lower than that in non-recurrence group (P < 0.05). The largest tumor diameter 5 cm, poor differentiation, TNM Ⅲ stage, depth of invasion T3-T4, lymph node metastasis, absence of adjuvant chemotherapy after operation, positive expression of TRAF4 and RSK4 protein, and regular diet w influenced the post-operative recurrence (all P < 0.05). The accuracy of TRAF4 and RSK4 protein in gastric cancer tissues in combined predicting the recurrence was 83.52%. Conclusion The expression of TRAF4 protein is high, and the RSK4 protein is low in gastric cancer tissue, which are related to recurrence.

Objective:To explore the clinical and pathological characteristics and prognostic factors of gastric neuroendocrine carcinoma(G-NEC)and gastric mixed adenoendocrine carcinoma(G-MANEC).Methods:Retrospective analysis was conducted on the clinical data of 67 patients with G-NEC and G-MANEC who underwent surgical treatment at Heping Hospital Affiliated to Changzhi Medical College from May 2015 to May 2023.The study included an analysis of the pathological characteristics distinguishing G-NEC from G-MANEC.Results:Com-pared to gastric adenocarcinoma,patients with G-NEC and G-MANEC in the stomach showed a higher incidence of gastric cancer in the male gastric cardia and were diagnosed at a later age.Tumors with larger diameters increase susceptibility to anemia,low albumin levels,and in-vasion of nerves and vasculature.Deeper tumor infiltration is associated with increased local lymph node metastases,later TNM staging,and a higher likelihood of distant metastasis post-surgery.The prognosis of G-NEC and G-MANEC is worse than that of gastric adenocarcinoma(P=0.001).However,there is no statistically significant difference in the pathological characteristics(P>0.05)and prognosis analysis(P=0.212)between G-NEC and G-MANEC.Univariate survival analysis identified age,preoperative albumin,preoperative CEA,number of lymph node metastases,TNM staging,and postoperative distant metastasis as risk factors affecting patient's overall survival(OS).In the multivariate ana-lysis,age,preoperative albumin,TNM staging,and postoperative distant metastasiswere identified as independent risk factors for OS.Con-clusions:There is a significant difference in clinical characteristics between G-NEC,G-MANEC,and gastric adenocarcinoma,often diagnosed at an advanced stage,which is prone to distant metastasis post-surgery.Poor prognosis is observed in patients aged over 60 years,with pre-operative albumin<40g/L,TNM stage Ⅱ/Ⅲ,and postoperative distant metastasis.

Stroke is one of the most common cerebrovascular diseases， including hemorrhagic stroke and ischemic stroke. From a modern medical perspective， stroke is caused by cerebrovascular damage or embolism leading to impaired blood circulation. From the traditional Chinese medicine （TCM） perspective， the pathogenesis of this disease is mainly due to the disorder of Qi and blood， which ascend to the brain， causing either blood extravasation or blockage of brain collaterals. Stasis is a pathological factor that runs throughout the entire course of stroke， and the method of promoting blood circulation and resolving stasis has been a core treatment for stroke for a long time. Hirudo， as a traditional insect drug， has shown good effects in promoting blood circulation and resolving stasis. Modern pharmacological research has confirmed that Hirudo contains anticoagulant components， which provide significant advantages in dissolving thrombi in ischemic stroke and facilitating hematoma absorption in hemorrhagic stroke. Hirudo and its related preparations have been proven to exert an anti-stroke effect through anticoagulation， anti-thrombosis， and protection of vascular endothelium. As a result， they have been widely used in the treatment of stroke. This article explored the theoretical basis and research status of using Hirudo for treating stroke based on its main active components and hemostatic properties and summarized the current research status of commonly used Hirudo-based formulations and preparations， aiming to provide references for the involvement of Hirudo in stroke treatment.

Objective : To analyze the pregnancy outcome of preimplantation genetic testing for aneuploidy (PGT-A) in cycles with different indications． Methods : The clinical information of 549 couples who underwent PGT-A were retrospectively analyzed．The cycles were divided into 6 groups according to the indication for PGT-A，namely : recurrent pregnancy loss group(n = 304) ，repeated implantation failure group(n = 57) ，advanced age group( ≥38 years old，n = 80) ，history of adverse pregnancy group(chorionic trisomy or adverse pregnancy，n = 24) ，male factor infertility group (n = 67) ，and abnormal sex chromosome number group (n = 17) ．The basic information，the number of retrieved oocytes，embryo biopsy result and pregnancy outcome were compared among different indication groups. Results: The average age and days of gonadotropin ( Gn) used among the six groups were statistically different (P＜0. 001) ．The average number of retrieved oocytes，the rate of good-quality embryos，mosaic embryos， abnormal embryos and normal embryos，the average ovarian sensitivity index ( OSI) among the six groups were statistically different (P = 0. 03，P ＜0. 001，P = 0. 03，P ＜0. 001，P ＜0. 001，P ＜0. 001 ) ．Advanced age group had the highest rate of abnormal embryos，the least average number of retrieved oocytes，the lowest OSI and the lowest rate of normal embryos．There were statistical differences in clinical pregnancy rate，ongoing pregnancy rate and cumulative pregnancy rate per oocyte retrieved cycle (P＜0. 001) among the six groups，but there were no statistical differences in clinical pregnancy rate，ongoing pregnancy rate per transfer cycle and cumulative pregnancy rate among the five groups except for the male factor infertility group． Conclusion PGT-A can detect euploid embryo to transfer thereby improving pregnancy efficiency．The advanced age women have normal embryo to transfer and can obtain a better pregnancy rate，which may shorten their time of“take- baby -home ”．At the same time， PGT-A can significantly improve the pregnancy outcome of those with male factor infertility.

Objective To analyze the epidemiological characteristics of automatic warning signals of infectious diseases received in Tianjin during the 13th Five-Year Plan period, and to provide reference for the function optimization of automatic warning system of infectious diseases during the 14th Five-year Plan period． Methods The data of infectious disease automatic early warning signals in Tianjin from 2016 to 2020 were downloaded from the China infectious diseases automated-alert and response system (CIDARS). The characteristics and response of early warning signals were analyzed by descriptive method. Results During the 13th Five-Year Plan period, 23,660 early warning signals were received in Tianjin, involving 39 kinds of infectious diseases, with 100% response rate and 1.10% positive rate of early warning signals. The median response time of warning signal was 0.53 hours (P₂₅-P₇₅: 0.19-1.17h). 95.74% of signals were responded within 2 hours and 98.47% of signals were responded within 24 hours. COVID-19, other infectious diarrhea diseases, dysentery, measles, and scarlet fever were the top five diseases with early warning signals. Conclusion During the 13th Five-Year, the automatic early warning system of infectious diseases in Tianjin runs well and has a high response rate, but the positive rate of early warning signal is low. During the 14th Five-Year，further research is needed to improve the early warning threshold according to the prevention and control needs and epidemic characteristics of different diseases, and to improve early warning positive rate.

Objective:To analyze the laboratory detection methods and clinical characteristics of patients with 2019-nCoV Omicron variant infection, to realize the rapid identification and diagnosis of 2019-nCoV Omicron variants.Methods:Totally 80 overseas patients in First Hospital of Changsha from December 16 in 2021 to January 5 in 2022 were selected, the nucleic acids and mutant genes were detected by fluorescent PCR and genome sequencing, and the clinical characteristics of patients with 2019-nCoV Omicron variant infection were analyzed.Results:The specificity was 100% (58/58) and positive predictive value was 100% (21/21) respectively, the sensitivity was 95.5% (21/22), negative predictive value was 98.3% (58/59) by detected with fluorescent PCR. It was found that there were 45-50 nucleotide displacement sites in the genome and 25-30 amino acid mutation sites in S gene fragment by genome sequencing. Clinical analysis showed that mild cases were 59.1% (13/22) in layouts, without severe and critical cases. Ages were positively associated with the clinical classification (ρ=0.698, P<0.001), foundation infections were positively associated with the clinical classification (ρ=0.636, P<0.001). Conclusions:Patients with 2019-nCoV Omicron variant infection had a high viral load and long negative conversion time of nucleic acid. Ages and foundation infections were positively associated with the clinical classification. AST/ALT was higher in the early stage of the disease. Fluorescent PCR method can be used in rapid screening patients with 2019-nCoV Omicron variant infection.