Objective:To investigate the effect and mechanism of Grifola frondosa extract on inflammatory response of colon tissue in rats with ulcerative colitis(UC)by regulating interleukin-6(IL-6)/Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway.Methods:Forty SD rats were randomly divided into blank control group,UC model group,Grifola frondosa treatment group,western medicine treatment group and combined treatment group,with 8 rats in each group.After UC rats were established by free drinking 3%DSS for 7 days,the treatment group were given Grifola frondosa extract 10 mg/(kg·d),sulfasalazine 0.3 g/(kg·d),and the same amount of two drugs,for 14 consecutive days.During the experiment,general state of rats were observed,and the disease activity index(DAI)score was calculated;pathological changes of rats colon tissue were observed by HE staining;protein expression levels of IL-6,JAK2,STAT3 and p-STAT3 in rats colon tissue were detected by Western blot;content of IL-6 in rats serum was detected by ELISA;protein contents and expressions of IL-6R and MPO in rats colon tissue were determined by immunohistochemistry.Results:Compared with blank control group,general state of rats in UC model group was poor,DAI score was increased,obvious tissue mucosal defects and inflammatory cell infiltration were observed by HE staining;protein expression levels of IL-6,JAK2,STAT3 and p-STAT3 in rats colon tissue and contents of IL-6R and MPO were significantly increased(P<0.01);content of IL-6 in rats serum was significantly increased(P<0.01),the difference was statistically significant.Compared with UC model group,general condition of rats in each treatment group was improved,DAI score was decreased,HE staining showed that mucosal defects were improved to varying degrees,and occasionally inflammatory cell infiltration was observed;protein expression levels of IL-6,JAK2,STAT3 and p-STAT3 in colon tissue were significantly decreased(P<0.01),contents of IL-6R and MPO in colon tissue and content of IL-6 in serum were significantly decreased(P<0.01 or P<0.05),the differences were statistically significant.Conclusion:Grifola frondosa extract can reduce the inflammatory response in colon tissue of UC rats by regulating expressions of IL-6/JAK2/STAT3 signaling pathway related factors.

Objective: To explore the clinical characteristics and prognosis of the new coronavirus 2019-nCoV patients combined with cardiovascular disease (CVD). Methods: A retrospective analysis was performed on 112 COVID-19 patients with CVD admitted to the western district of Union Hospital in Wuhan, from January 20, 2020 to February 15, 2020. They were divided into critical group (ICU, n=16) and general group (n=96) according to the severity of the disease and patients were followed up to the clinical endpoint. The observation indicators included total blood count, C-reactive protein (CRP), arterial blood gas analysis, myocardial injury markers, coagulation function, liver and kidney function, electrolyte, procalcitonin (PCT), B-type natriuretic peptide (BNP), blood lipid, pulmonary CT and pathogen detection. Results: Compared with the general group, the lymphocyte count (0.74×10⁹ (0.34×10⁹, 0.94×10⁹)/L vs. 0.99×10⁹ (0.71×10⁹, 1.29×10⁹)/L, P=0.03) was extremely lower in the critical group, CRP (106.98 (81.57, 135.76) mg/L vs. 34.34 (9.55,76.54) mg/L, P<0.001) and PCT (0.20 (0.15,0.48) μg/L vs. 0.11 (0.06,0.20)μg/L, P<0.001) were significantly higher in the critical group. The BMI of the critical group was significantly higher than that of the general group (25.5 (23.0, 27.5) kg/m² vs. 22.0 (20.0, 24.0) kg/m², P=0.003). Patients were further divided into non-survivor group (17, 15.18%) group and survivor group (95, 84.82%). Among the non-survivors, there were 88.24% (15/17) patients with BMI> 25 kg/m², which was significantly higher than that of survivors (18.95% (18/95), P<0.001). Compared with the survived patients, oxygenation index (130 (102, 415) vs. 434 (410, 444), P<0.001) was significantly lower and lactic acid (1.70 (1.30, 3.00) mmol/L vs. 1.20 (1.10, 1.60) mmol/L, P<0.001) was significantly higher in the non-survivors. There was no significant difference in the proportion of ACEI/ARB medication between the critical group and the general group or between non-survivors and survivors (all P>0.05). Conclusion COVID-19 patients combined with CVD are associated with a higher risk of mortality. Critical patients are characterized with lower lymphocyte counts. Higher BMI are more often seen in critical patients and non-survivor. ACEI/ARB use does not affect the morbidity and mortality of COVID-19 combined with CVD. Aggravating causes of death include fulminant inflammation, lactic acid accumulation and thrombotic events.

OBJECTIVE： To investigate the mechanism of calycosin （CA） inhibiting the proliferation and migration of lung adenocarcinoma cells by regulating miR-21/PTEN signaling pathway. METHODS： Using lung adenocarcinoma SPC-A1 cells as objects， cell proliferation was detected by MTT method after treated with different doses of CA （5， 15， 25， 50， 75， 100   μg/mL） for 12， 24， 48， 72 h. Cell survival rate， 30％ cell growth inhibition concentration （IC30） and half inhibition concentration （IC50） were calculated. Transwell migration test was used to detect the migration of cells after treated with low-dose， medium-dose and high-dose of CA （50， 75， 100 μg/mL） for 24 h. The number of stained cells was recorded and inhibition rate of cell migration were calculated. Western blotting assay and real-time PCR were used to detect the expression of miR-21 as well as the proteins and their mRNAs expression of PTEN， VEGF， MMP-9 after treated with low-dose， medium-dose and high-dose of CA （50， 75， 100 μg/mL） for 24 h. After transfected with miR-21 mimics and miR-21 inhibitor， the effects of CA （75 μg/mL） on the expression of miR-21 and the protein expression of PETN， VEGF and MMP-9 were detected. RESULTS： After treated with 50， 75， 100 μg/mL CA for 12， 24， 48 h， 25， 50， 75， 100 μg/mL CA for 72 h， cell survival rate was decreased significantly （P＜0.05 or P＜0.01）. IC30 of CA were 82.24， 50.45， 46.34， 31.81 μg/mL ； IC50 of CA were 108.06， 73.35， 70.08， 49.89 μg/mL during 12-72 h. Compared with normal control group， the number of stained cells in CA groups， protein expression of VEGF in CA low-dose group， expression of miR-21 as well as proteins and their mRNAs expression of VEGF， MMP-9 in CA medium-dose and high-dose groups were decreased significantly； the medium-dose and high-dose groups were significantly less or lower than low-dose group； the high-dose group was significantly less or lower than medium-dose group （P＜0.05 or P＜0.01）. Cell migration rate of CA groups as well as protein and its mRNA expression of PTEN in CA medium-dose and high-dose groups were increased significantly； the medium-dose and high-dose groups were significantly higher than the low-dose group； the high-dose group was significantly higher than the medium-dose groups （P＜0.05 or P＜0.01）. After transfected with miR-21 mimics， expression of miR-21 as well as protein expression of VEGF and MMP-9 were increased significantly in miR-21 mimic group， compared with normal control group； protein expression of PTEN was decreased significantly （P＜0.01）. After intervened by CA， expression of miR-21 as well as protein expression of VEGF and MMP-9 in cells were decreased significantly， compared with miR-21 mimic group； protein expression of PTEN was increased significantly （P＜0.05 or P＜0.01）. After transfected with miR-21 inhibitor， expression of miR-21 as well as  protein expression of VEGF and MMP-9 were decreased significantly in miR-21 inhibitor group， compared with normal control group； protein expression of PTEN was increased significantly （P＜0.05 or P＜0.01）. After intervened by CA， the expression of miR-21 and above protein had no significant change in cells， compared with miR-21 inhibitor group （P＞0.05）. CONCLUSIONS： CA can inhibit the proliferation and migration of lung adenocarcinoma SPC-A1 cells in a dose-dependent manner， which may be associated with the regulation of miR-21/PTEN signaling pathway.

Objective: To analyze pre- and post-operation electrocardiograms (ECGs) features of patients underwent orthotopic heart transplantation (OHT), and provide evidences for identifying and analyzing post OHT ECGs. Methods: Nine hundreds and ninty-eight pre- and post- OHT standard 12-leads ECGs from 110 consecutive patients, who underwent OHT in our hospital from May 2008 to May 2014, were analyzed. Results: The mean heart rate(HR)was (86.9±16.4) beats per minute before OHT, and (100.0±0.4) beats per minute after OHT. P wave′s amplitude, duration, amplitude multiplied by duration of donor heart in lead Ⅱ were (0.124±0.069)mV, (111.1±17.2)ms, (14.34±9.51)mV·ms before OHT; (0.054±0.037)mV, (86.9±27.0)ms, (5.02±4.03)mV·ms at 1 month after OHT; (0.073±0.049)mV, (93.9±17.5) ms, (7.00±4.81)mV·ms at 6 years after OHT. ECGs rotation occurred in 83.64%(92/110) patients after OHT, and prevalence of clockwise rotation was 76.36%(84/110). Sinus tachycardia was evidenced in 99.09%(109/110) patients after OHT, and incomplete right bundle branch block was present in 60.91%(67/110) patients after OHT. Pseudo complete atrioventricular block mostly occurred at 2 days after OHT. Prevalence of double sinus rhythm was 27.95%(263/941) post OHT, 40% of them occurred between the 1st and the 2nd month post OHT; the atrial rate of recipient hearts was (104.0±10.2) beats per minucte between the 3rd and the 6th month post OHT, and was (95.3±4.2) beats per minucte between the 4th year and the 5th year. P wave′s amplitude, duration, amplitude multiplied by duration of recipient heart in lead Ⅱ were (0.066±0.055) mV, (52.8±34.7) ms, (4.67±4.95) mV·ms at 1 month after OHT, (0.043±0.040)mV, (44.4±40.5) ms , (3.11±3.61) mV·ms between the 1st year and 2nd year after OHT. The absolute value of P-wave(originating from the donor heart) terminal force in chest leads increased in 48.99%(461/941) patients post OHT, the P-wave terminal force of V₁ , V₂ and V₃ were -0.044(-0.066, -0.028), -0.060(-0.087, -0.038), -0.035(-0.056, 0) mm·s. Notched P wave in chest leads was presented in 10.31%(97/941) patients post OHT. PR segment depression in chest leads occurred in 60.24%(100/166) patients between the 3rd month and the 6th month, the incidence of PR segment depression in V₁ , V₂ and V₃ was 21.04%(198/941), 37.41%(352/941) and 28.69%(270/941), respectively. Conclusions OHT is related to significantly changed ECGs. The mean HR increased significantly after OHT, then decreased gradually after half a year to one year, but it was still higher than preoperative mean HR after five or six years; the P waves of donor heart were usually inconspicuous or small in first month after OHT, and they became bigger after 2 months, and their duration and amplitude then became relatively steady afterwards. ECGs rotation, especially the clockwise rotation, was common post OHT. A variety of arrhythmias originating from the donor heart including sinus tachycardia and incomplete right bundle branch block could be found. Pseudo complete atrioventricular block could also be found in the early phase after OHT. With the extension of time, the incidence of double sinus rhythm reduced gradually. The atrial rate and P wave of recipient heart presented with a tendency to become lower. The absolute value of P-waves(originating from the donor heart) terminal force in chest leads (mainly V₁, V₂ and V₃) increased, notched P waves in chest leads (mainly V₁, V₂) and PR segments depression in chest leads (mainly V₂, V₃ and V₄) also belong to typical post OHT ECGs features.

Objective to observe the influence of Candida albicans infection on t cell subsets in mice with spleen deficiency,so as to explore the immune mechanism. Methods totally 150 healthy SPF mice were randomly divided into four groups:control group(N1 group,n = 30);Candida albicans infection group(N2,n = 40),spleen deficiency model group(M1,n = 40),spleen deficiency model combined with Candida albicans in-fection group(M2,n = 40). N2 and M1 mice were infected by Candida albicans at a concentration of 2×108 CFU/mL. ten mice were randomly se-lected from each group at 7,14,and 21 days after infection respectively,and the index was detected. Flow cytometry was used to detect the percent-age of CD4+/CD8+t cells in intestinal mucosa of mice,and the expression level of IL-4 and IFN-γ mRNA was detected by Rt-PCR assay. the levels of IFN-γ and IL-4 were detected by Western blot assay. Results Compared with N1 group,the proportion of CD4+t cells in the lamina of the natu-ral layer of the small intestine was decreased(P < 0.01),the proportion of CD8+t cells was increased(P < 0.01),the ratio of the two was significant-ly decreased(P < 0.01),and the expression levels of IL-4,mRNA IFN-γ and protein in the small intestine tissues were increased in other groups (P < 0.01). In addition,the expression level of IFN-γ was significantly increased in M2 group,while the expression of IL-4 was significantly in-creased in N2 group. Conclusion the susceptibility of Candida albicans infection was increased in spleen deficiency mice,which may be closely related to the regulation of th1/th2 balance.

Objective: To explore the relationship between expression of tumor necrosis factor-α( TNF-α) and electrophysiological heterogeneity in isolated heart tissues and isolated rat ventricular myocytes.The arrhythmogenic mechanisms of TNF-αwere further studied.Methods:Langendorff perfused heart tissues models were used to verify the arrhythmogenic effects of TNF-α.The monophasic action potentials( MAPs) of the endocardium and epicardium from the isolated heart tissues were recorded by elec-trophysiological experiments.The isolated rat ventricular myocytes were obtained by enzymatic dissociation.K+currents(Ito,IK1)were recorded by using whole cell patch clamp technique.Results: Compared to the control group, the difference in MAPD between endocardium and epicardium dramatically increased with TNF-α( P<0.05 ) .TNF-αcould cause MAP duration ( MAPD ) prolongation, and a single dose of TNF-αdifferentially affected the MAPs of endocardium and epicardium of isolated heart tissues.Compared to the control group,the K+currents(Ito,IK1)were dose-dependently decreased with TNF-αin rat ventricular myocytes(P<0.05).However, etanercept had no effects on the MAPD in the absence of TNF-α.Conclusion:TNF-α-induced heterogeneity of MAPD between the endo-cardium and epicardium may provide the substrate for the onset of ventricular arrhythmias during acute myocardial infarction.The effect might be associated with TNF-αcontribute to re-entrant ventricular arrhythmias which resulted from decreased K+currents(Ito,IK1).

Objective To explore the differences between normotrophic and hypertrophic scars of lip after the surgical repair of the unilateral complete cleft lip in density of microvessels and the pattern of angiogenesis.Methods Hypertrophic scars (n=11) and normotrophic scars (n=20) were collected after correction of deformity of the unilateral complete cleft lip,and the tissues were stained with haematoxylin and eosin,and immunostained with anti-CD34 antibody.The structure of scar was observed and the microvessels were counted according to the CD34 expression.Using ImageJ software,the capillary density and length of the major and minor axes were measured,and the major:minor axes ratio was calculated.Results By statistical analysis of the capillary density,the length of the major and minor axes and the major:minor axes ratio were measured;we clarified that there were more capillaries in hypertrophic scars (87.91 ± 5.95)/mm2 than in normotrophic scars (49.84 ± 7.05)/mm2,(P＜0.01),and the length of the major and minor axes of hypertrophic scars (38.36± 26.36)and (17.33±10.45) μm were longer than the normotrophic scars (13.77±9.56)and (9.00± 5.14) μm,(P＜0.05.) The major:minor axes ratio of hypertrophic scars (2.85±0.57) was higher than the normotrophic scars (2.85 ± 0.57) (P＜0.01).Conclusions The significant increase in the density of microvessels and the variation in the pattern and morphology of angiogenesis are related to the formation and development of scar after operation of upper lip.

Objective To observe the hypoglycemic mechanism of total saponins of Momordica charantia (MC) in type 2 diabetes mellitus (DM) rats. Methods Among the selected 60 male Specific Pathogen Free (SPF)rats, 8 were random- ly chosen as control group, while others were fed with high fat and high glucose diet following streptozotocin injection from caudal vein 8 weeks after to construct type 2 DM models. After the DM models were successfully built, rats were then ran- domized into five groups: DM control group (n=8), the metformin group (n=8) and three groups of total saponins of MC with different dosage (n=8 in each group). The total saponins of MC groups include DM rats administrated with total saponins of MC 100, 200, 400 mg/(kg·d) for 8 weeks, the metformin group include DM rats administrated with metformin 50 mg/(kg·d) for 8 weeks. At the end of the experiment, the fasting blood glucose and insulin were examed. At the same time, a part of pan- creas islet, liver and skeletal muscle were preserved. The pancreas islet structure, the hepatic glycogen and Glucose trans- porter 4 (GLUT4) expression were observed by electron microscope, glycogen dyeing and immunoblot respectively. Results After 8 weeks treatment, compared to type 2 DM control group, fasting blood glucose and insulin values in MC groups were reduced more obviously. However, skeletal muscle GLUT4 expression level, insulin granules and hepatic glycogen increased obviously in MC groups. Conclusion Total saponins of MC has hypoglycemic effect. It’s mechanisms maybe include pro- moting the hepatic glycogen synthesis, inhibiting the hepatic glycogen decomposition and promoting insulin sensitivity by in- creasing peripheral tissue’s GLUT4 expression.

Objective To observe the effects of eye acupuncture and body acupuncture on tumor necrosis factor-α(TNF-α) expression in the cerebral cortex of rats after ischemia-reperfusion injury,in order to investigate the differences in therapeutic function between eye acupuncture and body acupuncture against acute cerebral ischemia-reperfusion injury. Method 48 Rats established by suture method were randomly divided into control group,sham operation group,model group,eye-acupuncture in point area group,eye-acupuncture outside point area group and body acupuncture group. After reperfusion 24 h,the neurophysical behaviours were accessed by ZeaLonga neurophysical impairment marks;the levels of plasma TNF-αwere determined by ELISA method;the expression of ischemic cerebral cortex TNF-αmRNA was measured by RT-PCR method;the expression of ischemic cerebral cortex TNF-αprotein was detected by western blot. Results After reperfusion 24 h,compared with control group,neurologic impairment marks of eye-acupuncture therapy in point area group and body acupuncture group both decreased obviously (P<0.01),however there were no significant differences between the eye-acupuncture in point area group and body acupuncture group;the levels of plasma TNF-αin rat cerebral cortex after the eye acupuncture therapy and body acupuncture therapy were obviously decreased (P<0.01),however there were no significant differences between the eye-acupuncture in point area group and body acupuncture group; the expressions of TNF-α mRNA and protein in rat cerebral cortex after the eye acupuncture therapy and body acupuncture therapy were also obviously down-regulated (P<0.01),however there were no significant differences between the eye-acupuncture in point area group and body acupuncture group.Conclusion The eye and body acupuncture therapy show the same effects on treating cerebral ischemia reperfusion. The mechanisms of these two therapies may be related to up regulating TNF-αexpression in rat cerebral cortex with ischemia-reperfusion injury.

BACKGROUND:Significant increasing of intercel ular adhesion molecule 1 expression can promote the occurrence of inflammatory response and increase brain tissue injury.
OBJECTIVE:To investigate the effect of eye acupuncture point and non-point therapy on intercel ular adhesion molecule 1 expression in rat hippocampus of acute cerebral ischemia-reperfusion injury model.
METHODS:Forty Sprague Dawley rats were divided into normal group, sham-operation group, model group, eye acupuncture point group and eye acupuncture non-point group. Rats in the model group, eye acupuncture point group and eye acupuncture non-point group were used to establish the acute cerebral ischemia-reperfusion injury model with suture method, the suture was deep for 1.8-2.2 cm. In the eye acupuncture point group, the acupuncture points of hepatic region, upper energizer area, lower energizer area and kidney area were selected when cerebral ischemia-reperfusion occurred immediately and 30 minutes before drawing materials, then horizontal needling the acupuncture points around the orbit for 20 minutes. In the eye acupuncture non-point group, the puncture site was selected at 3 mm away from eye acupuncture non-point area, and then needled the puncture site with the same method in the eye acupuncture point group.
RESULTS AND CONCLUSION:After eye acupuncture point therapy, the neurological deficit scores were decreased, and the intercel ular adhesion molecule 1 expression in rat hippocampus was significantly decreased (P<0.01). After eye acupuncture non-point therapy, there were no significant changes in neurological deficit scores and the expression of intercel ular adhesion molecule 1 protein and mRNA in rat hippocampus. The results indicate that eye acupuncture can significantly improve the rat cerebral ischemia-reperfusion injury, and the mechanism may relate with reducing the intercel ular adhesion molecule 1 expression in rat hippocampus.