Abstract:Objective：This study aims to investigate the effects and mechanism of urocalum on the formation of kidney calcium oxalate stones. Methods：A rat model with renal calcium oxalate stone was constructed. Urocalum was intervened in this model for four weeks and renal samples were collected for HE staining，while urine samples were collected for the examination of biochemical components. In addition，autophagy and apoptotic key proteins were detected in renal tissues. A model of calcium oxalate stone cells was constructed by calcium oxalate crystallization in tubular epithelial cells. The cell model was treated with urocalum to explore the protective effect and molecular mechanism of urocalum on renal tubular epithelial cells. Results：The number of calcium oxalate stones was significantly increased compared to the control group. Whereas，urocalum could significantly inhibit the formation of stones，which may be associated with reducing oxidative stress levels，autophagy and apoptosis levels. In the cell model，α-sulphpoic acid can significantly inhibit the renal cell oxidative stress process induced by calcium oxalate crystallization，and both cell autophagy and apoptosis levels were activated after oxidative stress. Urocalum can also inhibit the oxidative stress process，while the phosphorylation of MPAK signaling pathway was significantly reduced and levels of autophagy and apoptosis were remarkably lowered. Conclusion：Urocalum can significantly inhibit the formation of renal calcium oxalate stones，and attenuate the oxidative stress process induced by calcium oxalate crystallization，and significantly reduce autophagy and apoptosis through the MAPK signaling pathway.

Abstract:Objective：This study aims to investigate the effects of progesterone on the expression of NLRP3 and the differentiation of Th17/Treg in murine splenocytes infected by Neisseria gonorrhoeae（Ng）. Methods：Murine splenocytes were isolated in vitro，and randomly divided into five groups：control group，Ng group，Ng+1×10-9 mol/L progesterone group，Ng+1×10-8 mol/L progesterone group，Ng+1×10-7 mol/L progesterone group. The splenocytes were collected after 3 days of culture，the proportions of Th17/Treg cells were detected by intracellular staining using flow cytometry. The mRNA expressions of NLRP3，Th17/Treg-specific transcription factors RORγt and Foxp3 in murine splenocytes were detected by real-time PCR. The protein expression levels of NLRP3，Caspase-1p20，RORγt and Foxp3 in murine splenocytes were determined by Western blot. The concentrations of interleukin（IL）-1β and Th17/Treg related cytokines IL-17，transforming growth facfor（TGF）-β，IL-10 in the supernatant were detected by ELISA. Results：Under the stimulation of Ng，the proportions of Th17 cells and Treg cells，the mRNA and protein levels of NLRP3，Th17/Treg-specific transcription factors RORγt and Foxp3 in the splenocytes，the protein levels of Caspase-1p20 and the concentrations of IL-1β，IL-17，TGF-β and IL-10 in the supernatant increased than those in the controls（P < 0.01）. After pretreated with progesterone，the proportion of Th17 cells，the mRNA and protein levels of NLRP3 and RORγt in the splenocytes，the protein levels of Caspase-1p20 and the concentrations of IL-1β and IL-17 in the supernatant were significantly lower than those in Ng group. The proportion of Treg cells，the mRNA and protein levels of Foxp3 in the splenocytes and the concentrations of TGF-β and IL-10 in the supernatant were significantly higher than those in Ng group（P < 0.01）. Conclusion：Progesterone can inhibit the expression of NLRP3 and Th17 differentiation，and promote the differentiation of Treg cells in murine splenocytes induced by Ng infection.

Abstract:Objective：This study aims to investigate the influence of silanes with different end groups grafting on the zirconia surface on the adhesion，proliferation and differentiation of osteoblast． Methods：A self-assembled monolayer was formed on the surface of zirconia by covalent bonding. After hydroxylation，zirconia surface was grafted with different terminal groups of silanes，and then mouse preosteoblasts are cultured on the surface. X-ray photoelectron spectroscopy（XPS），scanning electron microscope（SEM）and X-ray diffraction（XRD） analysis were used to detect the physical and chemical characteristics of the zirconia surface. The adhesion and morphology of MC3T3-E1 cells were observed by laser confocal microscope after 24 h culture. After 1，3 and 5 days，the cell proliferation level of each group was evaluated by the CCK-8 assay，and the differentiation of osteoblasts after 7 and 14 days of culture was detected by the alkaline phosphatase kit. Results：XPS results showed that the four different terminal silanes were successfully grafted to the zirconia surface. SEM and XRD results indicated that alkali heat and silane treatment had no obvious impact on the topography and crystal phase of zirconia surface. Zirconia grafted with hydrophilic terminal amino- and mercapto- silanes，presented excellent adhesion and high proliferation activity，and the expression of alkaline phosphatase is higher than that of the untreated group，while the hydrophobic methyl- and vinyl- silanes has no obvious effect on cell proliferation，and inhibit cell differentiation. Conclusion：Zirconia modified with hydrophilic end groups（such as mercapto and amino）of silanes can promote cell adhesion，proliferation and differentiation.

Abstract:Objective：This study aims to use bioinformatics methods to screen the differentially long non-coding RNA（LncRNA） in vaginal epithelial tissues of women with vaginal lubrication disorder（LD），and to construct patients with vaginal lubrication disorder based on the hypothesis of lncRNA-mRNA co-expression network to further explore the potential pathogenesis of patients with vaginal LD，in order to provide new ideas for the diagnosis and treatment of patients with vaginal LD. Methods：Using |Log2FC|≥2 and correct P ralue＜0.05，the expression profile of long-chain non-coding RNA and mRNA between LD group and normal control group were identified. Using Cytoscape software，a lncRNA-mRNA network of candidate lncRNAs was constructed. Using Gene Ontology（GO）and KEGG Pathway，the biological functions of mRNAs in co-expression network were analyzed. Results：A total of 499 up-regulated lncRNAs，337 down-regulated lncRNAs，and 1 582 up-regulated mRNAs，633 were selected in the LD group and the normal control group based on the next-generation sequencing technology according to the criteria of |Log2FC|≥2 and correct P ralue＜0.05. Subsequently，a lncRNA-mRNA co-expression network was constructed based on differentially expressed lncRNA and mRNA through 100 lncRNAs and 311 mRNAs. Finally，the functional enrichment analysis of the lncRNA-mRNA network showed that mRNA was mainly related to myocardial-related diseases and functions. Conclusion：The pathogenesis of LD may be related to blood circulation dysfunction，local muscle dysfunction，and cGMP pathway，etc. This requires further studies at the cellular level，animal level and even human level to confirm.

Abstract:Objective：This study aims to investigate the effects of metformin on the content and composition of long-chain fatty acids（LCFA） in pregnant women with gestational diabetes mellitus（GDM） and their newborns. Methods：A total of 127 pregnant women who delivered in Changzhou Maternal and Child Health Hospital Affiliated to Nanjing Medical University from August 2018 to August 2020 were selected，32 pregnant women with normal glucose tolerance（N Group）and 32 pregnant women with GDM who only needed diet control （G1 Group）were selected as control. According to the principle of randomization and voluntariness，the pregnant women with GDM who needed drug treatment were randomly divided into metformin treatment group（G2 group，33 cases） and insulin treatment group（G3 group，30 cases）. Blood glucose，blood lipids and insulin resistance index（HOMA-IR） were compared before delivery in each group. Body mass index（BMI）before pregnancy and delivery，and pregnancy outcome were compared. The composition and content of long LCFA in maternal blood and umbilical cord blood were analyzed. Results：The weight gain during pregnancy in G2 group was lower than that in N group，G1 group and G3 group. Fasting blood glucose（FBG）in G2 and G3 group was higher than that in N group. The 2-hour postprandial glucose（2hPG）in G1，G2 and G3 group was higher than that in N group. HOMA-IR in G2 group was higher than that in N group，G1 group and G3 group（P＜0.05）. There were no statistically significant differences in total cholesterol，triglyceride，free fatty acid in maternal blood and neonatal weight among the four groups（P > 0.05）. The main 11 kinds of LCFA in maternal blood and cord blood were analyzed by liquid chromatography，which were eicosenoic acid（EA），docosapentaenoic acid（DPA），linolenic acid（LA），α-linolenic acid（ALA），arochidonic acid（AA），docosahexaenoic acid（DHA），phosphatidic acid（PA），oleic acid（OA），eicosapentenoic acid（EPA），w-6-docosapentaenoic acid（DPA6），gamma linolenic acid（GLA）. EA in G2 and G3 groups was higher than that in G1 group，while DPA in G2 group was higher than that in N group，G1 and G3 groups（P < 0.05）. The contents of EA，DPA，LA，ALA，PA，OA，EPA，DPA6 and GLA in umbilical cord blood were lower than those in maternal blood（P < 0.05）. The levels of EA，LA，ALA，AA，DHA，PA，OA，EPA and GLA in maternal blood were significantly negatively correlated with FBG（P < 0.05）. Conclusion：Metformin and insulin have the same effect in the control of neonatal weight，blood glucose and blood lipid in the treatment of GDM，and metformin is better than insulin in the control of pregnancy weight. It has no significant effect on the contents of important fatty acids such as LA and ALA in maternal blood and umbilical cord blood，and has no significant effect on the contents of AA and DHA in umbilical cord blood. From the perspective of effect on essential fatty acids，the use of metformin in GDM is safe.

Abstract:Objective：This study aims to investigate the mutation of homologous recombination repair（HRR）associated germline genes and the correlation between clinical characteristics and mutation status in breast cancer. Methods：The mutation status of 21 HRR associated genes was detected by next-generation sequencing in 102 patients diagnosed with breast cancer in our center. The clinicopathological data of the patients was collected including the age，tumor size，lymph node status，pathological type and family tumor history. The association between the clinical pathological features and mutation status was analyzed. Results：There was no significant statistical difference between patients with mutation of HRR associated genes（BRCA or non-BRCA）and those without any mutation in clinical characteristics such as age，lymph node status，tumor size，pathological type and so on. Patients with BRCA mutation incline to have family history of tumor（95% CI：0.997-6.142，P=0.047），especially for family history of breast/ovarian cancer（95% CI：1.227-9.953，P=0.015）. Meanwhile，the mutation rate of ATM in human epidermal growth factor receptor-2（Her-2）positive patients was significantly higher than that in Her-2 negative patients（P=0.045）. Conclusion：The family history of tumor，especially of breast/ovarian cancer，is a vital risk factor for BRCA mutation；Her-2 positive breast cancer may be associated with ATM mutation.

Abstract:Objective：This study aims to clarify the effects of H19 genetic variant rs2839698，rs217727，rs3741216 and rs3741219 on renal cell carcinoma（RCC） susceptibility and prognosis. Methods：We conducted this two-stage case-control study with a total of 1 014 RCC cases and 1 063 controls since May 2004. Total 298 RCC cases in the first stage had complete follow-up data available. Genotyping was performed using TaqMan real-time polymerase chain reaction assays. Odds ratio（OR）and 95% confidence interval（CI） were calculated to evaluate the association between H19 polymorphism and RCC risk and clinical characteristics. Kaplan-Meier method was utilized to assess the survival of H19 polymorphisms. Hazard ratio（HR）and 95%CI were calculated by COX regression model to discover whether genotypes，TNM and grade were the independent prognostic factors. Results：Compared with the H19 rs2839698 CC genotype，the variant genotypes（CT/TT）were significantly associated with increased risk of RCC（P=0.012，OR=1.13，95% CI=1.02-1.55）. Besides，patients with variant genotypes（CT/TT）were more likely to develop large tumor（P=0.003，OR=1.35，95% CI=1.10-1.73）and advanced disease（P=0.010，OR=1.63，95% CI=1.08-2.21）；and had a significantly unfavorable 5-year survival than those with the rs2839698 CC genotype（CT/TT vs. CC：Log-rank P=0.027，HR=2.24，95%CI=1.10-4.59）. Conclusion：The results suggested that H19 rs2839698 variant may be a genetic predictor of susceptibility and mortality of RCC. The precise functional impact of the variant on H19 still needs further experimental validation.

Abstract:Objective：This study aims to screen and verify microRNA in neonatal acute respiratory distress syndrome （ARDS），preliminary study on miR-6833-3p level and its diagnostic effect in serum of ARDS neonate. Methods：Twenty-five infants with ARDS and 32 controls were investigated in this study. The miRNA microarray was utilized to explore the expression of miRNA in serum. Real-time PCR was used to verify the reproducibility of diagnostic chips and the miRNA target prediction on miRNAs that were over five folds differences between groups. Furthermore，correlation was conducted between levels of miR-6833-3p and APACHE Ⅱ scores. The diagnostic effect，sensitivity，and specificity were analyzed according to receiver operating characteristic（ROC） curve. Results：Twenty-four aberrant miRNAs were screened out as highly expressed，and among these miRNAs there were 8 miRNAs with over five folds differences of expression：miR-31-5p，miR-4754，miR-6833-3p and miR-192-3p were up-regulation；miR-362-3p，miR-11a，miR-7a-2-3p and miR-1382 were down-regulation. Through verification，it was found that the expression of miR-6833-3p in the plasma of the ARDS group was significantly up-regulated（P < 0.01） and positively correlated with the acute physiological score in APACHE Ⅱ score（r=0.731，P < 0.001）. The miR-6833-3p may be closely related to PI3-K/Akt and MAPK signaling pathways according to target gene prediction analysis. ROC curve showed that area under the curve was 0.848，with optimum threshold was 1.03，Youden’s index was 0.59 with sensitivity of 84.55% and specificity of 75.36%. Conclusion：The miR-6833-3p was significantly raised in neonatal ARDS，and may has certain clinical diagnostic efficacy as a new biomarker of neonatal ARDS.

Abstract:Objective：By observing and studying the specific related factors of patients with chronic obstructive pulmonary disease（COPD），a pre-hospital screening model of COPD was established to improve the implementation rate of pulmonary function examination in high-risk groups of COPD，so as to reduce the rate of missed diagnosis and mortality of COPD. Methods：Total 68 patients with stable COPD and 40 patients in the control group were enrolled. Their basic data were collected，and COPD assessment test（CAT），pulse rate and finger pulse oxygen measurement during exercise and pulmonary function test were performed at the same time. Logistic regression analysis was used to determine the independent variables related to COPD，and COPD screening model was constructed，which was corrected by Hosmer-Lemeshow test and discriminated by area under receiver operating characteristic curve（AUROC）. Results：There were significant differences in age，smoking index，body mass index（BMI），CAT score，lowest finger pulse oxygen（SPO2L）and highest pulse rate between the control group and stable COPD patients. Smoking index，CAT score and SPO2L constituted the screening model，and used to estimate the probability of COPD（PCOPD）.The best threshold of PCOPD was 57.1%. The best thresholds of influencing factors in the screening model were smoking index ≥220 cigarette-years，CAT score ≥ 7.5 and SPO2L ≤ 93.5%. The AUROC of the model was 0.938 with strong discriminant ability . The model correction was good with the Hosmer-Lemeshow fitting test P=0.789. Conclusion：Smoking index，CAT score and SPO2L constitute the pre-hospital screening model for COPD，which has good differentiation and correction. When PCOPD ≥ 57.1%，it is helpful for doctors to effectively screen high-risk patients with COPD，and it is recommended to carry out lung function test to confirm the diagnosis， so as to reduce the rate of missed diagnosis of COPD. When smoking index ≥ 220 cigarette-years，CAT score ≥ 7.5 and SPO2L ≤ 93.5%，it has good guiding significance for COPD screening.

Abstract:Objective：This study aims to compare the vascular function of the descending branch of lateral circumflex femoral artery（DBLCFA）and the internal mammary artery（IMA）of coronary artery bypass grafts，according to the vasoconstrictor and diastolic effects of the two drug pharmacological response to evaluate its spasticity characteristics and endothelial function. Methods：The 33 descending branches of the external circumflex femoral artery and 31 internal mammary arteries of 20 patients with coronary artery bypass grafting were balanced in the organ bath. Vasoconstriction was detected by receptor-mediated vasoconstrictors endothelin-1，angiotensin Ⅱ and thromboxane A2 mimic U46619. Acetylcholine was used to test endothelium-dependent relaxation，and nitroglycerin was used to test endothelium-independent relaxation. The maximum contraction or relaxation response of the two arteries，and the effective concentration that caused 50% of the maximum contraction or relaxation response were compared. Results：In the contraction caused by endothelin-1，angiotensin Ⅱ and U46619，the reactivity of DBLCFA was higher than that of IMA，respectively［（0.94±0.09）g vs.（0.57±0.08）g，P=0.012；（0.88±0.13）g vs.（0.24±0.05）g，P < 0.001；（4.95±0.57）g vs.（1.86±0.30）g，P=0.001］. When converted into the percentage of contractile force caused by 100 mmol/L KCl，there was no significant difference between the two groups. There was no significant difference in the effective concentration of 50% of the maximum contractile response caused by endothelin-1 and angiotensin Ⅱ. The effective concentration of 50% of the maximum contractile response caused by U46619 of DBLCFA was higher than that of IMA. There were no significant differences between receptor-mediated endothelium-dependent relaxation and non-endothelium-dependent relaxation between the two types of blood vessels. There were no significant differences in the effective concentration of 50% of the maximum diastolic response between the two arteries. Conclusion：DBLCFA has a higher pharmacological response to vasoconstrictors than the IMA，suggesting that it is more prone to vasospasm. The diastolic response of DBLCFA is similar to that of IMA，but DBLCFA is more prone to spasm. So its vascular function is inferior to that of the IMA. But when the use of IMA is limited，DBLCFA may be an option for the surgeon.

Abstract:Objective：This study aims to evaluate the value of measurement of interleukin（IL）-36 in peritoneal dialysis effluent in predicting the peritoneal solute transport rate and prognosis in continuous ambulatory peritoneal dialysis（CAPD）patients. Methods：From June 2018 to December 2019，a total of 111 CAPD patients in peritoneal dialysis center of the First Affiliated Hospital of Soochow University were analyzed，and overnight peritoneal effluents and peripheral blood were collected. The patients were grouped into H/HA or L/LA group according to the peritoneal equilibration test（PET）results，and the clinical data of the patients were collected. All the patients were followed up for 17 months，and adverse events were recorded. ELISA assays were used to assess IL-36α，IL-36β，and IL-36γ levels in peritoneal effluents and serum. The predictive role of effluent IL-36 was determined by Pearson correlation and logistic regression analysis. Results：The IL-36α AR and IL-36γ AR levels in overnight drained dialysate were significantly correlated with D/P Cr（r=0.51，P＜0.001；r=0.55，P＜0.001）and D/D0 glucose （r=-0.40，P＜0.001；r=-0.44，P＜0.001）. Logistical regression analysis showed that the effluent IL-36α AR or IL-36γ AR level in peritoneal effluents was an independent predictive factor for the peritoneal solute transport rate（OR=1.40，P=0.001；OR=1.08，P=0.016）. The area under the receiver operating characteristic（ROC）curve of effluent IL-36α AR was 0.75（95%CI：0.66-0.84，P < 0.001）；IL-36γ AR was 0.79（95%CI：0.70-0.88）. Moreover，effluent IL-36α AR was also an independent predictive factor for the adverse events of CAPD patients（OR=1.21，P=0.006），and the ROC curve of IL-36α AR was 0.76（95%CI：0.64-0.89）. Conclusion：Effluent IL-36 maybe a potential biomarker for predicting peritoneal solute transport rate and prognosis of CAPD patients.

Abstract:Objective：This study aims to investigate the pregnancy outcome of pregnant women with moderate and severe thrombocytopenia in perinatal pregnancy. Methods：the clinical data of 214 pregnant women with thrombocytopenia who delivered in the obstetrics department of the First Affiliated Hospital of Nanjing Medical University from January 2016 to December 2020 were analyzed retrospectively. They were devided into mild thrombocytopenia group（158 cases） and the pregnancy with moderate to severe thrombocytopenia group（56 cases）. Results：There was no significant difference in body mass index，pregnancy times，labor times and history between the pregnancy with mild thrombocytopenia group and the pregnancy with moderate to severe thrombocytopenia group（P > 0.05）. However，the age of pregnant patients with moderate to severe thrombocytopenia was older（P=0.03），and the gestational week of thrombocytopenia was nearly 7 weeks earlier than that of mild thrombocytopenia group（P < 0.001）. Excluding the influence of delivery mode on intrapartum bleeding，there were no significant differences in intrapartum bleeding between mild thrombocytopenia group and moderate to severe thrombocytopenia group（P > 0.05）. At the same time，the correlation between the severity of thrombocytopenia during pregnancy and postpartum hemorrhage and neonatal outcome was analyzed. There were no significant differences in the severity of thrombocytopenia during pregnancy，postpartum hemorrhage and neonatal outcome between two groups（P > 0.05）. In terms of the changes of coagulation factors before and after delivery，the changes of most coagulation factors in mild thrombocytopenia group were significantly higher than those in moderate to severe thrombocytopenia group（P < 0.05）. Conclusion：Although pregnant women with moderate to severe thrombocytopenia show thrombocytopenia，they should be comprehensively evaluated in combination with other coagulation related indicators，such as thromboelastogram and five coagulation items. Therefore，for pregnant women with thrombocytopenia，especially moderate to severe thrombocytopenia patients，reasonable and effective intervention should be carried out as soon as possible，and the mode of delivery should be selected individually to improve maternal and infant outcomes.

Abstract:Objective：This study aims to retrospectively analyze the effects of low and medium-dose glucocorticoids in treating of severe corona virus disease-2019 （COVID-19）. Methods：A retrospective analysis of the clinical data of 95 patients with severe COVID-19 admitted to the Huangshi Hospital of Traditional Chinese Medicine assisted by the Huangshi Medical Team of the Affiliated Sir Run Run Hospital of Nanjing Medical University from Feb，15th，2020 to Mar，20th，2020 was conducted. Control group（conventional treatment，33 cases）or hormone treatment group（conventional treatment combined glucocorticoid，62 cases）were divided according to whether using glucocorticoids or not. Laboratory test results，time of clinical symptoms relieved，the 28-day mortality rate，severe to critical convention rate，hospitalization days，inflammation index，improvement of oxygenation index，imaging changes and the occurrence of adverse reactions were compared between the two groups. Results：In general clinical data，the lymphocyte count and monocyte count from the hormone treatment group were significantly lower than that from the control group（P < 0.05），and there was no significant difference in the remaining baseline clinical data between the two groups. Compared with the control group，the clinical symptom relief time from hormone treatment group was significantly shorter［9.50（5.00，14.50）d vs. 16.00（5.00，21.00）d，P ＜ 0.001］，the conversion rate of severe to critical illness was significantly reduced［12.90% vs. 33.33%，P=0.029］，and the time of hospital stay was significantly shortened［20.00（16.00，22.00）d vs. 24.00（20.50，26.50）d，P=0.002］，the 28-day mortality rate was not increased［6.45% vs. 9.09%，P=0.691］. The oxygenation index recovery increased，the inflammatory factors of C-reactive protein and IL-6 decreased，and the imaging absorption speed increased significantly. There was no significant difference in adverse reactions between the two groups. Conclusion：Low and medium-dose glucocorticoids can significantly shorten the clinical symptom relief time of patients with severe COVID-19，improve oxygenation，promote lung lesion absorption，and have not obvious adverse reactions. It can be used in a short course during the treatment of severe COVID-19.

Abstract:The development of CRISPR-Cas9 technology has greatly promoted the progress of gene editing. Base editors and Prime editors developed based on this technology confer CRISPR-Cas9 system more powerful gene editing capabilities，and accelerate its application in clinical gene therapy. Although the CRISPR-Cas9 system has been optimized much，it still needs to be improved in terms of specificity，safety and delivery in vivo. This review briefly introduces and looks forward to the research progress regarding the above three concerns.

Abstract:Trained immunity（TI），independent of the T and B cells and distinct from the characteristic of traditional innate immune，is a memorial innate immune response. The organism can show increased responsiveness upon subsequent stimulation with the same or a different stimulus，well after the initial challenge. Trained immunity，as a new concept，involves the interaction of innate immune and the metabolic and epigenetic mechanisms. Based on this，this article summarized the effects of stimulation such as lipopolysaccharide（LPS），bacille calmette-guerin（BCG），and hormones on epigenetic and metabolic modifications of innate immune cells such as monocytes/macrophages，natural killer cells and non-classical immune cells，and explored the long-term vision of modulating training immunity in inflammation，nervous system diseases and COVID-19.

Abstract:Recent studies have found that long non-coding RNA（LncRNA）affect the occurrence and development of tumors by regulating genes related to cell proliferation and apoptosis. LncRNA ZEB2-AS1 regulates various aspects of tumor cell biological behaviors including proliferation，migration，invasion，apoptosis and glycolysis，both in vivo and in vitro. This review summarizes the research progress of the effect of ZEB2-AS1 on tumor biological behaviors and its working mechanisms，and evaluates whether it can be considered as a therapeutic target and biomarker for tumor diagnosis and prognosis.