Objective:To investigate the mechanism of immunomodulatory effects of umbilical cord mesenchymal stem cells(UC-MSCs)modified by miR-125b-5p on systemic lupus erythematosus(SLE).Methods:The expression level of miR-125b-5p was detected by real-time fluorescence quantitative PCR in UC-MSCs and peripheral blood mononuclear cells(PBMCs)from SLE patients and health checkers.Annexin V-FITC/PI apoptosis detection kit was used to detect the effect of miR-125b-5p on apoptosis of UC-MSCs.MRL/lpr mice in each group were injected with UC-MSCs via tail vein,and T-lymphocyte subsets in the spleen of the MRL/lpr mice were detected by flow cytometry after 5 weeks.The expression levels of interleukin(IL)-4 and IL-17A in serum of MRL/lpr mice were detected by ELISA.Hematoxylin-eosin staining was used to observe the pathological manifestations of the lungs and kidneys of the MRL/lpr mice.Results:miR-125b-5p was significantly down-regulated in PBMCs of SLE patients compared with healthy controls(P＜0.01).Compared with the UC-MSCs group,the expression of miR-125b-5p in UC-MSCs modified by miR-125b-5p group was increased(P＜0.01).The survival rate of UC-MSCs was significantly increased by miR-125b-5p(P＜0.01).Compared with the untreated group of MRL/lpr mice,the expression level of IL-4 in serum was increased(P＜0.05);the expression level of IL-17A was decreased(P＜0.05);the proportion of Th17 cells in the spleen of MRL/lpr mice was decreased(P＜0.05);the inflammatory cells infiltration and micro-thrombosis of lungs and kidneys of MRL/lpr mice were significantly reduced in the UC-MSCs modified by miR-125b-5p treatment group.Conclusion:UC-MSCs modified by miR-125b-5p have immunomodulatory effects on systemic lupus erythematosus.

Feces and intestinal contents of pigs suspected with porcine epidemic diarrhea virus were collected from a farm in Minqin County,Gansu Province,China.After the suspected positive sam-ples were detected by RT-PCR,Vero cells were used to isolate and culture them in vitro.The suc-cessfully isolated virus was identified in the laboratory,and its whole genome sequence was ana-lyzed for genetic evolution.The pathogenicity was evaluated by animal regression test.The results showed that typical syncytial lesions could be observed when the PEDV-positive treatment solu-tion was inoculated with Vero cells in the 4th generation,and the virus titer in the 6th generation reached 10-4 75TCID50/mL.PEDV-like virions with a diameter of about 100 nm and a round shape with obvious capsular membranes and spikes were observed by electron microscopy.Whole genome sequencing analysis showed that the total length of this strain was 28 085 bp,which was far from the G1 subtype represented by the classical strain CV777(96.6％),and had a high homology with the G2b strains BC-2011-1,IA1,USA/Colorado/2013 and WELL(98.6％).This indicated that the strain belonged to the G2b epidemic strain.The animal regression test showed that the 5-day-old piglets developed vomiting,acute watery diarrhea,emaciation and mental depression within 12 h after the attack,and the symptoms worsened and died within 24 h.After autopsy,the infected piglets could be observed with stomach swelling,high intestinal heave,thin and transparent intesti-nal wall,and undigested milk clots inside.In summary,a PEDV G2b epidemic strain was success-fully isolated and identified in this study,and its whole genome sequence and pathogenicity were analyzed,providing research materials for future studies on PEDV gene function,pathogenic mech-anism and vaccine development.

Objective To standardize the selection of clinical research outcome indicators,which can objectively evaluate the clinical efficacy or effect of traditional Chinese medicine in the treatment of myasthenia gravis.This study aims to standardize the construction of the core outcome set of clinical research of traditional Chinese medicine in the treatment of myasthenia gravis.Methods We followed the core outcome set development specification(COS-STAD)to carry out research,established a research working group,which set up a Delphi-method advisory group.Two graduate students of working group conducted a document research and meetings of patients to establishe an outcome set item pool of myasthenia gravis in clinical trials of Chinese medicine under the instruction of other members.With the questionnaire based on the content of item pool,we then carried out Delphi-method expert consultations and a consensus meeting.Results The core outcome set of clinical research on myasthenia gravis treated with traditional Chinese medicine included five outcome domains:endpoint outcome,myasthenia gravis symptom evaluation,medication evaluation,quality of life evaluation and safety outcome;Nine outcome measures:recurrence rate,incidence of hormone complications,incidence of crisis,QMGS scale(MGFA quantitative myasthenia gravis score),daily activity scale of MG patients(ADL),analysis of immunosuppressant dosage,analysis of glucocorticoid dosage,analysis of cholinesterase inhibitor dosage,and incidence of adverse events.Conclusion The five outcome domains and nine outcome measures included in the core outcome set can be used as outcome options for the efficacy evaluation of myasthenia gravis clinical research.

Objective: This study aimed to investigate the impact of molecular classification and PTEN, KRAS and PIK3CA gene mutation on the outcome of fertility-preserving treatment in the patients with endometrioid endometrial cancer (EEC) and endometrial atypical hyperplasia (EAH). Methods: This is a single-center retrospective study. A total of 135 patients with EEC and EAH receiving fertility-preserving treatment and molecular classification were reviewed. The distribution of the four types of molecular classification was described. The impact of non-specific molecular profile (NSMP), mismatch repair-deficiency (MMRd), and PTEN, KRAS and PIK3CA gene mutation on the outcome of fertility-preserving treatment was analyzed. Results: Of the patients analyzed, 86.7% (117/136) were classified as having NSMP; 14 (10.4%), MMRd; 1 (0.7%), POLEmut EAH; and 3 (2.2%), p53abn EEC. The patients having NSMP and MMRd achieved similar 16-, 32-, and 48-week complete response rates. The patients harboring tier I and tier II PTEN mutations (PTENmut-Clin) achieved lower cumulative 32-week CR rates than those with PTEN-others (without PTENmut-Clin) (22/47, 46.8% vs. 50/74, 67.6%; p=0.023; odds ratio=0.422; 95% confidence interval [CI]=0.199–0.896). Insulin-resistance (hazard ratio [HR]=0.435; 95% CI=0.269–0.702; p=0.001) and PTENmut-Clin (HR=0.535; 95% CI=0.324–0.885; p=0.015) were independent negative predictors for lower 32-week CR rates. Conclusion PTENmut-Clin is an independent risk factor for unfavorable fertility-preserving treatment outcomes in the patients with EEC and EAH. The patients with MMRd receiving fertility-preserving treatment achieved outcomes similar to those of the patients with NSMP. The molecular profiles might guide fertility-preserving treatment in the prognosis and clinical decisions.

Objective:To explore the role of 17β-estradiol(17β-E2) derived from the hippocampus and gonad in the cognitive memory during the reconsolidation period of fear memory in rats with post-traumatic stress disorder(PTSD).Methods:The 8-week-old clean grade female SD rats were used for this study. Single prolonged stress combined with context fear conditioning was used to prepare the rat PTSD model.(1) Gonad estradiol experiment: 50 female rats were randomly divided into blank control group, model group, sham ovariectomy group, ovariectomy group, and ovariectomy+ estradiol group by random number table method, with 10 rats in each group. The rats in model group, sham ovariectomy group, ovariectomy group, and ovariectomy+ estradiol group were established PTSD model. Rats in ovariectomy group and ovariectomy+ estradiol group underwent ovariectomy, while rats in sham ovariectomy group only were removed adipose tissue of the same mass around the ovaries. And the rats in ovariectomy+ estradiol group were injected with 17β-E2(1 mg/kg, once a day for 14 days) 7 days after ovariectomy.(2) Hippocampal estradiol experiment: 40 female rats were randomly divided into blank control group, model group, hippocampal solvent control group and hippocampal estradiol inhibitor group by random number table method, with 10 rats in each group.The rats in model group, hippocampal solvent control group and hippocampal estradiol inhibitor group were established PTSD model.Rats in hippocampal estradiol inhibitor group were given letrozole once(bilateral, 0.5 μL per side) during the fear memory consolidation period) and rats in the hippocampus solvent control group were injected once with dimethyl sulfoxide in the hippocampus(bilateral, 0.5 μL per side) . The open field test and elevated cross maze test were used to evaluate the anxiety level and autonomous exploration ability, the freeze test was used to evaluate the fear memory, the new object recognition test was used to evaluate the non-spatial memory, and ELISA was used to detect serum 17β-E2 level. SPSS 25.0 software was used for data analysis, and one-way ANOVA was used for inter group comparisons, LSD test was used for further pairwise comparisons.Results:(1)In the gonad estradiol experiment, there were statistical differences in various indicators among the five groups of rats in the open field test, elevated cross maze test, freeze test, and new object recognition test( F=20.200, 12.702, 7.514, 10.094, 7.899, 13.211, all P<0.05). The number of upright times, central area activity time, distance and frequency of entering the open arm, and cognitive index of the model group rats were all lower than those of the blank control group(all P<0.05), and the freezing time was higher than that of the blank control group( P<0.05). The number of upright positions((11.20±1.55) times), central area activity time((11.33±1.80) s), distance((1.49±0.26) m) and times((10.00±1.50) times) entering the open arm, the freezing time((92.20±6.07) s) and cognitive index((60.40±3.71)%) in ovariectomy+ estradiol group were all higher than those of ovariectomy group((4.90±0.65) times, (4.31±1.07) s, (0.49±0.06) m, (3.10±0.62) times, (60.30±5.28) s, (32.60±8.08)%)(all P<0.05). (2) In the hippocampal estradiol experiment, there were statistical differences in various indicators among the four groups in the open field test, elevated cross maze test, freeze test, and new object recognition test( F=40.831, 5.553, 9.087, 5.848, 7.657, 9.191, all P<0.05). The numbers of upright positions, distance and frequency of entering open arms of the model group rats were lower than those of the blank control group(all P<0.05). The number of upright positions((3.00 ± 0.39) times), distance of entering open arm((1.17±0.37) m), freezing time((46.70±3.57)s), and cognition index((29.60±2.70)%) in the hippocampal estradiol inhibitor group were all lower than those in the hippocampal solvent control group((10.10±1.40) times, (4.02±0.79) m, (93.70±9.73) s, (54.20±5.08)%)(all P<0.05). Conclusion:17β-E2 derived from the hippocampus and gonad both had ameliorative effect on cognitive memory and anxiety-like behavior in the reconsolidation period of fear memory in PTSD model rats.

Hepatocellular carcinoma (HCC) is a common malignant tumor with poor prognosis and high mortality. In this study, we demonstrated a novel vaccine targeting HCC and tumor neovascular endothelial cells by fusing recombinant MHCC97H cells expressing porcine α-1,3-galactose epitopes (αGal) and endorphin extracellular domains (END) with dendritic cells (DCs) from healthy volunteers. END⁺/Gal⁺-MHCC97H/DC fusion cells induced cytotoxic T lymphocytes (CTLs) and secretion of interferon-gamma (IFN-γ). CTLs targeted cells expressing αGal and END and tumor angiogenesis. The fused cell vaccine can effectively inhibit tumor growth and prolong the survival time of human hepatoma mice, indicating the high clinical potential of this new cell based vaccine.

Objective:To preliminarily evaluate the efficacy and safety of stereotactic body radiotherapy (SBRT) for pulmonary oligometastatic tumors from head and neck carcinoma.Methods:Clinical data of 24 patients with pulmonary oligometastasis from head and neck carcinoma undergoing SBRT in Zhejiang Cancer Hospital from January 2014 to May 2019 were retrospectively analyzed. Survival analysis was performed by Kaplan- Meier method. Results:Among the 24 patients, 12 cases were diagnosed with nasopharyngeal origin and 12 cases of non-nasopharyngeal origin. A total of 34 pulmonary metastatic lesions were treated with SBRT. The median follow-up time was 19.5 months. Thirteen cases developed new lesions after SBRT, and 9 of them occurred within 1 year after SBRT treatment. The actual 1-year local control rate was 95%. The median progression-free survival was 15.2 months, and the 1-and 2-year progression-free survival were 59% and 46%, respectively. The 2-and 3-years overall survival rate at were 71% and 51% fter lung metastasis, respectively. Univariate analysis showed that the patients with primary lesions located in nasopharynx and disease-free interval of more than 1 year had survival advantage. No more than grade 3 radiation-induced injury was observed in the whole cohort after SBRT, and the incidence of mild radiation-induced injury was 13%.Conclusions:SBRT is safe and effective in the treatment of pulmonary oligometastatsis from head and neck carcinoma, and it may be more effective for patients with primary tumors located in nasopharynx.

Objective:To study the correlation between liver imaging reporting and data system (LI-RADS) category with tumor differentiation, Ki67 index, microvascular infiltration, and predictive prognosis in hepatocellular carcinoma (HCC).Methods:We retrospectively analyzed the clinical and radiological data of 178 patients with HCC who were confirmed by histopathological studies after liver resection between January 2015 and September 2020 at Lishui Central Hospital and Lishui People’s Hospital. There were 156 males and 22 females, with age of (57±10) years old. These patients were assessed for LI-RADS categories according to the 2018 version of LI-RADS, and they were divided into 4 groups according to the assessment results: 12 patients with LI-RADS-3 (the LI-RADS-3 group); 26 patients with LI-RADS-4 (the LI-RADS-4 group); 102 patients with LI-RADS-5 (the LI-RADS-5 group); and 38 patients with LI-RADS-M (the LI-RADS-M group). The patients' general information, tumor markers, pathology and other clinical data were recorded. Correlation analysis between the LI-RADS category with pathology was performed by the Kendall's tau-b test. Survival analysis between groups was performed by the Kaplan-Meier analysis. The Cox regression risk model was used to analyze the relationship between these variables with the risk of death.Results:The Kendall's tau-b test showed that LI-RADS category was positively correlated with the degree of tumor differentiation ( t=0.204, P=0.002), but not with microvascular infiltration and Ki 67 index ( P>0.05). All patients were followed up for 4.2 to 84.2 months (median follow-up 36.3 months). By the end of follow-up, 31 patients had died and 147 patients were alive. The cumulative 1-year and 3-year survival rates of the LI-RADS-5 group were 97% and 90% respectively, which were significantly higher than those in the LI-RADS-M group (81% and 63%), and the LI-RADS-4 group (96% and 81%), ( P<0.05). The cumulative 1-year and 3-year survival rates of patients in the LI-RADS-3 group were 100% and 67% respectively, and there was no statistically significant difference with the LI-RADS-5 group ( P>0.05). The Cox multivariate regression analysis showed that tumor glycoantigen 199 (>50 μl/ml) to be an independent influencing factor in survival of HCC patients ( HR=0.43, 95% CI: 0.24-0.76, P=0.004). Conclusion:The LI-RADS category of HCC was positively correlated with the degree of tumor differentiation, and patients with HCC meeting the LI-RADS-5 criteria had relatively better prognosis.

Objective: This study aimed to investigate the impact of polycystic ovary syndrome (PCOS) on fertility-sparing treatment in young patients with atypical endometrial hyperplasia (AEH) or endometrioid endometrial cancer (EEC). Methods: A total of 285 patients with EEC (n=76, FIGO stage IA, without myometrium invasion) or AEH (n=209) who received progestin-based fertility-sparing treatment were evaluated retrospectively. Among the 285 patients, 103 (36.1%), including 70 AEH cases and 33 EEC cases, were diagnosed with PCOS. General characteristics, cumulative 16- and 32-week complete response (CR) rate, pregnancy outcome and recurrence were compared between patients with or without PCOS. Results: The cumulative 16-week CR rate was lower in the PCOS group than in the non-PCOS group (18.4% vs. 33.8%, p=0.006). Patients with PCOS took longer treatment duration to achieve CR (7.0 months vs. 5.4 months, p=0.006) and shorter time to relapse after CR (9.6 months vs. 17.6 months, p=0.040) compared with non-PCOS group. After adjusting for patient age, body mass index, PCOS, homeostasis model assessment-insulin resistance index, and serum testosterone levels, we found that body mass index ≥25 kg/m2 (HR=0.583; 95% CI=0.365–0.932; p=0.024) and PCOS (HR=0.545; 95% CI=0.324–0.917; p=0.022) were significantly correlated with lower 16-week CR rate. Conclusion PCOS was associated with lower 16-week CR rate, longer treatment duration and shorter recurrence interval in patients with AEH or EEC receiving fertility-preserving treatment.

BACKGROUND: The cardiomyocyte apoptosis is considered as one of major contributions to cardiac remodeling after myocardial infarction (MI). Numerous studies find that circular RNAs (circRNAs) play pivotal roles in a variety of biological functions. However, the role of circ_0068655 in MI and human induced pluripotent stem-derived cardiomyocytes (HCMs) remains unknown. METHODS: The expression of circ_0068655, miR-498, and PRKC apoptosis WT1 regulator (PAWR) in human MI heart tissues and hypoxia subjected HCMs was evaluated with qRT-PCR and Western blot. The effects of circ_0068655 on hypoxia-induced apoptotic death and cell migration in HCMs were evaluated with qRT-PCR, cell viability, cell death ELISA (POD), and Caspase-3 activity assay, and Trans-well assay, respectively. Furthermore, luciferase assay, qRT-PCR, biotin-labeled miRNA pulldown assay, and Western blot were employed in the functional studies. RESULTS: We found that the expression of circ_0068655 and PAWR was enhanced in MI patients and hypoxia subjected HCMs; by contrast, the expression of miR-498 decreased. Inhibited expression of circ_0068655 in HMCs counteracted hypoxia-induced apoptotic death and impaired cell migration, in sharp contrast to circ_0068655 knockdown. We identified that circ_0068655 sponged an endogenous miR-498 to sequester and inhibit its activity, leading to the increased PAWR expression. CONCLUSIONS Our findings reveal that the expression of circ_0068655 can promote cardiomyocyte apoptosis through the modulation of miR-498-PAWR axis in vitro, which highlights the diagnostic and therapeutic value of circ_0068655 in patients with MI.