Objective To study the chemical constituents in the immature fruits of Momordica charantia. Methods Isolation and purification were carried out by macroporous absorption resin and silica gel, and compounds were identified and elucidated by spectral and chemical methods. Results Ten compounds were obtained from alcohol extract and five of them were determined as germanicyl acetate (Ⅰ), aglycone of momordicoside Ⅰ (Ⅱ), aglycone of momordicoside L (Ⅲ), charantin (Ⅳ), and ?-sitosterol (Ⅴ). Conclusion For all the first time, compound Ⅰ is found in the plants of Momordica L., compound Ⅱ is found as novel natural product in this plant, and compound Ⅲ is found in China.

Objective To study the chemical constituents in Momordica charantia.Methods The alcohol extract was isolated by macroporous adsorption resin and silica gel,and the compound structures were identified by spectral methods.Results Two couple isomers were isolated,the structure of one couple was identified as 19R-5?,19-epoxycucurbita-6,23,25-trien-3?,19-diol(Ⅰa)and 19S-5?,19-epoxycucurbita-6,23,25-trien-3?,19-diol(Ⅰb);the structure of the other couple was identified as 3-O-glucopyranoside of 5?,19-epoxycucurbita-6,23,25-trien-3-ol(Ⅱa),and 3-O-allopyranoside of 5?,19-epoxycucurbita-6,23,25-trien-3-ol(Ⅱb).Conclusion New compounds Ⅰa,Ⅰb,Ⅱa,and Ⅱb are isolated from M.charantia for the first time.

Objective:To observe the bone marrow mesenchymal stem cell isolation,identification methods and protection mechanisms in the kidney of diabetic rats.Methods: Thirty-one rats were used to isolate bone marrow mesenchymal stem cells (BMSCs) by adherent culture method.Cell morphology was observed by phase contrast microscope.The surface molecular weight of cultured cells was detected by Real-time PCR.30 rats were induced by intraperitoneal injection of streptozotocin to establish diabetic rat model (n=15).And stem cell treatment group (n=15) according to the different treatment methods.Rats in the control group were treated with insulin combined with probucol.Stem cell treatment group was implanted with stem cells.Before and after treatment,fasting blood glucose,24 h urinary protein excretion,creatinine clearance rate and kidney weight were measured.PAS staining.And the renal tissue was detected by Western blot.Results: The primary bone marrow mesenchymal stem cells (MSCs) were cultured for 24 hours,and the cells were irregularly distributed in the culture flask.After 7 days of culture,the cells were regular,elongated,elliptical,and strong refraction,and the cells were fused with each other.The morphology of BMSCs was uniform and showed a long fusiform shape,CD44,CD31 and CD34 were observed by RT-PCR.The levels of fasting blood glucose,24 h urine protein excretion and creatinine clearance rate in the observation group were significantly higher than those in the control group (P<0.05).The results of PAS staining showed that the glomerular structure was clear,the morphology was regular,the arrangement rules of glomerular cells,the basement membrane and the encapsulation of the glomerular membrane were significantly higher than those of the control group Man′s capsule clear (P<0.05).In the control group,the glomerular volume increased,the mesangial area widened and the stroma increased significantly.The Bcl-2 protein level and the Bax and Caspase-3 protein levels were significantly decreased in the stem cell treatment group.The levels of Bcl-2 and Bax and Caspase-3 protein in the control group were significantly decreased (P<0.05).The ROS,MDA and SOD levels in oxidative stress group were lower than those in control group (P<0.05).The difference of ROS,MDA and SOD between the two groups was not statistically(P>0.05).Conclusion: It is suggested that BMSCs can be isolated from rat bone marrow mesenchymal stem cells by using adherent culture method and labeled with green fluorescent protein in vitro.Bone marrow mesenchymal stem cells can protect the kidney and inhibit the oxidative stress of diabetic kidney.

Bronchopulmonary dysplasia(BPD)is one of the most serious lung diseases in premature infants and an important cause of death in preterm infants.The pathophysiological mechanism of BPD is still unclear.The preventive and targeted treatment strategies for this disease are also very limited.However, various biomarkers have been found in blood, alveolar lavage fluid, exhaled gas condensate, and urine.It is used to identify high-risk children with BPD early and to predict the severity and prognosis of the disease.

Objective To investigate the appropriate waist circumference (WC) cutoff points for central obesity in the middle-aged and elderly Beijing residents by the metabolic syndrome definition of the International Diabetes Federation (IDF). Methods A total of 2,344Beijing residents aged ≥40 years were investigated. They answered questionnaires, received physical examinations, and underwent plasma glucose and lipid profile measurement. Those non-diabetic subjects underwent a 75g oral glucose tolerance test. All data were analyzed to calculate the appropriate WC cutoff points for central obesity reaching the diagonsis of MS. Results 1) Both in males and females, the triglyceride (TG), systolic blood pressure, diastolic blood pressure and fasting plasma glucose (FPG) increased linearly with WC, and the high density lipoprotein cholesterol (HDL-C) decreased linearly with WC (P＜0.05). 2)The prevalence of elevated TG,reduced HDL-C, elevated blood pressure, elevated FBG, or ≥ 2 of these factors increased with WC (P＜0.05). 3) Based on the receiver operating characteristic (ROC) curve analysis and Youden index, the WC values for central obesity and for detecting BMI ≥ 25 kg/m2were about 90 cm for men and 80 cm for women. 4) The odds ratio for the presence of two or more metabolic risk factors increased abruptly in men with WC ≥ 90 cm and in women with WC ≥ 80 cm. Conclusions The appropriate WC cutoff point for central obesity was determined to be 90 cm for men and 80 cm for women in the middle-aged and elderly Beijing residents by the metabolic syndrome definition of IDF.

Objective To investigate the molecular mechanism of solanine-induced apoptosis of prostate cancer cells Du145 and LNCaP.Methods The effects of solanine on the viability of Du145 and LNCaP cells were evaluated by MTT assay.The generation of intracellular reactive oxygen species (ROS) and solanine-induced apoptosis were measured by flow cytometry.The protein levels of p38 and p-p38 expressions were examined by Western blot.Results Solanine significantly inhibited the viability of Du145 and LNCaP cells in a dose-dependent manner (P＜0.01).The inhibition of solanine on cell viability was suppressed by the ROS scavenger NAC.ROS generation,apoptosis and phosphorylation of p38 were induced by treatment with solanine at 40 μmol/L for 24 h.The expression of p38 and solanine-induced apoptosis were suppressed by NAC and SB203580.Conclusion Solanine induces the apoptosis of human prostate cancer cell via the RO.S-p38 signaling pathway.

Objective To investigate the outcome and related risk factors of integrated intensive intervention in participants with impaired glucose regulation (IGR) after two years by the criteria of American Diabetes Association 2003. Methods The subjects who remained to be IGR at the end of first year following 75 g oral glucose tolerance test were randomly assigned to either a routine care control group or to an intensive integrated intervention group. The control group received general dietary and exercise advice at baseline and was followed up. In addition to dietary control and exercise advice, mefformin or acarbose were administrated in the intervention group. The latter group was also advised to take antihypertensive agents, lipid-regulating agents if necessary, as well as aspirin. Results The proportion of patients who fulfilled the assigned goals of blood glucose, blood pressure, body mass index or triglycerides was significantly higher in the intensive group than those in the control group. None in the intensive group developed overt diabetes mellitus, while 8 (9.3%) in the control group did. The proportion of patients who reverted to normal glucose tolerance (NGT) was slightly higher in the intensive group than in the control group (29.5% vs 22.1%, P>0.05). Logistic analysis showed that increase of waist circumference and systolic blood pressure was positively while the improvement of islet β-cell function was negatively correlated with the development of diabetes mellitus. Conclusions The intensive integrated intervention could significantly decrease the conversion rate of IGR to diabetes mellitus, and increase the chance of reversion to NGT. The increase of waist circumference or systolic blood pressure, the deterioration of islet β-ccll function were the influencing factors of the conversion of IGR to diabetes mellitus.

Objective:To investigate the serum 25-hydroxyvitamin D [25(OH) D] levels in elderly patients with hip and vertebral compression fractures (VCF).Method:Ninety patients (58 males and 32 females) aged over 60 years with hip fracture and 120 patients (88 males and 32 females) aged over 60 years with VCF admitted in the Aviation General Hospital from January 2017 to June 2019 were enrolled. Serum 25(OH)D levels were measured.Results:Serum level of 25 (OH) D in hip fracture patients was (9.0±6.8) μg/L, the 25 (OH) D level was lower than the normal value(<19.0 μg/L)in 79 patients and<3 μg/L in 24 patients. The level of 25(OH)D in VCF patients was (16.7±10.6) μg/L, the 25 (OH) D level was<19.0 μg/L in 78 patients (65.0%) and <3 μg/L in 10 patients (8.3%). The low level of 25(OH)D was negatively correlated with age in two groups ( r=-0.367, P=0.01; r=-0.313, P=0.04). The mean level of 25 (OH) D in the hip fracture group was lower than that in the VCF group ( t=5.960, P<0.01), and the low 25(OH)D rate in the former group was significantly higher than that in the latter group (χ 2=14.14, P<0.01; χ 2=12.74, P<0.01). The 25(OH)D value of female VCF patients was (14.5±8.8) μg/L, which was significantly lower than that of male patients (22.5±12.9) μg/L ( t=3.882, P<0.01).Among hip fracture patients, the 25(OH)D level in patients with fracture history was (8.3±6.9) μg/L, which was significantly lower than that of patients without fracture history (10.8±6.9) μg/L, and the difference was statistically significamt ( t=2.123, P=0.04). The serum osteocalcin level was (20.5±19.8) μg/L in patients with fracture history, which was significantly higher than that in patients without fracture history [(10.6±5.4) μg/L, t=3.245, P<0.01]. Conclusion:Elderly patients with new hip fractures have more severely low vitamin D level than patients with new VCF, and patients with previous fracture history have lower vitamin D levels than patients without fracture history.

Objective:To investigate the role of microRNA (miR)-497 in the formation of corneal neovascularization (CNV) induced by alkali burn and its mechanism.Methods:Forty-two wild type (WT) C57BL/6 mice aged 6 to 8 weeks, 42 CRISPR/Cas9 mediated miR-497 knockout (KO) and 42 CRISPR/Cas9 mediated overexpression transgenic (TG) C57BL/6 mice were selected and assigned as WT group, KO group and TG group, respectively.The corneal alkali burn model was established.At 3, 7, 14 and 21 days after modeling, corneal epithelium damage and stromal turbidity were scored according to slit lamp microscopy.The area of neovascularization was measured.Corneal structural changes and expression of inflammatory cells were observed by histopathological staining.The expression of CD31 in corneal tissues was detected by immunohistochemistry staining.The targeted binding relationship between miR-497 and signal transducer and activator of transcription 3 (STAT3) was detected by luciferase reporter assay.The relative expressions of miR-497, vascular endothelial growth factor A (VEGFA), tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1β and macrophage inflammatory protein (MCP)-1 mRNA were detected by real-time quantitative PCR.At 14 days following modeling, the expression of STAT3 and p-STAT3 proteins in mice corneal tissues was detected by Western blot.The use and care of animals complied with the ARVO statement.The study protocol was approved by the Ethics Committee of Renmin Hospital of Wuhan University (No.2019K-K010).Results:Corneal injury, inflammatory cell infiltration and CNV occurred in mice cornea after alkali burn.Corneal epithelial injury score, corneal stromal turbidity score and CNV area increased first and reached the peak on the 14th day after modeling, and then decreased.There were significant differences in corneal epithelial injury score, corneal stromal turbidity score, CNV area and number of CD31-positive cells among various time points after alkali burn ( Fgroup=49.19, 34.56, 44.56, 77.56; all at P<0.01; Ftime=51.62, 65.62, 71.32, 46.12; all at P<0.01). Corneal epithelial injury score, corneal stromal turbidity score, CNV area and the number of CD31-positive cells were greater in KO group at various time points than in WT and TG groups, and those in WT group were greater than in TG group (all at P<0.05). In WT STAT3 co-transfected cells, the luciferase activity of the miR-497 group was significantly lower than that of the miR-negative control group and normal control group (both at P<0.05). In mutant STAT3-transfected cells, there was no significant difference in luciferase activity among all groups ( F=0.69, P=0.56). On the 14th day after modeling, the relative expression levels of miR-497 in corneal tissue of WT, KO and TG groups were 0.68±0.11, 0.41±0.06 and 1.05±0.14, respectively, which were significantly lower than 1.00±0.04, 0.56±0.07 and 1.34±0.11 before modeling (all at P<0.01). The relative expressions of STAT3 and p-STAT3 were higher in KO group than in WT and TG groups, and were lower in TG group than in WT group, and the differences were statistically significant (all at P<0.05). The expressions of VEGFA, TNF-α, IL-6, IL-1β and MCP-1 mRNA at various time points after modeling in various groups were significantly higher than before modeling, which were higher in KO group than in WT and TG groups and were lower in TG group than in WT group, and the differences were statistically significant (all at P<0.01). Conclusions:MiR-497 inhibits corneal inflammation and CNV formation induced by alkali burn.It might inhibit the activation of the inflammation signal pathway via targeting STAT3.

Objective:To investigate the inhibitory effect of miR-497 on the corneal epithelial healing in diabetic mice and its possible mechanism.Methods:Forty healthy clean-grade wild-type C57BL/J6 mice were randomly divided into a blank control group and a model control group, with 20 mice in each group.Another 20 CRISPR/Cas9-mediated miR-497 knockout mice and miR-497 overexpression mice were taken as miR-497 knockout and miR-497 overexpression groups, respectively.The diabetes model was constructed by continuous intraperitoneal injection of streptozotocin (STZ) to the mice in model control, miR-497 knockout and miR-497 overexpression groups, and the mice in blank control group were injected with an equal amount of citrate buffer, followed by 8-week normal feeding.After the establishment of diabetes model, the corneal epithelial injury model was further constructed by scraping off part of the corneal epithelium with a central diameter of 2 mm.The corneal epithelial defect area of mice in 0, 12, 24 and 36 hours after corneal epithelial injury was observed by corneal fluorescein sodium staining.The expression of Wnt3a and β-catenin proteins in mice corneal tissues was detected by Western blot.The expression of miR-497 as well as the mRNA expression levels of cell proliferation-associated factor genes CyclinD1, c-Myc, and Ki-67 mRNA was detected by real-time quantitative fluorescence PCR.The targeting relationship between miR-497 and wnt3a was detected by a dual luciferase reporter gene assay.Human corneal epithelial cells (HCEC) were cultured in vitro and transfected with miR-497 mimics, miR-497 mimics negative control, miR-497 inhibitor, and miR-497 inhibitor negative control by Lipo8000 as miR-497 mimics group, mimics negative control group, miR-497 inhibitor group, andmiR-497 inhibitor negative control group, respectively, all of which were cultured in high glucose medium containing 25% glucose.Another two groups of HCEC were taken and cultured in medium containing 5% and 25% glucose as control and high glucose groups, respectively.The cell proliferation viability was determined by CCK8 method.The use and care of animals complied ith the ARVO statement.The study protocol was approved by the Ethics Committee of Renmin Hospital of Wuhan University (2019K-K010). Results:Eight weeks after STZ injection, the blood glucose of mice was significantly higher and the weight was significantly lower in each diabetic model group than those of blank control group (all at P<0.05). At 12, 24 and 36 hours after the corneal epithelial injury, the percentages of corneal epithelial defect area observed by slit-lamp microscopy in model control group were significantly higher than those in blank control group and miR-497 knockout group and lower than those in miR-497 overexpression group, and the differences were statistically significant (all at P<0.05). The relative expressions of wnt3a and β-catenin proteins in the corneal tissues of model control group were significantly lower than those of blank control group and miR-497 knockout group, but higher than those of miR-497 overexpression group, and the differences were statistically significant (all at P<0.05). The relative expressions of CyclinD1, c-Myc and Ki-67 mRNA in model control group were lower than those in miR-497 knockout group, but higher than those in miR-497 overexpression group, and the differences were statistically significant (all at P<0.05). The relative expression of miR-497 in model control group, miR-497 knockout group and miR-497 overexpression group was 1.00±0.02, 0.63±0.06 and 1.48±0.03, respectively, with a statistically significant difference ( F=19.62, P<0.01). The luciferase activity of miR-497-5p mimics group in wild-type wnt3a transfected cells was lower than that of miR-497-5p negative control group and empty vector group, and the differences were statistically significant (all at P<0.05). In the mutant wnt3a transfected cells, there was no significant difference in the luciferase activity among various groups ( F=0.73, P=0.59). The cell proliferation A value of high glucose group was 0.59±0.03, which was significantly lower than 0.59±0.03 of normal control group and 0.88±0.08 of miR-497 inhibitor group, but significantly higher than 0.48±0.11 of miR-497 mimics group (all at P<0.05). Conclusions:The silencing of miR-497 may promote the repair of diabetic corneal epithelial defects by targeting wnt/β-catenin pathway.