Objective:To analyze the reasons of misdiagnosed cases in the intraoperative frozen pathological diagnosis of breast disease and explore effective measures and practices for targeted and standardized quality control.Methods:A retrospective analysis was conducted on 2 421 cases of breast intraoperative frozen pathological examination performed at Renmin Hospital of Wuhan University from Apr. 2020 to Dec. 2021. The results of intraoperative frozen pathological examination were compared with postoperative pathological results to calculate the overall concordance rate and misdiagnosis rate. Pathological classification and causative analysis were performed for misdiagnosed cases. Standardized management was implemented to address the identified issues, and an analysis was conducted on 2248 cases from Feb. 2022 to Nov. 2023 to compare the overall concordance rate and misdiagnosis rate before and after management.Results:The comparison between intraoperative rapid frozen pathology diagnosis and postoperative paraffin pathology diagnosis showed that among the 2 421 breast specimens, there were 2 377 cases (98.18%) with concordant results and 44 cases of misdiagnosis, resulting in a misdiagnosis rate of 1.82%. The pathological types of 44 misdiagnosed cases were analyzed, among which 10 cases were lymph node metastatic carcinoma, 5 were lobar tumors, 3 were intraductal papillary tumors, 7 were carcinoma in situ and common hyperplasia each, and 4 were carcinoma in situ, sclerosing adenopathy, and other invasive carcinoma each. Through the analysis of causes, it was found that poor slide quality, the need for immunohistochemistry assistance, careless slide reading, missed critical lesions, and other factors contributed to misdiagnosis, with variations in the causes of misdiagnosis among different pathological types. After implementing standardized management, the overall concordance rate significantly improved (98.93%) and the misdiagnosis rate significantly decreased (1.07%) .Conclusions:Intraoperative frozen pathological diagnosis is of great significance in the treatment of breast diseases. The targeted standardized quality control can help early detection and solve problems, reduce the differences between different doctors and technicians, and improve the accuracy of intraoperative frozen pathological diagnosis.

Objective:To analyze the reasons of misdiagnosed cases in the intraoperative frozen pathological diagnosis of breast disease and explore effective measures and practices for targeted and standardized quality control.Methods:A retrospective analysis was conducted on 2 421 cases of breast intraoperative frozen pathological examination performed at Renmin Hospital of Wuhan University from Apr. 2020 to Dec. 2021. The results of intraoperative frozen pathological examination were compared with postoperative pathological results to calculate the overall concordance rate and misdiagnosis rate. Pathological classification and causative analysis were performed for misdiagnosed cases. Standardized management was implemented to address the identified issues, and an analysis was conducted on 2248 cases from Feb. 2022 to Nov. 2023 to compare the overall concordance rate and misdiagnosis rate before and after management.Results:The comparison between intraoperative rapid frozen pathology diagnosis and postoperative paraffin pathology diagnosis showed that among the 2 421 breast specimens, there were 2 377 cases (98.18%) with concordant results and 44 cases of misdiagnosis, resulting in a misdiagnosis rate of 1.82%. The pathological types of 44 misdiagnosed cases were analyzed, among which 10 cases were lymph node metastatic carcinoma, 5 were lobar tumors, 3 were intraductal papillary tumors, 7 were carcinoma in situ and common hyperplasia each, and 4 were carcinoma in situ, sclerosing adenopathy, and other invasive carcinoma each. Through the analysis of causes, it was found that poor slide quality, the need for immunohistochemistry assistance, careless slide reading, missed critical lesions, and other factors contributed to misdiagnosis, with variations in the causes of misdiagnosis among different pathological types. After implementing standardized management, the overall concordance rate significantly improved (98.93%) and the misdiagnosis rate significantly decreased (1.07%) .Conclusions:Intraoperative frozen pathological diagnosis is of great significance in the treatment of breast diseases. The targeted standardized quality control can help early detection and solve problems, reduce the differences between different doctors and technicians, and improve the accuracy of intraoperative frozen pathological diagnosis.

Objective To investigate the clinical efficacy of Jiedu Huayu Granules(mainly composed of the Chinese herbal medicines of Artemisiae Scopariae Herba,Rhei Radix et Rhizoma,Hedyotis Diffusae Herba,Paeoniae Radix Rubra,Curcumae Radix and Acori Tatarinowii Rhizoma)in the treatment of liver failure(LF)and to observe their effects on endoplasmic reticulum stress and inflammatory response.Methods From May 2022 to May 2024,a total of 102 patients with LF of toxin-heat-stasis stagnation syndrome were collected from Hepatology Department of Yulin Traditional Chinese Medicine Hospital,and were divided into the control group and the study group by using a table of random numbers,with 51 cases in each group.The two groups were given comprehensive western medicine treatment for inhibiting inflammatory response and protecting liver,regulating intestinal microecology,counteracting viral infection,decreasing transaminase level and treating jaundice.Additionally,the study group was treated with Jiedu Huayu Granules.The course of treatment for the two groups covered eight weeks.Before and after treatment,the two groups were observed in the changes of traditional Chinese medicine(TCM)syndrome scores,biochemical indicators,inflammatory factors and endoplasmic reticulum stress indicators.After treatment,the clinical efficacy and the incidence of adverse reactions of the two groups of patients were compared.Results(1)After eight weeks of treatment,the total effective rate of the study group was 86.27%(44/51)and that of the control group was 68.63%(35/51),and the intergroup comparison(tested by chi-square test)showed that the efficacy of the study group was significantly superior to that of the control group(P<0.05).(2)After treatment,the TCM syndrome scores in both groups were significantly decreased compared with those before treatment(P<0.01),and the decrease in the study group was significantly superior to that in the control group(P<0.05).(3)After treatment,the serum levels of biochemical indicators of total bilirubin(TBIL),total bile acid(TBA),alanine transaminase(ALT),and aspartate transaminase(AST)in the two groups were significantly decreased compared with those before treatment(P<0.05),and the decrease in the study group was significantly superior to that in the control group(P<0.05 or P<0.01).(4)After treatment,the serum levels of inflammatory factors of interleukin 6(IL-6),tumor necrosis factor α(TNF-α),transforming growth factor β(TGF-β),and C-reactive protein(CRP)in the two groups were significantly decreased compared with those before treatment(P<0.05),and the decrease in the study group was significantly superior to that in the control group(P<0.01).(5)After treatment,the levels of endoplasmic reticulum response indicators such as glucose-regulated protein 78(GRP78),CCAAT enhancer-binding protein homologous protein(CHOP),and cysteine aspartic acid specific protease 12(caspase-12)in the two groups were significantly decreased compared with those before treatment(P<0.05),and the decrease in the study group were significantly superior to those in the control group(P<0.01).(6)The incidence of adverse reactions in the study group was 15.69%(8/51)and that in the control group was 17.65%(9/51),and the comparison between the two groups showed that the difference was not statistically significant(P>0.05).Conclusion The combined use of Jiedu Huayu Granules with comprehensive western medicine treatment exerts certain efficacy in treating patients with LF of toxin-heat-stasis stagnation syndrome.The combined therapy is effective on relieving clinical symptoms and improving liver function,and is helpful for relieving endoplasmic reticulum stress and inflammatory response.

Objective:To explore the effects of human chorionic gonadotropin (hCG), progesterone as well as the change of estradiol 12 h after hCG trigger on the outcomes of in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET). Methods:A retrospective study was conducted at the Center for Reproduction and Genetics of the Affiliated Suzhou Hospital of Nanjing Medical University. A total of 2 506 patients received IVF/ICSI-ET treatment with gonadotropin-releasing hormone agonist (GnRH-a) protocol from March 2015 to June 2020 were selected. With Spearman rank correlation analysis and path analysis, we explore the relationship among the changes of these hormones and the baseline characteristic of patients, as well as the relationship among the changes of these hormones and the outcomes of IVF treatment.Results:The increase of hCG was accompanied by the rise of progesterone and the decline of estradiol change rate ( r=0.094, P<0.001; r=-0.093, P<0.001). Meanwhile the rise of progesterone was accompanied by the decline of estradiol change rate ( r=-0.089, P<0.001). The dosage of hCG trigger was directly positively correlated to hCG level after hCG trigger, path coefficients (PC) was 0.307 ( P<0.001). Body mass index (BMI) was directly negatively correlated to hCG level and progesterone level after hCG trigger (PC=-0.434, P<0.001; PC=-0.154, P<0.001), whereas positively correlated to estradiol change rate (PC=0.097, P<0.001). Meanwhile the duration and dosage of gonadotropin (Gn) used were positively correlated to progesterone level after hCG trigger (PC=0.102, P<0.001; PC=0.080, P=0.030). hCG level and progesterone level after hCG trigger had positive correlation to oocyte retrieved rate (PC=0.098, P<0.001; PC=0.080, P<0.001). While estradiol change rate was not correlated to oocyte retrieved rate ( P>0.05). Progesterone level on hCG trigger day negatively related to normal fertilization rate (PC=-0.050, P=0.039). hCG level, progesterone level and estradiol change rate after hCG trigger had no correlation with high-quality embryo rate, clinical pregnancy rate and live birth rate (all P>0.05). Conclusion:Oocyte retrieved rate was positively affected by hCG level and progesterone level 12 h after hCG trigger. While normal fertilization rate, high-quality embryo rate, clinical pregnancy rate and live birth rate were not affected by the change of hormones level 12 h after hCG trigger. Therefore we should pay attention to hCG level and progesterone level 12 h after hCG trigger.

Objective:To study the feasibility of magnetic resonance imaging(MRI)technique with amide proton transfer(APT)in predicting the prognosis of cerebral stroke.Methods:A total of 71 patients with acute cerebral stroke who admitted to the Nanjing First Hospital,Nanjing Medical University from September 2022 to May 2023 were selected.All of them underwent the test of National Institute of Health Stroke Scale(NIHSS),and received the MRI examination with chemical exchange saturation transfer(CEST).According to the modified Rankin scale(mRS)values of 1-month follow-up,they were divided into favorable recovery group(mRS<2,44 cases)and poor group(mRS≥2,27 cases).The asymmetric magnetization transfer ratio(MTRasym)image(APT)was obtained by analyzing data with special software.And then,the difference(△APTw)of APT values between ischemic zone and contralateral normal tissue was further calculated.The △APTw values of two groups were compared and analyzed,and the Pearson correlation analysis was adopted to analyze the correlation among △APTw,NIHSS and mRS.The receiver operating characteristics(ROC)curve was drawn,and the area under curve(AUC)of ROC curve was calculated.Results:There were significant positive correlations among △APTw,NIHSS and mRS scores(R2=0.659,0.522,P<0.001),and the differences of △APTW,NIHSS and mRS scores between the favorable recovery group and poor group were significant(t=5.73,6.36,13.92,P<0.05),respectively.The AUC value was 0.886,and the sensitivity and specificity of prediction were respectively 77.8%and 95.5%.The positive and negative predictive values were respectively 91.3%and 87.5%.Conclusion:APT imaging technique has feasibility in predicting the prognosis of acute cerebral ischemic stroke.

OBJECTIVE To investigate the regulatory effect of autophagy on the resistance of human liver cancer cell Huh7 to lenvatinib. METHODS Using human liver cancer cell Huh7 as subject， the lenvatinib-resist cell model （Huh7-LR） was generated by the low-dose gradient method combined with long-term administration. The sensitivity of parental cell Huh7 and drug-resistant cell Huh7-LR to lenvatinib was detected by using CCK-8 assay and flow cytometry. Western blot assay and GFP-mCherry-LC3 plasmid transfection were performed to detect the expression levels of autophagic protein Beclin-1， autophagic adapter protein sequestosome 1 （p62）， microtubule-associated protein 1 light chain 3 （LC3） and autophagic level. Furthermore， an autophagy activation model was constructed by cell starvation， the protein expression of p62 and autophagy level were detected by using Western blot assay and GFP-mCherry-LC3 plasmid transfection， and the effect of autophagy activation on the sensitivity of Huh7-LR cells to lenvatinib was detected by flow cytometry. RESULTS Compared with parental cells， the drug resistance index of Huh7-LR cells was 6.2； protein expression of p62 was increased significantly， while apoptotic rate， protein expression of Beclin-1 and LC3Ⅱ/ LC3Ⅰ ratio were all reduced significantly （P＜0.05 or P＜0.01）； the level of autophagy was decreased to some extent. Autophagy activation could significantly increase the protein expression of p62 in Huh7-LR cells （P＜0.05） and autophagy level， and significantly increase its apoptotic rate （P＜0.05）. CONCLUSIONS Autophagy is involved in lenvatinib resistance， and activating autophagy can reverse the resistance of liver cancer cells to lenvatinib to some extent.

Objective:To analyse the clinical significance of selective single embryo transfer by time-lapse mo-nitoring(TLM)or conventional morphology assessment(CMA)in vitro fertilization/intracytoplasmic sperm in-jection and embryo transfer(IVF/ICSI-ET),and to initially explore the predictive value of Raman spectral analy-sis of embryo culture medium for clinical pregnancy rate.Methods:The study is a prospective randomized con-trolled clinical trial.We assigned 139 patients treated with IVF/ICSI-ET in Reproductive and Genetics Center of Suzhou Municipal Hospital from April 2019 to July 2020,which were randomly assigned to either the CMA or the TLM group.We performed selective single-embryo transfer(fresh cycle and FET)after selecting the optimal em-bryos with TLM or CMA respectively.If the patient's first embryo transfer was unsuccessful,a second one would be performed to compare the differences in the cumulative live birth rate of embryo transfer and other pregnancy outcomes between the two groups.Meanwhile,we collected 15 μl of embryo culture medium at day 3 after IVF/ISCI fertilization for Raman spectroscopy analysis.Results:There were no differences in cumulative live birth,cu-mulative clinical pregnancy,cumulative premature birth,cumulative early spontaneous abortion,cumulative ectopic pregnancy and LGA or SGA between TLM and CMA groups(P＞0.05).The Neonatal sex ratio in the TLM group was lower than that in the CMA group,but the difference was not significant(P＞0.05).Raman spectros-copy analysis of embryo culture medium predicted the clinical pregnancy rate with 67.21％accuracy.Conclu-sions:In young women with a good ovarian reserve,the advantage of using TLM to evaluate embryos is not obvi-ous,so we should remain vigilant that embryo selection based on morphokinetic parameters may affect the sex ratio.Raman spectroscopic analysis of embryo culture medium is not yet able to effectively predict the planting ability of embryos.

Objective:To explore the effects of human chorionic gonadotropin (hCG), progesterone as well as the change of estradiol 12 h after hCG trigger on the outcomes of in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET). Methods:A retrospective study was conducted at the Center for Reproduction and Genetics of the Affiliated Suzhou Hospital of Nanjing Medical University. A total of 2 506 patients received IVF/ICSI-ET treatment with gonadotropin-releasing hormone agonist (GnRH-a) protocol from March 2015 to June 2020 were selected. With Spearman rank correlation analysis and path analysis, we explore the relationship among the changes of these hormones and the baseline characteristic of patients, as well as the relationship among the changes of these hormones and the outcomes of IVF treatment.Results:The increase of hCG was accompanied by the rise of progesterone and the decline of estradiol change rate ( r=0.094, P<0.001; r=-0.093, P<0.001). Meanwhile the rise of progesterone was accompanied by the decline of estradiol change rate ( r=-0.089, P<0.001). The dosage of hCG trigger was directly positively correlated to hCG level after hCG trigger, path coefficients (PC) was 0.307 ( P<0.001). Body mass index (BMI) was directly negatively correlated to hCG level and progesterone level after hCG trigger (PC=-0.434, P<0.001; PC=-0.154, P<0.001), whereas positively correlated to estradiol change rate (PC=0.097, P<0.001). Meanwhile the duration and dosage of gonadotropin (Gn) used were positively correlated to progesterone level after hCG trigger (PC=0.102, P<0.001; PC=0.080, P=0.030). hCG level and progesterone level after hCG trigger had positive correlation to oocyte retrieved rate (PC=0.098, P<0.001; PC=0.080, P<0.001). While estradiol change rate was not correlated to oocyte retrieved rate ( P>0.05). Progesterone level on hCG trigger day negatively related to normal fertilization rate (PC=-0.050, P=0.039). hCG level, progesterone level and estradiol change rate after hCG trigger had no correlation with high-quality embryo rate, clinical pregnancy rate and live birth rate (all P>0.05). Conclusion:Oocyte retrieved rate was positively affected by hCG level and progesterone level 12 h after hCG trigger. While normal fertilization rate, high-quality embryo rate, clinical pregnancy rate and live birth rate were not affected by the change of hormones level 12 h after hCG trigger. Therefore we should pay attention to hCG level and progesterone level 12 h after hCG trigger.

Objective:To explore the effects of cyclosporine A (CsA) on the clinical outcomes of patients with unexplained repeated implantation failure (URIF) undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Methods:A retrospective cohort study was conducted, and the data of URIF patients at the Center for Human Reproduction and Genetics of the Affiliated Suzhou Hospital of Nanjing Medical University from April 2016 to March 2020 was analyzed. Totally 94 cycles with CsA application were enrolled, and 188 cycles of control group were selected according to age, body mass index (BMI), basal follicle-stimulating hormone (FSH), the number of high-quality embryos transferred and embryo stage matched with CsA group. The general conditions, characteristics of previous failed transfer cycles and the de novo transfer cycles, clinical outcomes, obstetric and neonatal complications were compared between the two groups, and regression analysis of CsA and clinical outcomes. Results:1) There were no statistical differences between the two groups in baseline characteristics such as age, BMI, basal FSH, duration of infertility, indications for IVF/ICSI, number of previous failed cycles, number of high-quality embryos and blastocyst transferred rate (all P>0.05). There were also no significant differences in the number of embryos transferred, the number of high-quality embryos, the rate of transferred blastocyst and the endometrial thickness on transfer day of the de novo transfer cycles (all P>0.05). 2) The embryo implantation rate, the clinical pregnancy rate and the live birth rate in CsA group [53.39% (63/118), 58.51% (55/94), 45.74% (43/94)] were distinctly higher than those of control group [38.43% (93/242), 45.74% (86/188), 33.51% (63/188)]. The differences were statistically significant ( P=0.007, P=0.043, P=0.046). While there were no differences in miscarriage rate and ectopic pregnancy rate between the two groups (all P>0.05). 3) The differences of the rates of preterm birth, multiple pregnancy, obstetric and neonatal complications were all not statistically significant between the two groups (all P>0.05). No birth defects were observed in both groups. 4) The multivariate logistic regression analysis showed that CsA was an independent promoter of clinical pregnancy (after adjusted OR=1.694, 95% CI:1.019-2.816, P=0.042) and live birth (after adjusted OR=1.700, 95% CI:1.012-2.853, P=0.045) in URIF patients after adjusting for age, BMI, basal FSH, the number of embryos transferred and the endometrial thickness on transfer day. Conclusion:CsA showed remarkably enhancement on embryo implantation rate, clinical pregnancy rate and live birth rate of de novo embryo transfer in patients with URIF, without increasing the risk of obstetric and pediatric complications. CsA application may be used as an effective treatment for URIF patients.

Abstract: Objective To clarify the long-term evolution of hepatitis B virus (HBV) quasi-species in HBsAg asymptomatic carriers in Long'an county, Guangxi. Methods ELISA was used to detect serological markers of HBV. Viral loads were measured by real time PCR. HBV DNA was extracted from serum by kits. The whole HBV genome was amplified using nested PCR and amplicons were sequenced by next-generation sequencing (NGS). These sequences from NGS were analyzed by the software like Mega. Results Serum samples were collected from 9 HBsAg asymptomatic carriers in Longan County，Guangxi at 4 different time points in 2004, 2007, 2013, 2019 or 2020. A total of 23 serum samples and 309 full-length gene quasi-species sequences were obtained, with an average amount of (0.18±0.07) G sequencing data for each sample. Genotype of 55.54%(5/9) the studied subjects underwent genotype conversion during the long-term evolution process of HBV quasi-species, and the genotyping results of the phylogenetic tree in the PreS/S region are in perfect agreement with the results of the whole genome analysis; recombinant B/C, I/C were found; the Sn ranged from 0 to 0.37 and the genetic diversity ranged from 0 to 0.11, respectively. A total of 21 special single nucleotide/amino acid mutations (7 in the S region, 2 in the X region, 3 in the PreC region and 9 in the BCP region) and 6 deletion mutations were detected, multiple mutations were found and no drug resistant mutations were found; 77.8%(7/9) of the HBV strains carried by the subjects in 2004 had double mutations at nt1 762(A→T) and 1 764(G→A) and a stop mutation at nt1 896(G→A); HBV mutations can be restored from the mutant type to the wild type and (or) vice versa without antiviral drug pressure, and The evolution rate of HBV genome was 2.03×10-5~3.50×10-3.Conclusion HBV genotype, recombinants, genetic complexity and diversity of HBV quasi-species can change over time during in natural infection. The transformation between HBV mutation type and wild type reduces the value of predicting clinical outcomes by genetic types and related mutations to some extent. The HBV genome evolution rate of asymptomatic carriers of HBsAg in Long'an County is very high.