OBJECTIVE: To investigate the effects of calcitonin gene-related peptide (CGRP) on autophagy in hypoxic/reoxygenated (H/R) cardiomyocytes and its relationship with the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathway. METHODS: The rat cardiomyocyte cell line H9c2 was routinely cultured in vitro and passaged for experiments when the cells grew to 80% fusion. (1) CGRP dosage screening experiment: the cells were divided into blank control group, H/R group and different dosages of CGRP pretreatment groups. H9c2 cells were placed in a closed hypoxia chamber for 2 hours and then reoxygenated in a conventional incubator for 12 hours to prepare the H/R model. The CGRP pretreatment groups were pretreated with 0.01, 0.1, 0.5, 1, 5, and 10 μmol/L CGRP before the modeling process. The blank control group was not given any treatment. Cell counting kit-8 (CCK-8) was used to detect the cell survival rate, and the most suitable drug dosage was screened out. (2) Intervention experiment: H9c2 cells were divided into blank control group, H/R group, CGRP+H/R group, and CGRP+PI3K target inhibitor ly294002 (LY)+H/R group. H/R group was prepared as cellular H/R model. CGRP (1 μmol/L) alone or in combination with LY (10 μmol/L) was administered to CGRP+H/R group and CGRP+LY+H/R group, respectively, prior to the preparation of cellular H/R model. The blank control group was cultured routinely without treatment. The cell survival rate was detected by CCK-8. The level of lactate dehydrogenase (LDH) release was detected by colorimetric assay. The expressions of autophagy-related proteins [autophagy effector protein Beclin-1, microtubule-associated protein 1 light chain 3-II (LC3-II), autophagy protein p62] and PI3K/Akt/mTOR signaling pathway proteins [phosphorylated Akt (p-Akt), phosphorylated mTOR (p-mTOR)] were detected by Western blotting. RESULTS: (1) Results of CGRP dosage screening experiment: compared with the blank control group, the cell survival rate of the H/R group decreased significantly; and after giving 0.1, 0.5, 1, 5 μmol/L CGRP for pretreatment, the cell survival rate increased significantly, and intervention effect of 1 μmol/L CGRP was the best, and the difference was statistically significant when compared with that of the H/R group [(74.23±6.18)% vs. (23.43±4.09)%, P < 0.01], so it was used as the intervention dosage for the subsequent experiment. (2) Intervention experiment results: compared with the blank control group, the cell survival rate in the H/R group was significantly reduced, the level of LDH release was significantly increased, the protein expressions of Beclin-1 and LC3-II were significantly increased, and the protein expressions of p62, p-Akt and p-mTOR were significantly reduced, indicating that the death of cardiomyocytes occurred after the treatment of H/R and was accompanied by the elevation of autophagy level, and this process was associated with the activation of PI3K/Akt/mTOR signaling pathway. Compared with the H/R group, CGRP pretreatment increased cell survival rate [(76.02±2.43)% vs. (46.15±3.29)%, P < 0.01], decreased the level of LDH release (U/L: 169.83±11.65 vs. 590.17±34.50, P < 0.01), and down-regulated the protein expressions of Beclin-1 and LC3-II [Beclin-1 protein (Beclin-1/β-actin): 1.27±0.15 vs. 1.93±0.19, LC3-II protein (LC3-II/LC3-I): 1.27±0.13 vs. 1.98±0.18, both P < 0.01], up-regulated the protein expressions of p62, p-Akt, p-mTOR [p62 protein (p62/β-actin): 0.96±0.02 vs. 0.63±0.05, p-Akt protein (p-Akt/Akt): 0.76±0.04 vs. 0.48±0.02, p-mTOR protein (p-mTOR/mTOR): 1.13±0.09 vs. 0.68±0.15, all P < 0.05], suggesting that CGRP was able to reduce the H/R-induced cardiomyocyte injury, and this process was accompanied by a decrease in the level of cellular autophagy and activation of the PI3K/Akt/mTOR signaling pathway. Compared with the CGRP+H/R group, the cell survival rate was significantly lower than that in the CGRP+LY+H/R group [(56.95±6.63)% vs. (76.02±2.43)%, P < 0.01], LDH release level was significantly higher (U/L: 436.00±27.44 vs. 169.83±11.65, P < 0.01), and the protein expressions of Beclin-1 and LC3-II were significantly up-regulated [Beclin-1 protein (Beclin-1/β-actin): 1.63±0.12 vs. 1.27±0.15, LC3-II protein (LC3-II/LC3-I): 1.61±0.13 vs. 1.27±0.13, both P < 0.01], and significantly down-regulated p62, p-Akt, and p-mTOR protein expressions [p62 protein (p62/β-actin): 0.57±0.09 vs. 0.96±0.02, p-Akt protein (p-Akt/Akt): 0.45±0.01 vs. 0.76±0.04, p-mTOR protein (p-mTOR/mTOR): 0.66±0.06 vs. 1.13±0.09, all P < 0.05], suggesting that PI3K-targeted inhibitor was able to reverse the protective effect of CGRP on H/R cells. CONCLUSIONS CGRP pretreatment attenuated H/R-induced cardiomyocyte injury, increased cell survival rate, and reduced cellular LDH release. This effect may be achieved through inhibiting the activation of PI3K/Akt/mTOR signaling pathway.
Animals ; Myocytes, Cardiac/drug effects* ; Signal Transduction/drug effects* ; Rats ; TOR Serine-Threonine Kinases/metabolism* ; Calcitonin Gene-Related Peptide/pharmacology* ; Proto-Oncogene Proteins c-akt/metabolism* ; Autophagy/drug effects* ; Cell Line ; Cell Hypoxia ; Phosphatidylinositol 3-Kinases/metabolism*

Objective:To study the changes in long non-coding RNA C2dat1 expression in kidney tissues of rats at different stages of diabetic kidney disease (DKD) and its relationship with renal interstitial fibrosis.Methods:Forty-eight male SD rats were randomly divided into two groups with 24 rats in each group: control group and DKD group. The rats in the control group were fed with ordinary diet, while those in the DKD group were fed with high-fat diet and drank water freely. After eight weeks of feeding, the rats were fasted for 12 h with free access to water. Then, the DKD group was given a one-time intrabitoneal injection of streptozotocin and the control group was given an equal dose of sodium citrate buffer. After 72 h, the random peripheral blood glucose concentration (≥ 16.7 mmol/L for three consecutive days) and urine sugar (positive) were tested to assess the establishment of the diabetes model. Urine, blood and kidney samples were collected at 3, 6, 9 and 12 weeks. The urinary protein excretion rate within 24 h, urinary creatinine and serum total cholesterol were measured by automatic biochemical apparatus. Pathological changes in kidney tissues were observed by HE staining. The expression of calcium/calmodulin-dependent protein kinase Ⅱ delta (CaMK2D), p65, p50, α-SMA and E-cardherin was detected by immunohistochemistry. Quantitative real-time PCR (qPCR) was used to detect the expression of lncRNA C2dat1 and CaMK2D. The relationship of lncRNA C2dat1 with α-SMA, E-cardherin and CaMK2D was analyzed by correlation analysis. In in vitro experiment, renal tubular epithelial cells HK-2 were induced by high glucose. The expression of lncRNA C2dat1 and CaMK2D in HK-2 cells was detected by qPCR after 24, 48 and 72 h of intervention. Results:The rats in the DKD group showed typical symptoms such as polydipsia, polyphagia, significant weight loss and increased blood glucose as compared with the rats in the control group. Results of the biochemical tests revealed that compared with the control group, the DKD group had increased 24 h excretion rate of urinary protein, decreased urinary creatinine and up-regulated total cholesterol. HE staining showed that the rats in the control group had intact glomeruli, normal basement membrane and no mesangial hyperplasia or inflammatory cell infiltration. However, enlarged glomeruli and evenly thickened basement membrane were observed in the DKD group. Immunohistochemistry indicated that the expression of CaMK2D, p50 and α-SMA was higher in the DKD group than in the control group, while the expression of E-cardherin was lower in the DKD group. qPCR results showed that the expression of lncRNA C2dat1 and CaMK2D at mRNA level was higher in the DKD group than in the control group. In in vitro experiment, the expression of lncRNA C2dat1 and CaMK2D at mRNA level was also higher in HK-2 cells induced by high glucose than in the control group. Correlation analysis indicated that lncRNA C2dat1 was positively correlated with α-SMA and CaMK2D, but negatively correlated with E-cardherin. Conclusions:During the progression of DKD, the high expression of lncRNA C2dat1 might promote diabetic renal interstitial fibrosis by regulating the expression of CaMK2D to activate the NF-κB signaling pathway.

Objective:To explore the features of free uroflow(FF) curve patterns in female patients with detrusor underactivity(DU) and their clinical significance.Methods:Data of 275 adult female patients with lower urinary tract symptoms(LUTS) underwent urodynamic studies(UDS) at urology center of our hospital from June 2014 to June 2016 were analyzed retrospectively. The uroflow curve patterns of patients with DU were classified and analyzed in the context of parameters of FF, cystometry (CM), and pressure-flow study(PFS). The prevalence of each abnormal uroflow curve pattern in DU patients were calculated and compared with those in non-DU patients.Results:No bell-shaped curve was found in 141 patients with DU. The abnormal curve patterns can be divided into 5 types: Type Ⅰ (bell-shaped curve with saw tooth) in 20 cases (14.2%), Type Ⅱ (box-like curve) in 34 cases (24.1%), Type Ⅲ (triangle curve with decreasing slop) in 62 cases(43.9%), Type Ⅳ (triangle curve with increasing slop) in 4 cases (4.3%), Type Ⅴ (tide-wave curve)in 19 cases (13.5%). Maximum flow rate of free uroflow(Q max.FF) of type Ⅰ [(28.4±9.7) ml/s] was significantly greater than that of type Ⅱ, Ⅲ and Ⅴ[(17.0±4.1), (15.8±5.4) and (12.9±6.4) ml/s, P<0.05]. Flow time of free uroflow(FT.FF) of type Ⅲ and Ⅴ [(43.7±17.2) and (50.1±28.9)s] were significantly longer than that of type Ⅰ and Ⅱ [(18.5±7.3)s and (27.2±9.7)s, P<0.05]. Post voided residual > 50ml was noted in 19 cases (30.6%) of type Ⅲ, 7 cases (36.8%) of type Ⅴ, 1 case (2.9%) of type Ⅱ and no one in type Ⅰ and Ⅳ. Abnormal manifestations in cystometry mainly included bladder hypersensitivity, detrusor overactivity, and stress urinary incontinence. Detrusor pressure at Q max (Pdet.Q max) of type Ⅴ [(7.4±5.0) cmH 2O] was significantly lower than that of type Ⅰ, Ⅱ, Ⅲ [(11.8±6.7), (12.0±5.3), (12.1±5.0) cmH 2O, P<0.05]. Among 134 cases of non-DU, there were type Ⅰ curves in 88 cases (65.7%), type Ⅱ curves in 4 cases (2.9%), type Ⅲ curves in 15 cases (11.2%), type Ⅳ curves in 1 cases (0.7%), type Ⅴ curves in 7 cases (5.2%). And normal bell-shaped curves in 19 cases(14.2%). The prevalence of type Ⅱ, Ⅲ and Ⅴ in DU patients was significantly higher than that in the non DU patients ( P<0.05). Conclusions:This study reveals that the characteristics of reduced detrusor contractility and duration, prolonged bladder emptying or incomplete emptying can be reflected in the patterns of free uroflow curve in female patients with DU. The abnormalities of these free uroflow curve patterns, especially type Ⅱ, Ⅲ and Ⅴ will be helpful in preliminarily screening DU in females.

Objective:To explore the efficiency of a new scoring system of gastric cancer screening for early gastric cancer in health examination population.Methods:The risk score of gastric cancer was assessed based on the new scoring system in health examination population. A notice for further gastroscopy was sent to the medium-risk and high-risk people. Gastroscopy was performed on those who agreed to undergo the examination.Results:From January to April 2019, a total of 5 357 people in health system visited the Physical Examination Center of Shanghai Songjiang Clinical Medical College of Nanjing Medical University for health examination. Seven hundred and forty people were classified as medium- and high-risk groups by the new screening system, 576 in medium-risk group, and 164 in high-risk group. Among them, 131 cases (17.70%) came for further gastroscopy, of whom 91 (69.47%) were in the medium-risk group and 40 (30.53%) in the high-risk group. After gastroscopy, 4 cases of gastric cancer and 1 case of esophageal cancer were detected, and both were early cancer. In the medium-risk group, 2 cases (2/91, 2.20%) of early gastric cancer and 1 case (1/91, 1.10%) of early esophageal cancer were found. In the high-risk group, 2 cases (2/40, 5.00%)of early gastric cancer were found. The tumor detection rate of high-risk group (5.00%) was higher than that of medium-risk group (3.30%), but there was no significant difference ( P>0.05). Conclusion:Risk stratification with the new scoring system of gastric cancer screening can improve the detection rate of early gastric cancer.

OBJECTIVE: To investigate the correlation between the severity of obstructive sleep apnea syndrome (OSAS) and red cell distribution width (RDW) in elderly patients. METHODS: A cross-sectional study was conducted among 311 elderly patients diagnosed with OSAS in the snoring clinic between January, 2015 and October, 2016 and 120 healthy controls without OSAS from physical examination populations in the General Hospital of PLA. The subjects were divided into control group with apnea-hypopnea index (AHI) <5 (=120), mild OSAS group (AHI of 5.0-14.9; =90), moderate OSAS group (AHI of 15.0-29.9; =113) and severe OSAS group (AHI ≥ 30; =108). The clinical characteristics and the results of polysomnography, routine blood tests and biochemical tests of the subjects were collected. Multiple linear regression analysis was used to examine the correlation between OSAS severity and RDW. RESULTS: The levels of RDW and triglyceride were significantly higher in severe OSAS group than in the other groups ( < 0.01). The levels of fasting blood glucose and body mass index were significantly higher in severe and moderate OSAS groups than in mild OSAS group and control group ( < 0.05 or < 0.01). Multiple linear regression analysis showed that AHI was positively correlated with body mass index (β=0.111, =0.032) and RDW (β=0.106, =0.029). The area under ROC curve of RDW for predicting the severity of OSAS was 0.687 (=0.0001). CONCLUSIONS The RDW increases as OSAS worsens and may serve as a potential marker for evaluating the severity of OSAS.
Aged ; Cross-Sectional Studies ; Erythrocyte Indices ; Humans ; Polysomnography ; Severity of Illness Index ; Sleep Apnea, Obstructive

Objective:To explore the anti-epidemic preventions and perioperative management strategies of organ donation and liver transplantation during the pandemic period of novel coronavirus pneumonia (NCP) and summarize the experiences.Methods:On the basis of guidance of National Health Commission and Organ Transplantation Committee of Chinese Medical Association, anti-epidemic preventions and perioperative management strategies of organ donation and liver transplantation were adjusted under the background of NCP pandemic and the anti-epidemic preventions and treatment outcomes were evaluated. Eight organ donations and 7 liver transplantations were performed from February 4 to March 7, 2020. NCP infection screening results were negative in all pre-donation and pre-transplantation cases.Results:All donation operations and liver transplantations were successfully performed without postoperative complications. No NCP occurred during hospitalization period. Postoperative pulmonary infection occurred in 1 case (1/7) and the following novel coronavirus screening result was negative. Pulmonary inflammation became partially absorbed after antibacterial therapy.Conclusions:Through strict and effective anti-epidemic preventions and perioperative managements, organ donation and transplantation could be successfully performed during the pandemic period of NCP.

Objective:To explore the morphology and vessel distribution of the scapholunate interosseous ligament and anatomical basis for the clinical reconstruction of scapholunate interosseous ligament.Methods:From October, 2018 to December, 2018, 12 fresh wrist joint specimens were perfused with gelatin-lead oxide solution from ulnar or radial artery and scanned under micro-CT. The morphology of scapholunate interosseous ligament in neutral position and the distribution of nutrient vessels in the ligament were observed on reconstructed 3D images by Mimics. The width, length and thickness of palmar, dorsal and proximal ligaments were measured. The anatomical parameters at the entrance of nutrient vessels in the scapholunate interosseous ligament were taken and their relationship with the blood supply to the scapholunate was analyzed.Results:①For scapholunate interosseous ligament, it was found that the average length of the proximal sub-region was the longest, the length of palmar and dorsal sides was similar to each other and the widest and thinnest was in palmar side, while the thickness and width of dorsal and proximal were similar. ②There was no nutrient vessel in the proximal part of the scapholunate interosseous ligament. But there were abundant nutrient vessels in the palmar and dorsal scapholunate interosseous ligament, and there was no significant difference in blood supply to palmar and dorsal scapholunate interosseous ligament ( P>0.05). ③The palmar and dorsal medial nutrient vessels that supply to the scapholunate interosseous ligament enter the scapholunate from the attachment of ligament of scapholunate interosseous joint. Conclusion:The palmar side of the scapholunate interosseous ligament is wider and thinner than that of the other subareas, which makes it more vulnerable to injury from an anatomical point of view. There is abundant blood supply to the palmar and dorsal subareas of the scapholunate interosseous ligament and the supplying vessels anastomose inside the scapholunate bone. There is no distribution of blood vessel at the proximal part of scapholunate interosseous ligament, hence is difficult to heal. An injury of palmar and dorsal ligaments may affect the blood supply of scapholunate.

Objective This study aimed to investigate the renal expression change of high mobility of nucleosome binding protein 1 ( HMGN1) in epithelia-mesenchymal transition ( EMT) process, and to study the effect of HMGN1 on renal fibrosis in the diabetic nephropathy mice model. Methods 20 C57BL/6 mice were randomly divided into control group, model group, benazepril group, and insulin group. After 8 weeks of drug intervention, blood, urine and kidney tissue samples were taken from mice. The routine physiological and biochemical indexes were detected. Renal structure and fibrosis were detected by HE and Sirius red staining, respectively. Immunohistochemistry and in situ hybridization were used to investigate the protein and mRNA expression levels of HMGN1, CD68, F4/80,α-smooth muscle actin (α-SMA) , and E-cadherin in renal tissue. Results Blood glucose, renal index, and urinary albumin to creatinine ratio ( UACR) were significantly higher in the model group than those in the normal group. In the model group, HE staining showed glomerular hypertrophy and interstitial inflammatory cell infiltration, and Sirius red showed collagen deposition in the renal tissue. Compared with normal group, HMGN1, CD68, F4/80 positive cell counts andα-SMA protein expression were all increased, while E-cadherin protein expression was downregulated in the model group ( all P<0.05) . The above indexes were not improved significantly in the benazepril group. And after intervention of insulin, the expression levels of CD68 positive cell count andα-SMA protein were decreased and the expression levels of E-cadherin protein were increased compared with the model group ( all P<0.05) . The correlation analysis showed that the level of HMGN1 was correlated with CD68, F4/80, α-SMA, E-cadherin and collagen protein, while CD68 and f4/80 were correlated withα-SMA, collagen protein and blood glucose, respectively ( all P<0. 05 ) . Conclusion HMGN1 is involved in the progression of diabetic nephropathy fibrosis, and its underlying mechanism might be related to the macrophage-mediated EMT process.

Objective To approach the spine MRI features and its possibility of etiology for refractory lower urinary tract symptoms(LUTS) in female patients.Methods We conducted prospectively a cross sectional description study of female patients with refractory LUTS during January 16 through March 27 in 2017 based on a urologist's outpatient work.The including criteria were adult female patients with refractory LUTS which defined as having LUTS more than three months and having poor response to behavior therapy and medication treatment.The excluding criteria were patients having evidence of infection,tumor,stone in urinary tract,any central nerve system diseases,or any other diseases may potentially producing LUTS.Data collected included patients demographic information,main complains,present disease features,disease history,physical examination,urine routine,urodynamic study and spine MRI.The characteristics of clinical manifestation,urodynamic study and spine MRI were analyzed.Results During the time span of study,totally 70 cases had been diagnosed as having refractory LUTS and had qualified data of clinical recordings,urodynamic study and spine MRI.Among these 70 cases,63 (90.9％) had storage phase symptoms,11 (15.7％) had voiding phase symptoms,8 (11.4％) had postmicturition symptoms,12 (17.1％) also had disorders in defecating,45 (64.3％) had pain in lower abdomen or pelvic region.69 cases (98.6％) had urodynamic disorders,33 (47.1％) had oversensitivity of bladder,12 (17.1％) had smaller bladder volume,16(22.9％) had detrusor overactivity,15 (21.4％) had bladder outlet obstruction,39(55.7％) had detrusor underactivity.69 cases(98.6％)had spine MRI abnormalities,54(77.1％) had sacral nerve lesions,49 (70.0％) had cervical lesions,48 (68.6％) had lumbar lesions,4 had thorathic lesions.Conclusions The present study revealed extraordinary high prevalence of abnormality in urodynamic parameters and spine MRI in female patients with refractory LUTS,which implies possibility that the refractory LUTS are caused by lesions in spinal nerve system.

Objective To investigate the effect of subclinical epileptiform discharges (SED) on the cognition of adult patients with epilepsy,exploring the mechanism of SED that leads to cognitive impairment in adult patients with epilepsy to raise physicians' attention about SED.Methods Patients were collected in the Department of Neurology,China-Japan Union Hospital of Jilin University from March 2016 to March 2017.Sixty adult patients with epilepsy without clinical episodes in the last three months were selected as SED group and 40 healthy volunteers as control group.Medical history of the SED group was recorded in detail.All patients were examined by Self-rating Anxiety Scale,Self-rating Depression Scale,Pittsburgh Sleep Quality Index scale in order to exclude organic brain disorders,metabolic diseases,anxiety,depression,sleep disorders and drug-induced cognitive dysfunction.Subjects in the two groups received Montreal Cognitive Assessment (MoCA),electroencephalogram and blood oxygen level dependent functional magnetic resonance imaging examination.Finally,the results were compared between the two groups.Results ①SED had different effects on cognitive function in adult patients with epilepsy,and the MoCA score (26(22,27)) showed statistically significant difference compared with the control group (29 (28,29),Z =-6.26,P =0.00).②Different discharges indexes showed different effects on cognitive function aspects.Cognitive impairment was significant when the discharges indexes were ＞ 10％ (discharges indexes 1％-10％:MoCA score 26(26,28),discharges indexes 10％-50％:MoCA score 22(19.5,25),Z =-4.74,P =0.00).③The cognitive function of epilepsy patients was positively correlated with the duration of education (r =0.41,P =0.00) and the time interval to recent seizure (r =0.31,P =0.02),and negatively correlated with SED (r =-0.57,P=0.17).There was no correlation between cognitive function and duration of disease and onset age.The SED was the main influencing factor of cognitive function in epilepsy.④Compared with healthy people,epilepsy patients with SED showed differences in resting brain function network connection,with strong connective regions at the right inferior temporal gyrus,right hippocampus,bilateral thalamus,with weak connective regions at the double medial upper frontal gyrus,lateral dorsal frontal gyurs.Conclusions SED had an effect on the cognitive function of adult patients with epilepsy.The mechanism of cognitive impairment in adult epilepsy with SED may be related to abnormal brain function in cognitive-related areas.