Objective To investigate the effect of cyclosporine A (CsA) on autophagylysosomal pathway in tubular epithelial cells.Methods Human renal tubular epithelial cell line (HK-2 cell) was treated with different concentrations (3,5 and 10 μmol/L) of CsA for 24 h.Then the viability and apoptosis of cells were measured by MTT assay or AnnexinV-PI staining followed by flow cytometry analysis,respectively.Autophagy-related protein LC3-Ⅱ and p62 were detected by immunofluorescence assay.Autophagic flux was analyzed in HK-2 cells transfected with a tandem mRFP-GFP fluorescent-tagged LC3 (ffLC3) plasmid by laser confocal microscope.The lysosomal degradation was evaluated by DQ-ovalbumin staining followed by flow cytometry analysis.Results The viability of HK-2 cells was significantly decreased with CsA stimulation when compared with control group (P ＜ 0.01),but the number of apoptotic cells was markedly increased by CsA treatment (P ＜ 0.05).Compared with the control group,different doses of CsA dramatically increased the expressions of LC3-Ⅱ (P ＜ 0.01) and p62 (P ＜ 0.05) in HK-2 cells.Moreover,HK-2 cells treated with CsA displayed a significant increase in autophagosomes but a marked decrease in autolysosomes.In HK-2 cells,exposured to CsA caused a decrease in lysosomal degradation by DQ-ovalbumin staining when compared with control group (P ＜ 0.01).Conclusion Blockade of autophagy via disrupting lysosome degradation may represent a novel mechanism of CsA-induced tubular epithelial cells injury.

To reveal the anti-inflammation mechanism of koumine,we determined effects of different concentrations of koumine(100,200,400 mg/L) on the secretion of NO,IL-1β,IL-6 and TNF-a by nitrate reductase and ELISA as well as the mRNA of iNOS,IL-1β,IL-6 and TNF-α.Western blot was used to detect iNOS protein expression.The results showed that 100,200,400 mg/L of koumine can significantly inhibit the secretion of NO,IL-1β,IL-6 and TNF-α of RAW264.7 cell (P＜0.01).Koumine can dose-dependently down-regulate the mRNA of iNOS,IL-1β,IL-6 and TNF-α,meanwhile koumine can also significantly inhibit the protein expression of iNOS.The results indicated that koumine may play an anti-inflammation activity by mean of the reduction of NO and mediator of inflammation,down-regulating the mRNA expression of iNOS,IL-1β,IL-6 and TNF-a,decreasing protein expressions of iNOS.

AIM To investigate the tissue distribution of exendin-4 after administration in healthy rats. METHODS Exendin-4 was radioiodinated by the Iodo-GenTMmethod. Tissue distribution of [125I]exendin-4 was investigated after sc administration of [125I]exendin-4 at 3 μg·kg-1 in rats. Both total radioactivity and trichloroacetic acid (TCA) precipitated radioactivity were used to calculate the levels of [125I]exendin-4 in rats plasma and tissue samples after sc administration. RESULTS The tissue distribution of [125I]exendin-4 after sc injection showed substantial disposition in kidneys, lungs, bladder and pancreas. The rank order of normalized tissue distribution was kidneys＞lungs＞bladder＞pancreas＞intestine＞plasma＞adrenals>jejunum＞lymph＞liver＞spleen＞heart＞marrow＞thymus＞testicles＞brain＞muscle＞adipose. CONCLUSION [125I]Exendin-4 underwent a rapid and wide distribution in the tissues throughout the whole body within the time course examined. TCA precipitated radioactivity in kidneys was the highest, however, only trace amounts of [125I]exendin-4 was detected in the brain.

Urinary excretion of prostaglandin E2. (PGE2) and thromboxane B2 (TXB2, a stable metabolite of TXA2), atrial natriuretic peptide (ANP) and arginine vasopressin (AVP) in plasma were determined with radioimmunoassay in 30 healthy normal controls (NC) and 53 cirrhotic patients without asoites (n = 18), with ascites (n = 25) and with hepatorenal syndrome (HRS, n=10). All of four vasoactive substances in the group of cirrhotic patients without ascites were significantly higher than that in NG (P

Objective This study aimed to investigate the role of pyrroloquinoline quinone (PQQ) in repairing oxidative nerve cells,and to study the antioxidant capacity of PQQ on the oxidative damage of rats caused by apolexis,as well as the effects on learning and memory abilities of apolexis rats.Methods Oxidative damage of PC12 was induced by H2O2,and the repairing rate of PQQ on oxidative PC12 cells was tested by methylthiazolyldiphenyl-tetrazolium bromide assay kit.The 18-month-old male SD rats were administered PQQ (0,10,20,40 mg/kg).After 4 weeks,Morris water maze test was used to test the learning and memory ability.After 6 weeks,serum and brain tissue related indicators and antioxidant capacity were recorded.Results The survival rate of PC12 cells increased from 59.1％ to 90.5％ with 200 nmol/L PQQ.Compared with the apolexis model group,the latency of the PQQ group (20,40 mg/kg) was shortened in the Morris water maze experiment,the swimming distance was reduced,pass-through counts were increased,and the first secure platform pass-through was reduced.Meanwhile,the levels of malondialdehyde and lipofuscin in serum and brain tissue of PQQ group decreased,the levels of superoxide dismutase,glutathione peroxidase vitality,antioxidant capacity of PQQ group (20,40 mg/kg) were enhanced.Conclusion PQQ could repair the oxidative damage of nerve cells,and it was confirmed that PQQ could play the same antioxidant effect in body and brain,and increase the learning and memory ability of apolexis rats.

Objective To investigate the infection status of HCV in remunerated blood donors and risk factors in Jiangsu province.Methods A Cross-sectional study was conducted among people aged ＞50 years.Questionnaires were used to collect the information about their demographic characteristics and risk behaviors,and venous blood samples were collected from them to detect HCV anti-body,HCV-RNA and other biochemical indicators.EpiData and Stata were used for data entry and statistical analysis.Results The overall HCV sero-prevalence rates were 22.55％ and 61.05％ among remunerated blood donors.Data firom multiple stepwise regression analysis showed that alanine aminotransferase (ALT) (adjusted OR=1.38,95％CI:1.18-1.62) and aspartate aminotransferase (AST) (adjusted OR=l.30,95％CI:1.10-1.54) were associated with the outcomes of HCV infection,and fasting plasma glucose (adjusted OR=1.17,95％CI:1.01-1.35) were associated with HCV RNA viral loads.Conclusion The prevalence of HCV infection in remunerated blood donors was high,clinical ALT,AST and fasting plasma glucose levels were associated with the risk for HCV infection and HCV RNA viral load.

Objective:To explore the clinical and endoscopic features of patients with autoimmune gastritis (AIG) and to improve the accuracy of clinical diagnosis of AIG.Methods:From January 3, 2020 to November 25, 2021, the general information (gender, age), laboratory examination indicators and endoscopic findings of 179 AIG patients diagnosed at the Second Affiliated Hospital of Chongqing Medical University were retrospectively analyzed. The laboratory examination indicators included hemoglobin, gastrin-17, pepsinogen (PG), anemia combination indicators (ferritin, vitamin B 12), thyroid function indicators (thyroid-stimulating hormone, thyroglobulin antibody and thyroid peroxidase antibody), Helicobacter pylori, and anti-parietal cell antibody and anti-intrinsic factor antibody. Descriptive methods were used for statistical analysis. Results:Among the 179 AIG patients, there were 42 males (23.5%) and 137 females (76.5%), with an average age of (55.23±12.04) years old. The gastrin-17 level of AIG patients was 195.31 ng/L (143.64 ng/L, 273.61 ng/L), PG Ⅰ level and PG Ⅰ/PG Ⅱ ratio were 12.40 μg/L (7.65 μg/L, 19.40 μg/L) and 1.03 (0.66, 1.52), respectively. There were 15.3% (18/118) of the AIG patients with iron deficiency anemia, and 16.1% (19/118) with megaloblastic anemia. The positive rate of anti-parietal cell antibody was 71.8% (51/71), and the positive rate of anti-intrinsic factor antibody was 25.4% (18/71). The serum thyroid-stimulating hormone level increased in 27.3% (15/55) of the patients, and the positive rates of thyroglobulin antibody and thyroid peroxidase antibody were 31.6% (12/38) and 47.4% (18/38), respectively. The positive rate of Helicobacter pylori was 29.7% (38/128). The endoscopic appearance of AIG indicated reverse atrophy, characterized by obvious atrophy in gastric fundus and gastric body mucosa, however the atrophy of gastric antrum was not obvious. Under endoscopy yellow-white turbid mucus, which was difficult to be washed, was found in 67.0% (120/179) of the patients, and under endoscopy the residual gastric fundus glands could be seen in 19.6% (35/179) of the patients. Among 179 AIG patients, 7 cases (3.9%) of neuroendocrine tumor (NET), 7 cases (3.9%) of early gastric adenocarcinoma (including 1 case of poorly differentiated adenocarcinoma), 1 case (0.6%) of adenoma, and 14 cases (7.8%) of hyperplastic polyps were found. Except for the case of poorly differentiated adenocarcinoma undergoing surgery, the others were treated with endoscopic resection. Conclusions:When unexplained iron deficiency anemia, megaloblastic anemia, or reverse atrophy is found, AIG should be considered. AIG patients are at high risk for gastric cancer and NET, and should be closely followed up, and active treatment should be given before anemia and neurological symptoms appear.

Objective:To investigate the effect of reverse saphenous nerve neurocutaneous flaps for skin defects of forefoot with the help of the three dimensional computerized tomography angiography(3D-CTA).Methods:From November 2013 to February 2018, 19 patients with anterior foot defects were treated in the Yidu Central Hospital of Weifang. There were 12 males and 7 females. The patients ranged from 16 to 45 years old, with an average age of 29.6 years. Preoperative 3D-CTA examination of the ipsilateral foot was performed to find out the blood vessels in the donor site, to determine the location, course, length of the superficial branch of the medial plantar toe artery and its relationship with the surrounding tissues, and to design the medial saphenous nerve nutrient vessel skin flap for retrograde repair of the skin and soft tissue defect at the front of the foot. Sensitive recovery were evaluated according to Swanson evaluation of clinical effect of peripheral nervous impairment.Results:Preoperative three-dimensional CT angiography showed that the superficial branches of the medial plantar toe artery were detected, and the perforating vessels during the operation were basically consistent according to the result of the examination. The 19 cases of forefeet skin detects were successfully reconstructed with the reverse saphenous nerve neurocutaneous flaps. All patients were followed up with the mean of 8 months. All flaps survived totally without diabrosis and swelling. The walking and weight-bearing were normal and the blood supply of foot were good. The functional recovery of foot and ankle was excellent in 8 cases, good in 10 cases and middle in 1 case. The sensory recovery of the forefoot: S4 5 feet, S3 10 feet, S2 4 feet; R4 3 feet, R3 9 feet, R2 7 feet.Conclusions:Preoperative 3D-CTA examination can confirm the anatomy of the superficial branch of the medial toe base artery in the donor site and guide the design of the medial saphenous nerve nutrient vessel flap for retrograde repair of skin and soft tissue defects of the distal foot. The blood supply is reliable, the wound is small and the effect is satisfactory.

Objective:To establish a new model for the prediction of severe outcomes of COVID-19 patients and provide more comprehensive, accurate and timely indicators for the early identification of severe COVID-19 patients.Methods:Based on the patients’ admission detection indicators, mild or severe status of COVID-19, and dynamic changes in admission indicators (the differences between indicators of two measurements) and other input variables, XGBoost method was applied to establish a prediction model to evaluate the risk of severe outcomes of the COVID-19 patients after admission. Follow up was done for the selected patients from admission to discharge, and their outcomes were observed to evaluate the predicted results of this model.Results:In the training set of 100 COVID-19 patients, six predictors with higher scores were screened and a prediction model was established. The high-risk range of the predictor variables was calculated as: blood oxygen saturation <94 %, peripheral white blood cells count >8.0×10 9, change in systolic blood pressure <-2.5 mmHg, heart rate >90 beats/min, multiple small patchy shadows, age >30 years, and change in heart rate <12.5 beats/min. The prediction sensitivity of the model based on the training set was 61.7 %, and the missed diagnosis rate was 38.3 %. The prediction sensitivity of the model based on the test set was 75.0 %, and the missed diagnosis rate was 25.0 %. Conclusions:Compared with the traditional prediction (i.e. using indicators from the first test at admission and the critical admission conditions to assess whether patients are in mild or severe status), the new model’s prediction additionally takes into account of the baseline physiological indicators and dynamic changes of COVID-19 patients, so it can predict the risk of severe outcomes in COVID-19 patients more comprehensively and accurately to reduce the missed diagnosis of severe COVID-19.

Objective:To investigate the effect of reverse saphenous nerve neurocutaneous flaps for skin defects of forefoot with the help of the three dimensional computerized tomography angiography(3D-CTA).Methods:From November 2013 to February 2018, 19 patients with anterior foot defects were treated in the Yidu Central Hospital of Weifang. There were 12 males and 7 females. The patients ranged from 16 to 45 years old, with an average age of 29.6 years. Preoperative 3D-CTA examination of the ipsilateral foot was performed to find out the blood vessels in the donor site, to determine the location, course, length of the superficial branch of the medial plantar toe artery and its relationship with the surrounding tissues, and to design the medial saphenous nerve nutrient vessel skin flap for retrograde repair of the skin and soft tissue defect at the front of the foot. Sensitive recovery were evaluated according to Swanson evaluation of clinical effect of peripheral nervous impairment.Results:Preoperative three-dimensional CT angiography showed that the superficial branches of the medial plantar toe artery were detected, and the perforating vessels during the operation were basically consistent according to the result of the examination. The 19 cases of forefeet skin detects were successfully reconstructed with the reverse saphenous nerve neurocutaneous flaps. All patients were followed up with the mean of 8 months. All flaps survived totally without diabrosis and swelling. The walking and weight-bearing were normal and the blood supply of foot were good. The functional recovery of foot and ankle was excellent in 8 cases, good in 10 cases and middle in 1 case. The sensory recovery of the forefoot: S4 5 feet, S3 10 feet, S2 4 feet; R4 3 feet, R3 9 feet, R2 7 feet.Conclusions:Preoperative 3D-CTA examination can confirm the anatomy of the superficial branch of the medial toe base artery in the donor site and guide the design of the medial saphenous nerve nutrient vessel flap for retrograde repair of skin and soft tissue defects of the distal foot. The blood supply is reliable, the wound is small and the effect is satisfactory.