Objective: To investigate the effects of salvianolic acid B (Sal B) on the quality of stored red blood cells (RBC) and to preliminarily explore its underlying mechanisms. Methods: Whole blood was collected from healthy adult volunteers, and suspended RBC were prepared and stored in MAP (mannitol-adenine-phosphate) solution. The RBC suspensions were divided into a control group and a Sal B group, with the latter supplemented with Sal B at a final concentration of 10 μM. All samples were mixed and stored at 2-6℃. RBC were collected on days 0, 7, 14, 21, 28, and 35 for evaluation. RBC damage was assessed through cell counting, morphological observation, osmotic fragility test, and hemolysis rate measurement. Cytosolic ROS levels and intracellular SOD activity were measured to evaluate oxidative stress. Results: Sal B helped maintain RBC morphology and osmotic stability, and reduced the generation of morphologically abnormal cells such as spherocytes and echinocytes. On days 28 and 35, the hemolysis rate in the Sal B group was significantly lower than that in the control group. Compared with the control group, the Sal B group exhibited decreased cytosolic ROS levels and increased intracellular SOD activity from day 14 onward. Conclusion: Sal B alleviates RBC storage lesion by reducing oxidative stress.

Objective To investigate the effects of lumbar epidural block of renal sympathetic nerve on cardiac function and serum inflammatory factors in rats with myocardial infarction.Methods Thirty healthy male Sprague Dawley rats were divided into asham operation group,myocardial infarction group and sympathetic nerve block group according to the random number table method,with 10 rats in each group.Lumbar epidural catheterization was applied in all rats in the 3 groups.After catheter insertion,myocardial infarction was induced by ligation of the anterior descending branch of the left coronary artery of rats in the myocardial infarction group and the sympathetic nerve block group,while the left anterior descending branch of rats in the sham operation group was not ligated.After myocardial infarction,the rats in the sympathetic nerve block group were injected with 50 μL of ropivacaine(volume fraction:0.2％)via a lumbar epidural catheteronce every 24 hours,and the injection lasted until 14 days after surgery.In the myocardial infarction group,50 μL of 9 g·L-1sodium chloride was injected into the rats through a lumbar epidural catheteronce every 24 hours,and the injection lasted until 14 days after surgery.Rats in the sham group did not receive the injection of any drugs.At 14 days after operation,echocardiography was performed on rats in the 3 groups to measure ejection fraction(EF),left ventricular short-axis shortening rate(FS)and cardiac output(CO).A total of 1 mL of cervical venous blood was extracted from each rat in the 3 groups at 24 h after surgery,and 5 mL of abdominal aortic blood was extracted from rats in the 3 groups at 14 days after surgery.Serum levels of norepinephrine(NE),cardiac troponin I(cTnl),interleukin-18(IL-18),interleukin-1β(IL-1 β)and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay.At 14 days after surgery,the rats in the 3 groups were anesthetized by intraperitoneal injection of 100 g·L-1 chloral hydrate(300 mg·kg-1),and the thoracic cavity of the rats was opened to remove the heart.The myocardial infarction area of the rats was detected by 2,3,5-triphenyltetrazolium chloride staining,and the proportion of the myocardial infarction area was calculated.Results On the 14th day after surgery,EF,FS and CO of rats in the myocardial infarction group and sympathetic nerve block group were significantly lower than those in the sham operation group,while EF,FS and CO of rats in the sympathetic nerve block group were significantly higher than those in the myocardial infarction group(P＜0.05).The serum cTnI level of rats in the myocardial infarction group and sympathetic nerve block group was significantly higher than that in the sham operation group(t=68.260,15.110;P＜0.05),and the serum cTnI level of rats in the myocardial infarction group was significantly higher than that in the sympathetic nerve block group(t=27.920,P＜0.05).The proportion of the myocardial infarction area of rats in the sympathetic nerve block group was significantly lower than that in the myocardial infarction group(t=14.182,P＜0.001).There was no significant difference in the serum NE,IL-18,IL-1 β and TNF-α levels of rats between 24 hours and 14 days after surgery in the sham operation group(P＞0.05).The serum NE,IL-1 β and TNF-αlevels of rats at 14 days after surgery were significantly higher than those at 24 hours after surgery,and the IL-18 level of rats was significantly lower than that at 24 hours after surgery in the myocardial infarction group(P＜0.05).The serum NE,IL-18 and TNF-α levels of rats at 14 days after surgery were significantly lower than those at 24 hours after surgery,and the IL-1 βlevel of rats was significantly higher than that at 24 hours after surgery in the sympathetic nerve block group(P＜0.05).At 24 hours and 14 days after surgery,the serum NE,IL-18,1L-1 β and TNF-α levels of rats in the myocardial infarction group and sympathetic nerve block group were significantly higher than those in the sham operation group,while the serum NE,IL-18,IL-1 β and TNF-α levels of rats in the sympathetic nerve block group were significantly lower than those in the myocardial infarction group(P＜0.05).Conclusion Lumbar epidural block of renal sympathetic nerve in rats can significantly reduce the levels of serum inflammatory factors and NE,and improve cardiac function.

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.

Objective:To evaluate the effect of superior cervical ganglion block (SCGB) on cardiac function and nucleotide like receptor protein 3 (NLRP3) signaling pathway in a rat model of myocardial ischemia-reperfusion (I/R).Methods:Sixty healthy SPF male Sprague-Dawley rats, weighing 250-300 g, aged 2-3 months, were divided into 4 groups ( n=15 each) using a random number table method: sham operation group (sham group), myocardial I/R group (IR group), myocardial I/R + normal saline group (IR+ NS group), and myocardial I/R + SCGB group (IR+ SCGB group). Myocardial I/R model was developed by ligation of the left anterior descending branch of the coronary artery for 45 min followed by restoration of blood flow in anesthetized aninals. IR+ SCGB group received SCGB (0.25% ropivacaine 0.1 ml) at 10 min before reperfusion once a day for 2 consecutive weeks, while 0.9% sodium chloride was given instead of ropivacaine in IR+ NS group. Blood samples were collected at 24 h and 14 days of reperfusion for determination of serum concentrations of norepinephrine (NE), troponin T (TnT), tumor necrosis factor-alpha (TNF-α), interleukin-18 (IL-18) and IL-1β by enzyme-linked immunosorbent assay. Echocardiography was performed before ischemia and at 14 days of reperfusion, and left ventricular short axis shortening rate (FS), ejection fraction (EF), and cardiac output (CO) were measured. The rats were sacrificed at 14 days of reperfusion and the hearts were taken for determination of the contents of norepinephrine (NE) in myocardial tissues in the infarction area (by enzyme-linked immunosorbent assay), percentage of myocardial fibrosis area (by Masson staining), M1 macrophage marker CD68 + cell count in the infarction area (by immunohistochemical method), and expression of NLRP3 and gasdermin D (GSDMD) in myocardial tissues (by Western blot). Results:Compared with Sham group, the serum concentrations of TnT, TNF-α, IL-18 and IL-1β, percentage of myocardial fibrosis area, and NE levels in serum and myocardial tissues were significantly increased, the expression of NLRP3 and GSDMD in myocardial tissues was up-regulated, CD68 + cell count was increased, and EF, CO and FS were decreased in IR group ( P<0.05). Compared with IR group, the serum concentrations of TnT, TNF-α, IL-18 and IL-1β, percentage of myocardial fibrosis area, and NE levels in serum and myocardial tissues were significantly decreased, the expression of NLRP3 and GSDMD in myocardial tissues was down-regulated, CD68 + cell count was decreased, and EF, CO and FS were increased in IR+ SCGB group ( P<0.05), and no statistically significant changes were found in the parameters mentioned above in IR+ NS group ( P>0.05). Conclusions:SCGB can improve the cardiac function in a rat model of myocardial I/R, and the mechanism may be related to the inhibition of NLRP3 signaling pathway.

Objective: Investigate osteogenic differentiation of canine periodontal ligament stem cells ( cPDLSCs) via over-expression ephrinB2 in cPDLSCs． Methods : cPDLSCs were isolated from the premolars and molars of Beagle．After transfected with EfnB2-GFP-Bsd and GFP-Bsd empty Vector，cPDLSCs were induced to osteogenic differentiation．Western blot was used to invest the expression of ephrinB2 protein．The effect of osteogenic differentiation of EfnB2-cPDLSCs and Vector-cPDLSCs were analyzed by RT-PCR , CCK-8，Alizarin-red S staining and ALP. Results: There was no significant difference in cell proliferation between EfnB2-cPDLSCs and Vector-cPDLSCs．While EfnB2-cPDLSCs displayed an enhanced ALP activity and more prominent mineralized nodules compared with Vector-cPDLSCs．The odonto-/ osteogenic genes in EfnB2-cPDLSCs were also highly enhanced． Conclusion The results of our study indicated that ephrinB2 gene-transfected cPDLSCs showed enhanced osteogenic differentiation.

OBJECTIVETo understood the accuracy of oral fluid-based rapid HIV self-testing among men who have sex with men (MSM) and related factors.

METHODSSurvey was conducted among MSM selected through non-probability sampling to evaluate the quality of their rapid HIV self-testing, and related information was analyzed.

RESULTSThe most MSM were aged 21-30 years (57.0%). Among them, 45.7% had educational level of college or above, 78.5% were unmarried, 59.3% were casual laborers. The overall accuracy rate of oral fluid based self-testing was 95.0%, the handling of"inserting test paper into tube as indicated by arrow on it"had the highest accuracy rate (98.0%), and the handling of"gently upsetting tube for 3 times"had lowest accuracy rate (65.0%); Chi-square analysis showed that educational level, no touch with middle part of test paper, whether reading the instruction carefully, whether understanding the instruction and inserting test paper into tube as indicated by the arrow on it were associated with the accuracy of oral fluid-based rapid HIV self-testing, (P<0.05). Multivariate logistic regression analysis indicated that educational level, no touch with middle part of test paper and understanding instructions were associated with the accuracy of oral fluid-based rapid HIV self-testing.

CONCLUSIONSThe accuracy of oral fluid-based rapid HIV self-testing was high among MSM, the accuracy varied with the educational level of the MSM. Touch with the middle part of test paper or not and understanding the instructions or not might influence the accuracy of the self-testing.

Adult ; Demography ; HIV Infections ; diagnosis ; Homosexuality, Male ; Humans ; Male ; Mass Screening ; methods ; Saliva ; Self Care ; Surveys and Questionnaires ; Young Adult

Objective To investigate the effect of diuretics combined with the new class volume-removing drugs nesiritide and tolvaptan on refractory heart failure (RHF).Methods We collected 78 patients with RHF from cardiovascular ICU in our hos-pital,and patients were divided into combined volume-removing drugs treatment group (CVR group)and traditional diuretics com-bined with inotropic treatment group (control group).We observed the differences in the heart failure symptoms control rate and the average hospitalization days between the two groups.The difference in average urine volume and weight loss during hospitaliza-tion was observed.Results The heart failure symptoms were controlled and successful discharge in 89.7% of patients in CVR group,which was better than that in control group(71.8%).The corresponding rates of death or abandonment was decreased sig-nificantly in CVR group,and the average number of hospitalization days in CVR group was significantly shorter than that in control group(P <0.05).Meanwhile,the average daily urine volume and the weight loss decreased significantly in CVR group than those in control group(P <0.05).Conclusion The therapy strategy of combined volume-removing drugs can significantly improve the vol-ume removing and control rate of heart failure symptoms.

Objective To describe the anatomical variants of infants’internal cerebral veins and their tributaries with MR suscep-tibility weighted imaging,and to evaluate the capability in the visualization of cerebral deep veins.Methods 80 healthy infants were enrolled in this study.All the brain images were obtained by a 3D gradient-echo sequence (Enhanced T2 ? weighted angiography-ES-WAN)on a 3.0T MR,which were reconstructed with minimal intensity projection.The septal vein,thalamostriate vein,internal cerebral vein and anterior caudate nucleus veins were evaluated.4 types (ⅠA,ⅠB,ⅡA,ⅡB)were classified based on their rela-tionship with the septal vein-internal cerebral vein junction and interventricular foramen,and 3 types (Ⅰ,Ⅱ,Ⅲ)based on the drainage patterns of the anterior caudate nucleus veins.Results The septal vein,thalamostriate vein and internal cerebral vein could be clearly and continuously visualized in 100% infants,and the visualization rate of the anterior caudate nucleus veins was 88.1%. TypeⅠA and TypeⅠ were most common in the two classification patterns.Conclusion Infants’internal cerebral veins and their tributaries are able to be clearly shown with the minimal intensity projection in susceptibility weighted imaging,which is a good method to evaluate the cerebral deep veins in infants.

Objective To survey the mental health status of rural-to-urban migrant workers in Shanghai.Methods The survey subjects were selected from migrant workers in Shanghai with a multi-stage stratified cluster sampling method.The Symptom Check List ( SCL-90 ) scores were used for evaluation of metal health status.Results The prevalence of mental health problem of 5 626 rural-to-urban migrant workers in Shanghai was 18.8% ( 1 058/5 626 ).The mean total score of SCL-90 was 114.86 ±31.21.Compulsive-obsessive, interpersonal sensitivity, hostility ranked the top three among 9 factors, with scores of 1.37 ±0.43, 1.31 ±0.43 and 1.30 ±0.41 respectively.The mean total scores and scores of 9 factors of rural-to-urban migrant workers in Shanghai were lower than those of Chinese adults norm the mean total scores(129.96 ±38.76) ( t =-15.34, -3.84 --24.08 respectively, all P <0.01) .There were significant differences in mental health among migrant workers with different age , education , marital status , living conditions and occupation (all P<0.05).Conclusion The prevalence of mental health problems in Shanghai rural-to-urban migrant workers is high, with are related to their age, education, marital status, living conditions and occupation.