Objective To detect the expression pattern of microRNA in A549 cells treated by lipopolysaccharide, study the expression of miRNA-1247-3P in A549 cells under LPS treatment and explore the possible mechanism of miRNA-1247-3P in A549 cells under LPS treatment. Methods A549 cells were divided into experimental and control groups. Immunocytochemical method and RT-PCR were used to detect the changes of SP-A and SP-C. The expression of miRNAs were detected using miRNAs array in different groups. The key miR-1247-3P was collected to detect the changes of miR-1247-3P in all groups using quantitative real-time polymerase chain reaction. Results Compared with control group, the expressions of SP-A and SP-C were significantly decreased in the experimental groups (P < 0.05). MiRNA array showed that 31 miRNAs were up-regulated and 3 miRNAs were down-regulated. Compared with control group, the expression of miR-1247-3P was significantly increased in the experimental groups (P < 0.05). Conclusion The increased expression of miR-1247-3P may play an important role in the pathogenesis of ARDS.

Objective To study the influence of progesterone on the cell proliferation of hormone?dependent breast cancer and observe the co?effect of vitamin D and progesterone of different levels on the proliferation of the cell line T?47D in hormone?dependent breast cancer and the interaction be?tween vitamin D and progesterone. Methods The cultured T?47D cells were divided into the high and the low progesterone mono?treated groups to observe the effects of different levels of progesterone on the cell proliferation of hormone?dependent breast cancer cell line(T?47D). The high and the low vitamin D mono?treated groups were set to observe the effects of different levels of vitamin D on the cell proliferation. Groups treated by differ?ent levels of progesterone combined with vitamin D were set to observe the interaction between them. Normal breast cancer cells were set as the con?trol group. The two?factor two?level parallel factorial experiment was conducted to observe the co?effect of different levels of progesterone and vitamin D on the proliferation of T?47D. The growth and apoptosis of cells was observed through detection of absorbance in each group by MTT. Results The cell concentration in high and low progesterone treated groups was increased than that in the control group,and was increased in the low proges?terone treated group compared with the high progesterone treated group(P<0.05). The cell concentration in dual?low level treated group was de?creased than that in the other three groups which were two?factor treated and the control group(P<0.05). Conclusion Progesterone stimulates the cell proliferation of hormone?dependent breast cancer. The concentration?based interaction between vitamin D and progesterone indicates that the bi?directional effect of progesterone on breast cancer cells may be related to the concentration of progesterone and other factors,but the specific interac?tion and the mechanism is unclear.

Objective:To detect the value of utilizing bpMRI in prostate biopsy in the detection of prostate cancer with PSA≤20ng/ml.Methods:The clinical data of 394 patients who underwent prostate biopsy in the First Affiliated Hospital of Nanjing Medical University from November 2017 to October 2019 were retrospectively analyzed. Of all the patients, 177 underwent modified systematic biopsy, named TRUS group, 217 patients accepted pre-biopsy bpMRI examination, undergoing modified systematic biopsy if Prostate Imaging Reporting and Data System (PI-RADS) score < 3 or MRI-TRUS cognitive fusion targeted prostate + systematic biopsy if PI-RADS score ≥ 3, named MRI group. The median age of TRUS group was 66 (61, 74) years old, prostate specific antigen (PSA) was 9.52 (7.26, 12.30) ng / ml, and prostate volume (PV) was 36.84 (28.95, 57.72)ml. The median age of MRI group was 66 (59, 72) years old, PSA was 8.84 (6.65, 12.16) ng/ml, and PV was 39.45 (29.25, 58.69)ml. There was no difference in above parameters between the two groups. The χ 2 test was used to compare the detection rate of prostate cancer and clinically significant prostate cancer (CsPCa) between the two groups. Results:There was no significant difference in the detection rates of prostate cancer between TRUS group and MRI group [51.41% (91/177) vs. 48.39% (105/ 217), P = 0.550], but the detection rates of CsPCa were significantly different [26.55% (47/177) vs. 36.41% (79/217), P = 0.037]. In patients with PSA ≤ 10 ng / ml, there was no significant difference in the detection rates of prostate cancer between the two groups [43.62% (41/94) vs. 43.08% (56/130), P = 0.936], but there was a significant difference in the detection rates of CsPCa [17.02% (16/94) vs. 28.46% (37/130), P = 0.047]. There was no significant difference in the detection rates of prostate cancer [60.24% (50/83) and 56.17% (48/87), P= 0.504] and the detection rates of CsPCa [37.35% (31/83) vs. 48.28% (42/87), P = 0.150] between the two groups. The total detection rates of the last two needles in TRUS group and MRI group were 23.16% (41/177) and 36.63% (86/217), respectively, with significant difference ( P=0.001); the detection rates of CsPCa in the last two needles were 11.86% (26/177) and 29.03% (63/ 217), respectively, with significant difference ( P < 0.001). In MRI group, the detection rates of prostate cancer in patients with PI-RADS score <3, 3, 4, 5 were 21.21% (7/33), 25.84% (23/89), 73.24% (52/71), 95.83% (23/24), respectively; the detection rates of CsPCa were 12.12% (4/33), 17.98% (16/89), 54.93% (39/71), 83.33% (23/24), respectively. Conclusions:In patients with PSA ≤ 20 ng / ml, prostate biopsy based on bpMRI may improve the detection of CsPCa, especially in patients with PSA ≤ 10 ng/ml.

BACKGROUND:Foreign scholars have researched hypoxia reperfusion in zebrafish embryos, but there is no research on c-fos gene expression and the mechanism during zebrafish cerebral hypoxia reperfusion.
OBJECTIVE:To observe the zebrafish embryonic brain cel apoptosis and expression of c-fos gene in brain tissues after hypoxia reperfusion.
METHODS:Zebrafish embryos were selected at 48 hours post fertilization. Neonatal hypoxia reperfusion injury was simulated by gradual y leading nitrogen (99.999%) into the device. After hypoxia treatment for 6, 12 and 24 hours, the embryos received reperfusion for 6 hours under normal oxygen concentration. The embryos in the control group received normoventilation (the dissolved oxygen concentration was about 7.0 mg/L). Acridine orange staining was performed to observe the effect of different hypoxia durations on the apoptosis of neurons in zebrafish, and then the c-fos gene expression was quantitative analyzed with real-time quantitative nucleic acid amplification detection system. And the expression level of c-fos gene was compared before and after hypoxia reperfusion.
RESULTS AND CONCLUSION:A smal amount of apoptotic brain cel s could be detected in the control group, and the c-fos gene expression level was decreased;in the experimental group, the number of apoptotic cel s was increased after hypoxia for 6, 12 and 24 hours, and the gene expression after hypoxia for 6 hours was increased distinctly. The results indicate that hypoxia can increase the c-fos gene expression in brain cel s of zebrafish embryos, which may be one of the mechanisms of brain cel apoptosis increasing after hypoxia.

Objective To investigate the effects of medicine androgen deprivation therapy(ADT)on quality of life in patients with prostate cancer.Methods A total of 42 consecutive advanced prostate cancer patients without any other anti-androgen medications after ADT[A subcutaneous depot injection of LHRH-agonist(Zoladex) was instituted every 28 days]were enrolled.Levels of serum testosterone and prostatic specific antigen(PSA)were obtained just prior to ADT and after ADT.The general and disease-specific health-related quality of life were assessed.Results The average testosterone level was less than 50 mg/L after medical ADT in 3 weeks.And PSA level declined dramatically in one month.Although there were no significant differences on physical discomfort and limitations to daily activities.Urinary obstruction symptoms after ADT were disappeared and the size of prostate were reduced after one month.The appetite and vigor were worsened.Overall health status and sexual function were significantly reduced.Conclusion The ADT could make serum testosterone and PSA decline dramatically in short time,and worsen some general health-related quality of life.

Objective To evaluate the protective effect of adiponectin in early atherosclerosis and the diagnostic value of adiponectin in metabolic syndrome in obese children. Methods Total 176 obese children and 88 normal weight children aged 9-12 years were included in the present study. All participants underwent hematologic and biochemical tests including serum adiponectin, high sensitivity C-reactive protein (hsCRP),fasting blood glucose, insulin, and plasma lipids. Homeostasis model assessment of insulin resistance (HOMA-IR)was calculated. Noninvasive ultrasound measurement including intima-media thickness of the common carotid artery(IMT), brachial flow-mediated dilatation (FMD), carotid artery compliance (CAC), and the maximum fatthickness ahead of peritoneum (Pmax) were obtained to investigate arterial mechanical properties and endothelial function. Results (1) The level of adiponectin was negatively correlated with obese index, blood pressure,fasting insulin, hsCRP, HOMA-IR, and IMT(P<0.05 or P<0. 01 ); but not with triglyceride, fasting blood glucose, CAC, high-density lipoprotein-cholesterol (HDL-C), and FMD. (2) The risk of metabolic syndrome increased 3.43 times in children with adiponectin level <7. 060 mg/L compared with ＞7. 060 mg/L. (3)Receiver operating characteristic( ROC ) curve was used to choose the optimal cutpoint of adiponectin to identify obese children with the metabolic syndrome. The area under the curve (AUC) for adiponectin to discriminate the sensitivity of metabolic syndrome was 0. 769 (95% CI0. 714-0.816, P< 0. 0 1 ). (4) The obese children were divided into three groups according to the cut-off value for adiponectin (high, middle, low groups). There were significant differences in the prevalences of severe obesity, visceral fat accumulation, hypertension, insulinemia,low HDL-C, metabolic syndrome among three groups (P<0.05). Conclusions High levels of serum adiponectin could prevent early stage of atherosclerosis. The lower the adiponectin level, the higher the incidence of metabolic syndrome.

Objective To explore the effects of the Chinese medicine, modified Erchen decoction, on the serum lipid spectrum of ApoE-/-mice, and to explore its possible anti-atherosclerotic mechanism.Methods Forty-four male 7-8-week old ApoE-/-mice were used in this experiment.ApoE-/-mouse models of atherosclerosis were generated by high-cholesterol diet for 4 weeks.And then, they were given simvastatin or modified Erchen decoction by gavage.The body weight of mice was recorded every week, The mice were sacrificed after treated with the drugs for 8 weeks continuously, and the plasma lipid was determined by enzymatic method.The aortic valves and arches were stained with oil red O to depict atherosclerotic plaques and liver structural changes of the mice were examined by pathology.Results Modified Erchen decoction lowered plasma lipid ( including TCHOL and LDL-C ) significantly ( P<0.01 ) .The body weight was increased in the mice of all groups, but it was more pronounced in the mice of model group than in the blank and modified Erchen decoction groups.The serum CHOL and LDL-C levels were significantly lowered in the modified Erchen decoction group (P<0.01).The area of atherosclerotic plaques in the aortic wall was significantly reduced in the mice of modified Erchen decoction group as shown by oil red O staining.The pathological changes of hepatocytes were less severe and the structure of hepatic lobules was better preserved in the mice of modified Erchen decoction group.Conclusions The Chinese medicin modified Erchen decoction can effectively reduce serum lipids, regulate lipid metabolism, and ameliorate the process of atherosclerosis in ApoE-/-mice.

Objective To discuss the mechanism of miR-24-3P in the process of acute respiratory distress syndrome (ARDS).Methods miR-24-3P mimics,miR-24-3P inhibitor and their negative controls were transfected in RAW264.7 cells respectively:The relative expression of miR-24-3P in each cell was detected.The relative expression of TNF-α,IL-6,SP1 and NF-κB were detected in each cell,and the regulation of miR-24-3P on the target gene SP1 was detected by dual luciferase reporter gene.Results The expression level of TNF-α mRNA and IL-6 mRNA in each group was statistically significant (P＜0.05).The relative expression level of SP1 mRNA and NF-κB mRNA in each group was not statistically significant (P＞0.05).The expression of SP1 in each group was statistically significant (P＜0.05).The gene analysis of dual luciferase reporter showed that the fluorescence activity was significantly inhibited after cotransfection of miR-24-3P and SP1 in HEK293 cells(P＜0.05).Conclusion miR-24-3P can play an important role in the process of ARDS by influencing the translation of SP1 gene and affecting the release of inflammatory mediators.

Objective:To investigate the relationship between the biochemical parameters, the pulmonary pathologic injury and the immune mechanism of severe sepsis in infant porcine, and the intervention effect of Shenfu injection.Methods:Panamanian infant porcine (2-3 months old) were divided into sham operation group (Sham group; intravenous injection of normal saline), lipopolysaccharide (LPS) induced severe sepsis model group (LPS group; intravenous injection of LPS 1 mg/kg, and continuing at 0.5 mg·kg -1·h -1 for 12 hours), and Shenfu injection intervention group (SF group; intravenous injection of Shenfu injection 10 mL/kg at the same time of modeling, twice a day) according to the random number table method, with 5 in each group. Forty-eight hours after the challenge, the changes of C-reactive protein (CRP), procalcitonin (PCT), oxygenation index (PaO 2/FiO 2), base excess (BE), blood lactate (Lac) and other biochemical indexes were detected with blood sampling; the number of neutrophils [myeloperoxidase positive (MPO +)] and their activated subsets CD11b + CD64 +, M1 macrophages (CD80 + CD64 +), CD4 + and CD8 + T cells were analyzed in peripheral blood by flow cytometry. The levels of plasma cytokines were detected by enzyme linked immunosorbent assay (ELISA). The pathological damage of lung tissue was observed by hematoxylin-eosin (HE) staining. The mRNA expression of lung injury related molecules and their receptors, chemokines, cytokines, vascular endothelial related molecules and tight junction protein were detected by reverse transcription-polymerase chain reaction (RT-PCR). Results:Compared with Sham group, the levels of CRP, PCT and Lac in LPS group significantly increased, and PaO 2/FiO 2 and BE significantly decreased. In the peripheral blood, CD80 + CD64 + macrophages, CD11b + CD64 + and MPO + neutrophils, CD4 +, CD8 + T cells and tumor necrosis factor-α (TNF-α) significantly decreased, but interleukin-10 (IL-10) and vascular endothelial cell growth factor (VEGF) significantly increased; the mRNA expressions of high mobility group protein B1 (HMGB1), receptor for advanced glycation end products (RAGE), angiopoietin 2/angiopoietin 1 (Ang2/Ang1) significantly increased, Toll like receptor 9 (TLR9), chemokines (CXCL9 and CXCL10), TNF-α, IL-27, Tek tyrosine kinase 2 (TIE2), vascular endothelial cadherin (VE-CAD) and Occludin significantly decreased in lung tissue. HE staining showed inflammatory cell infiltration and exudation in the alveolar cavity, alveoli consolidation, thickening of the alveolar interstitial layer and emphysema. Compared with LPS group, Lac in SF group significantly decreased (mmol/L: 4.2±1.0 vs. 6.3±1.1, P < 0.05), while BE and PaO 2/FiO 2 significantly increased [BE (mmol/L): -6.4±2.6 vs. -11.6±2.5, PaO 2/FiO 2 (mmHg, 1 mmHg = 0.133 kPa): 180±36 vs. 105±35, both P < 0.05]; the percentage of CD80 + CD64 + macrophages, CD11b + CD64 + and MPO + neutrophils significantly increased [CD80 + CD64 +: (7.13±2.01)% vs. (3.80±0.46)%, CD11b + CD64 +: (8.33±2.55)% vs. (2.15±0.47)%, MPO +: (21.22±2.33)% vs. (8.31±0.46)%, all P < 0.05]; the mRNA expressions of HMGB1 and RAGE decreased to some extent [HMGB1 mRNA (2 -ΔΔCT): 1.81±0.45 vs. 2.23±0.85, RAGE mRNA (2 -ΔΔCT): 6.69±3.48 vs. 11.60±6.91, both P < 0.05], the mRNA expressions of CXCL9 and TIE2 increased to a certain extent [CXCL9 mRNA (2 -ΔΔCT): 1.06±0.63 vs. 0.50±0.12, TIE2 mRNA (2 -ΔΔCT): 1.42±0.68 vs. 0.27±0.16, both P < 0.05]; the pathological damage of lung tissue were significantly alleviated. Conclusions:The increased abnormality of BE, Lac, and PaO 2/FiO 2, the immune exhausting and paralysis may be important factors leading to the fatal lung injury in infant porcine with severe sepsis. The possible mechanism is that the excessive activation of HMGB1 and its receptor RAGE, the suppression of TLR9 and corresponding chemokine and inflammatory factors lead to increased endothelial damage and decreased tight connection, which in turn induces capillary leakage. The intervention of immune disorders by Shenfu injection in the severe pneumonia accompanied by sepsis may be related to elevated M1 macrophages and activated neutrophils in the peripheral blood, inhibition of HMGB1 and its receptor RAGE mRNA expression, and elevated CXCL9 and TIE2 expression.

Objective To investigate the effect of cervical manual stratified anastomosis and anastomosis with tube stapler on the recent complications in thoracic laparoscopy combined with radical resection of esophageal cancer.Methods From February 2019 to April 2022,a total of 196 patients who underwent endoscopic surgery for esophageal cancer who met the study criteria were divided into the manual group(87 cases)and the tubulostomy group(109 cases)according to the different ways of gastro-esophageal cervical anastomosis.The incidence of cervical surgery time,total operation time,postoperative anastomotic fistula,anastomotic stenosis and other complications of the two groups were evaluated,and the differences in treatment effects between the two groups were compared.Results The preoperative basic conditions of patients in the manual group and the tube kiss group were comparable,and the cervical anastomosis time in the tube kiss group[(23±3.57)min]was shorter than that in the manual group[(31±4.5)min](P＜0.05),but there was no statistical significance in the overall operation time between the two groups(P＞0.05).The comparison of postoperative anastomotic fistula and anastomotic stenosis between the two groups showed that the manual group was significantly lower than the tube kiss group,and the difference was statistically significant(P＜0.05).Conclusion In thoracic laparoscopic combined with esophageal cancer surgery,cervical manual stratified anastomosis can reduce the incidence of postoperative anastomotic complications.