Intervention effect of Chuanxiong-Chishao herb pair on circRNA/lncRNA expression profile in a myocardial infarction-atherosclerosis model / 中国中药杂志

This study aimed to explore the intervention effect of Chuanxiong-Chishao herb pair(CX-CS) on a myocardial infarction-atherosclerosis(MI-AS) mouse model and investigate its effect on the expression profile of circular RNAs(circRNAs)/long non-coding RNAs(lncRNAs) in ischemic myocardium and aorta. Sixty male ApoE~(-/-) mice were randomly assigned to a model group, high-, medium-, and low-dose CX-CS groups(7.8, 3.9, and 1.95 g·kg~(-1)), and a positive drug group(metoprolol 26 mg·kg~(-1) and simvastatin 5.2 mg·kg~(-1)), with 12 mice in each group. Male C57BL/6J mice were assigned to the sham group. The mice in the model group and the groups with drug intervention were fed on a high-fat diet for 10 weeks, followed by anterior descending coronary artery ligation. After that, the mice were fed on a high-fat diet for another two weeks to induce the MI-AS model. The mice in the sham group received normal feed, followed by sham surgery without coronary artery ligation. Mice in the groups with drug intervention received CX-CS or positive drug by gavage for four weeks from the 9th week of high-fat feeding, and those in the model group and the sham group received an equal volume of normal saline. Whole transcriptome sequencing was performed on the heart and aorta tissues of the medium-dose CX-CS group, the model group, and the sham group after administration. The results showed that the medium-and high-dose CX-CS groups showed improved cardiac function and reduced myocardial fibrosis area, and the medium-dose CX-CS group showed significantly reduced plaque area. CX-CS treatment could reverse the expression of circRNA＿07227 and circRNA＿11464 in the aorta of AS model and circRNA expression(such as circRNA＿11505) in the heart of the MI model. Differentially expressed circRNAs between the CX-CS-treated mice and the model mice were mainly enriched in lipid synthesis, lipid metabolism, lipid transport, inflammation, and angiogenesis in the aorta, and in angiogenesis, blood pressure regulation, and other processes in the heart. CX-CS treatment could reverse the expression of lncRNAs such as ENSMUST00000162209 in the aorta of the AS model and TCONS＿00002123 in the heart of the MI model. Differentially expressed lncRNAs between the CX-CS-treated mice and model mice were mainly enriched in lipid metabolism, angiogenesis, autophagy, apoptosis, and iron death in the aorta, and in angiogenesis, autophagy, and iron death in the heart. In summary, CX-CS can regulate the expression of a variety of circRNAs and lncRNAs, and its intervention mechanism in coronary heart disease may be related to the regulation of angiogenesis and inflammation in ischemic myocardium, as well as lipid metabolism, lipid transport, inflammation, angiogenesis in AS aorta.

Effective fraction from Simiao Wan prevents hepatic insulin resistant by inhibition of lipolysis via AMPK activation / 中国天然药物

Simiao Wan (SMW) is a traditional Chinese formula, including Atractylodis Rhizoma, Achyranthis Bidentatae Radix, Phellodendri Chinensis Cortex and Coicis Semen at the ratio of 1:1:2:2. It can be used to the treatment of diabetes. However, its bioactive compounds and underlying mechanism are unclear. This study aimed to screen the antilipolytic fraction from SMW and investigate its therapeutic mechanisms on hepatic insulin resistance. Different fractions of SMW were prepared by membrane separation combined with macroporous resin and their antilipolytic activities were screened in fasted mice. The effects of 60% ethanol elution (ESMW) on lipolysis were investigated in 3T3-L1 adipocytes stimulated by palmitic acid (PA) and high fat diet (HFD)-fed mice. In our study, ESMW is the bioactive fraction responsible for the antilipolytic activity of SMW and 13 compounds were characterized from ESMW by UHPLC-QTOF-MS/MS. ESMW suppressed protein kinase A (PKA)-hormone-sensitive lipase (HSL) related lipolysis and increased AMP-activated protein kinase (AMPK) phosphorylation in PA challenged 3T3-L1 adipocytes. AMPKα knockdown abolished the inhibitory effects of ESMW on IL-6 and HSL pSer-660, revealing that the antilipolytic and anti-inflammatory activities of ESMW are AMPK dependent. Furthermore, ESMW ameliorated insulin resistance and suppressed lipolysis in HFD-fed mice. It inhibited diacylglycerol accumulation in the liver and inhibited hepatic gluconeogenesis. Conditional medium collected from ESMW-treated 3T3-L1 cells ameliorated insulin action on hepatic gluconeogenesis in liver cells, demonstrating the antilipolytic activity contributed to ESMW beneficial effects on hepatic glucose production. In conclusion, ESMW, as the antilipolytic fraction of SMW, inhibited PKA-HSL related lipolysis by activating AMPK, thus inhibiting diacylglycerol (DAG) accumulation in the liver and thereby improving insulin resistance and hepatic gluconeogenesis.

The role and application of organ reserve capacity in sepsis / 中华消化外科杂志

Sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection. The injured organ is the main target of treatment. Organic functional reserve refers to the ability of an organ or system to return to its original physiological state following acute physiological stress or pathological injury, which has not received widespread attention. The organ reserve capacity is expected to complement the existing sepsis-related scoring system to optimize disease severity grading and evaluate prognosis. Source control, appropriate using of antibiotics and organ supporting can reduce further damage of organ reserve capacity, while nutritional therapy and rehabilitation may enhance it. Therefore, the authors believe that in further basic theoretical research and clinical practice, more attention can be paid to the monitoring and management of organ reserve capacity in sepsis, which may help improving the diagnosis and treatment of sepsis and prognosis of patients.

Preparation of purified proteins from fresh Pheretima and their inhibitory effect against pulmonary fibrosis in mice / 南方医科大学学报

OBJECTIVE@#To develop a convenient method for rapid purification of fresh Pheretima proteins and assess the inhibitory effect of these proteins against pulmonary fibrosis.@*METHODS@#The crude extract of fresh Pheretima was obtained by freeze-drying method and then purified by size exclusion chromatography. The composition of the purified proteins was analyzed by mass spectrometry. MRC-5 cells were treated with 5 ng/mL TGF-β1 alone (model group) or in combination with SB431542 (2 μmol/L) or the purified proteins (13.125 μg/mL), and the cytotoxicity of purified proteins and their inhibitory effects on cell proliferation were detected with CCK8 assay. Flow cytometry was used to detect the changes in cell apoptosis, and the cellular expressions of α-SMA, Vimentin, E-cadherin, collagen I, Smad2/3 and P-Smad2/3 were detected using RT-PCR and Western blotting. In the animal experiment, adult male C57BL/6 mice were subjected to intratracheal instillation of bleomycin followed by treatment with the purified proteins (5 mg/mL) for 21 days, after which HE and Masson staining was used to observe the pathological changes in the lung tissue of the mice.@*RESULTS@#We successfully obtained purified proteins from fresh Pheretima protein by size exclusion chromatography. Treatment with the purified proteins significantly inhibited TGF-β1-induced proliferation of MRC-5 cells (P < 0.01), reduced the cellular expressions of α-SMA, Vimentin and collagen I (P < 0.001 or P < 0.01), increased the expression of E-cadherin (P < 0.01), and inhibited the expressions of Smad2/3 and P-Smad2/3 (P < 0.001 or P < 0.01). In male C57BL/6 mice models of bleomycin-induced pulmonary fibrosis, treatment with the purified proteins obviously reduced the number of inflammatory cells and fibrotic area in the lungs.@*CONCLUSION@#The purified proteins from fresh Pheretima obtained by size exclusion chromatography can inhibit pulmonary fibrosis in mice by regulating the TGF-β/ Smad pathway.

Effects of components in stasis-resolving and collateral-dredging Chinese herbal medicines on angiogenesis and inflammatory response of human umbilical vein endothelial cells induced by VEGF / 中国中药杂志

The present study investigated the mechanism of components in stasis-resolving and collateral-dredging Chinese herbal medicines, including scutellarin(Scu), paeonol(Pae), and hydroxy safflower yellow A(HSYA), in the treatment of psoriasis by regulating angiogenesis and inflammation. The human umbilical vein endothelial cells(HUVECs) cultured in vitro were divided into a normal group, a model group, a VEGFR tyrosine kinase inhibitor Ⅱ(VRI) group, and Scu, Pae, and HSYA groups with low, me-dium, and high doses. Cell viability was detected by the CCK-8 assay. Cell migration was detected by wound healing assay. Tube formation assay was used to measure the tube formation ability. Western blot was used to detect the protein expression of the VEGFR2/Akt/ERK1/2 signaling pathway. The secretion levels of inflammatory cytokines IFN-γ, IL-1β, IL-6, and TNF-α were detected by ELISA. The results showed that compared with the model group, all the Scu, Pae, and HSYA groups could reduce cell viability, inhibit cell migration and tube formation(P<0.05, P<0.01), and down-regulated the protein expression of VEGFR2, p-VEGFR2, Akt, p-Akt, ERK1/2, and p-ERK1/2. Scu and Pae could down-regulate VEGFR2 expression(P<0.05, P<0.01), while other groups only showed a downward trend. Scu and Pae significantly reduced IFN-γ and IL-6 levels(P<0.01), and HSYA significantly reduced the levels of IFN-γ, IL-1β, and IL-6(P<0.01). Scu, Pae, and HSYA had no significant effect on TNF-α. The results suggested that Scu, Pae, and HSYA may exert a therapeutic role in psoriasis-related angiogenesis and inflammation by inhibiting VEGFR2/Akt/ERK1/2 signaling pathway and inhibiting the secretion of IFN-γ, IL-1β, and IL-6.

Role of crenel lateral lumbar interbody fusion in the precise selection of fusion level in posterior correction for adult degenerative scoliosis / 中华外科杂志

Objective: To investigate the role of minimally invasive crenel lateral lumbar interbody fusion (CLIF) in the decision of fusion level in posterior correction for severe adult degenerative scoliosis. Methods: This is a prospective study.Patients with level Ⅴ and Ⅵ of Lenke-Silva classification who were treated at Department of Orthopedics,the Second Affiliated Hospital, School of Medicine, Zhejiang University from June 2016 to March 2019 were included.First,the enrolled patients completed the preoperative clinical and imaging examination,the Lenke-Silva classification was evaluated,the surgical segments in first-stage CLIF was determined and the fusion segments required for single-stage posterior correction was predicted.After the first-stage CLIF,patients received reassessment of Lenke-Silva classification and global coronal and sagittal balance.Patients were divided into two groups:the effective group (level of Lenke-Silva classification decreased) and the ineffective group (level of Lenke-Silva classification unchanged).Second-stage posterior surgery was performed based on the results of reassessments.The fusion segment,Cobb angle,parameters of global coronal and sagittal plane,visual analogue pain score (VAS) and Oswestry disability index (ODI) were compared between the two groups preoperatively,after first-stage CLIF,second-stage posterior fixation and at the final follow-up.The potential factors associated with the decrease of the level of Lenke-Silva classification were recorded and compared between the two groups.Independent sample t test,repeated measure analysis of variance,rank sum test,χ2 test or Fisher exact method were used to compare the difference among groups. Results: Fifty-four patients were enrolled,including 8 males and 46 females,aged (68.8±5.8) years (range:56 to 77 years).Preoperatively,26 patients were classified as level Ⅴ by Lenke-Silva classification,28 cases were grade Ⅵ.CLIF was performed in 194 segments,with 114 segments(58.8%) receiving anterior column realignment (ACR) and 15 segments(7.7%) using hyperlordotic cages.After first-stage CLIF,22 patients with level Ⅴ and 10 patients with Ⅵ of Lenke-Silva classification decreased and were classified into effective group.The level of the remaining 4 patients with level Ⅴ and 18 patients with grade Ⅵ unchanged and were classified into ineffective group.Preoperatively,the apical vertebrae was below L1 in all 32 patients of effective group and 18 (81.8%,18/22) patients of ineffective group.The difference was statistically significant (P=0.023).There were 7 (31.8%,7/22) patients had continuous osteophyte in front of the intervertebral space in ineffective group,while none patient had continuous osteophyte in front of the intervertebral space in effective group,and the difference was statistically significant (P=0.001).In first-stage CLIF,more intraoperative ACR(71.2% vs.39.5%,χ²=20.660,P<0.01)and hyperlordotic cage (12.7% vs.0,P=0.001) were used in the effective group,while there was less severe cage subsidence after the operation (5.9% vs.15.8%,χ²=4.793,P=0.029) in effective group.After first-stage CLIF,there was no difference in the Cobb angle between the two groups.While,lumbar lordosis (LL) in effective group (34.0±8.3)° was greater than that of the ineffective group (25.5±9.7)° (t=3.478,P=0.001),and the difference between the pelvic incidence (PI) and LL in effective group (15.7±4.6)°was significantly smaller than ineffective group(20.0±10.8)° (t=-2.129,P=0.038).The posterior fusion levels was less,the rate of fusion to thoracic spine region and the actual fusion segment was less than that of single-stage posterior correction in effective group (all P<0.01).All patients were follow-up for 24 to 45 months.There was no significant difference in radiological and clinical results between the two groups after first-,second-stage surgery and at the final follow-up (all P>0.05). Conclusions: First-stage CLIF decreased the Lenke-Silva classification of some patients with severe degenerative scoliosis.Combined with the reassessment of Lenke-Silva classification and global coronal and sagittal plane,it helps to accurately determine the fusion segment.Decrease of Lenke-Silva classification is associated with the preoperative level of apical vertebrae,continuous osteophytes in front of the intervertebral space,intraoperative use of ACR and hyperlordotic cage and the degree of cage subsidence postoperatively.

Correlation of Platelet and Coagulation Function with Blood Stasis Syndrome in Coronary Heart Disease: A Systematic Review and Meta-Analysis / 中国结合医学杂志

OBJECTIVE@#To investigate the correlation of platelet and coagulation function with blood stasis syndrome (BSS) in coronary heart disease (CHD).@*METHODS@#The protocol for this meta-analysis was registered on PROSPERO (CRD42019129452). PubMed, Excerpta Medica Database (Embase), the Cochrane Library, and China National Knowledge Infrastructure (CNKI) were searched from inception to 1st June, 2020. Trials were considered eligible if they enrolled BSS and non-BSS (NBSS) patients with CHD and provided information on platelet and coagulation function. The platelet function, coagulation function, and fibrinolytic activity were compared between the BSS and NBSS groups. Forest plots were generated to show the SMDs or ESs with corresponding 95% CIs for each study. Subgroup analysis and sensitivity analysis were performed to explore potential sources of heterogeneity.@*RESULTS@#The systematic search identified 1,583 articles. Thirty trials involving 10,323 patients were included in the meta-analysis. The results showed that mean platelet volume, platelet distribution width, platelet aggregation rate, platelet P selectin, fibrinogen, plasminogen activator inhibitor-1 (PAI-1), thromboxane B2 (TXB2), 6-keto-prostaglandin F1alpha (6-keto-PGF1 α), and TXB2/6-keto-PGF1 α were higher in the BSS group than in the NBSS group (P<0.05 or P<0.01). Activated partial thromboplastin time was lower in the BSS group than in the NBSS group in the acute phase of CHD (P<0.01). The R and K values in thromboelastography and tissue plasminogen activator (t-PA) and t-PA/PAI-1 were lower in the BSS group than in the NBSS group (all P<0.01). No difference was found in the results of platelet count, plateletcrit, maximum amplitude, von Willebrand factor, prothrombin time, thrombin time, international normalized ratio, etc. between groups.@*CONCLUSIONS@#Increased platelet function, hypercoagulability, and decreased fibrinolytic activity were found among CHD patients with BSS.

Danqi Tablet () Regulates Energy Metabolism in Ischemic Heart Rat Model through AMPK/SIRT1-PGC-1α Pathway / 中国结合医学杂志

OBJECTIVE@#To investigate the cardioprotective effect of Danqi Tablet (DQT, ) on ischemic heart model rats and the regulative effect on energy metabolism through peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α).@*METHODS@#Rat ischemic heart model was induced by ligation of left anterior descending coronary artery. Totally 40 Sprague-Dawley rats were randomly divided into sham group, model group, DQT group (1.5 mg/kg daily) and trimetazidine (TMZ) group (6.3 mg/kg daily) according to a random number table, 10 rats in each group. Twenty-eight days after continuous administration, cardiac function was assessed by echocardiography and the structures of myocardial cells were observed by hematoxylin-eosin staining. The level of adenosine triphosphate (ATP) in myocardial cells was measured by ATP assay kit. Expressions level of key transcriptional regulators, including PGC-1α, Sirtuin 1 (SIRT1), AMP-activated protein kinase (AMPK), and downstream targets of PGC-1α, such as mitofusin 1 (MFN1), mitofusin 2 (MFN2) and superoxide dismutase 2 (SOD2) were measured by Western blot. Expression level of PGC-1α was examined by immunohistochemical staining.@*RESULTS@#The rat ischemic heart model was successfully induced and the heart function in model group was compromised. Compared with the model group, DQT exerted cardioprotective effects, up-regulated the ATP production in myocardial cells and inhibited the infiltration of inflammatory cells in the margin area of infarction of the myocardial tissues (P<0.01). The expressions of PGC-1α, SIRT1 and AMPK were increased in the DQT group (all P<0.05). Furthermore, the downstream targets, including MFN1, MFN2 and SOD2 were up-regulated (P<0.05 or P<0.01). Compared with the TMZ group, the expression levels of PGC-1α, MFN1 and SOD2 were increased by DQT treatment (P<0.05 or P<0.01).@*CONCLUSION@#DQT regulated energy metabolism in rats with ischemic heart model through AMPK/SIRT1 -PGC-1α pathway. PGC-1α might serve as a promising target in the treatment of ischemic heart disease.

Value of clinical staging and typing of hilar cholangiocarcinoma in evaluating radical resection and prognosis / 临床肝胆病杂志

Hilar cholangiocarcinoma (HCCA) is a malignant tumor arising from the epithelium of the bile duct, which involves the common hepatic duct, the left and right hepatic ducts, and the confluence areas of these bile ducts. HCCA has an insidious onset and most patients are in the advanced stage when jaundice is observed, and therefore it is considered a difficult issue in the field of surgery. Radical resection is the optimal method for the treatment of HCCA and has great influence on recurrence rate of tumor and patients’ survival time. Therefore, preoperative evaluation of tumor resectability is an important step of HCCA treatment. At present, there are eight main methods for HCCA staging and typing, i.e., Bismuth-Corlette classification system, modified T staging system, TNM staging system, JSBS staging system, Gazzaniga staging system, International Cholangiocarcinoma Group staging system, Mayo staging system, and Blechacz staging system. Each staging and typing system has its own advantages and disadvantages in clinical practice. This article reviews the staging and typing methods for HCCA, with a focus on the clinical value of each staging and typing system in preoperative evaluation of radical resection and prognosis.

miR-223-3p regulates proliferation and apoptosis of hepatocellular carcinoma SMMC-7721 cells by targeting RAC1 / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effects of miR-223-3p on the proliferation and apoptosis of hepatocellular carcinoma (HCC) cells by regulating Ras-related C3 botulinum toxin substrate 1 (RAC1) and its possible mechanism. Methods: Thirty pairs of HCC and corresponding para-cancer tissues resected in Jilin Central Hospital from August 2016 to August 2018 were collected for this study; in addition, human HCC cell lines SMMC-7721, BEL-7402, HepG2 and human normal hepatocyte QSG-7701 were also collected. The expression level of miR-223-3p in HCC tissue and cell lines was detected by qPCR. miR-223-3p mimics, miR-223-3p inhibitor and siRAC1 were transfected into SMMC-7221 cells, respectively. CCK-8 assay, Colony formation assay and Annexin V-FITC/PI staining Flow cytometry were used to detect the proliferation, clone formation and apoptosis of SMMC-7721 cells, respectively. The relationship between miR-223-3p and RAC1 was confirmed by Dual luciferase reporter gene assay. The protein level of RAC1 in SMMC-7721 cells was detected by Western blotting. Results: The expression of miR-223-3p in HCC tissues was significantly lower than that in paracaner tissues (P<0.01), and had significant correlation with pathological characteristics, such as tumor size, TNM stage, EdmondsonSteiner grade (all P<0.05 or P<0.01). miR-223-3p expression in HCC cell lines was significantly lower than that in QSG-7701 cells with the lowest expression in SMMC-7721 cells. Dual luciferase reporter gene assay confirmed that RAC1 was a target gene of miR-223-3p, and miR-223-3p negatively regulated RAC1 expression. Over-expression of miR-223-3p significantly inhibited the proliferation and colony formation (P<0.05 or P<0.01) of SMMC-7721 cells and promoted cell apoptosis (P<0.01). Contrarily, knockdown of miR-223-3p reversed the inhibitory effect of miR-223-3p mimics on cells. Conclusion: miR-223-3p over-expression inhibits proliferation and colony formation and promotes apoptosis of HCC cells, the mechanism of which may be related with its targeted down-regulation of RAC1.

Research progress in process and mechanisms of embryo implantation / 生理学报

The onset of tight connection between embryo and uterine endometrium terms "embryo implantation", the beginning and a key step of mammalian pregnancy. Defective implantation leads to failure of pregnancy and infertility. In recent years, along with the technological advance, researches on embryo implantation have achieved great advances. This paper reviews the key research achievements that have been reached in the last decade in the field of embryo implantation, focusing on the changes, roles, and underlying mechanisms of both luminal and glandular epithelia during implantation process, as well as their interactions with embryo trophoblast cells and endometrial stromal cells.

The mechanism underlying pathogenesis of Parkinson's disease and the progress in its therapeutics / 医学研究生学报

Parkinson′s disease (PD) is a neurodegenerative disorder, characterized by progressive loss of dopaminergic neurons in the substantia nigra. The clinical manifestations of PD include motor and non-motor symptoms. There are around 5 millions PD patients in China, accounting for half of the total PD cases in the world. The mechanisms underlying PD pathogenesis remain to be elucidated, but it is widely accepted that abnormal protein aggregation, mitochondria dysfunction, and oxidative stress are the key causative factors of PD. The regular treatment for PD includes medication of which L-dopa is the most commonly prescribed, surgical treatment, and physiotherapy. In recent year, with the progress in biotechnology and understanding of mechanisms underlying PD, novel therapeutics such as cell replacement, gene therapy, deep brain stimulation emerge, which substantially benefit the treatment of PD. In present review, based on the molecular mechanisms underlying PD pathogenesis, we summarized and discussed current therapeutic strategy on PD.

Evaluation of different staging methods on radical resection rate and prognosis of patients with hilar cholangiocarcinoma / 中华肝胆外科杂志

Objective@#To compare the Bismuth-Corlette typing, modified T-staging and Mayo staging system in predicting the radical resection rates and prognosis of patients with hilar cholangiocarcinoma (HCC).@*Methods@#The clinical data of 138 patients with hilar cholangiocarcinoma treated in the First Bethune Hospital of Jilin University were retrospectively analyzed. Three different staging methods were used.@*Results@#With increase in the classification level of the Bismuth-Corlette classification, the radical resection rate did not significantly decrease (P>0.05). The radical resection rates of stage T1, T2 and T3 in the modified T-staging system were 60.0% (27/45), 36.0% (10/28) and 14.0% (9/65) respectively (all P<0.05). The radical resection rates of patients in the stages I, II, III, IV of the Mayo Staging System were 86.0% (12/14), 50.0% (14/28), 29.0% (19/66) and 3/0% (1/30) respectively (all P<0.05). The overall survival time were no significant differences between the different Bismuth-Corlette and the modified T-staging system patients (P>0.05). However, there were significant differences among the survival rates in the various tumor staging levels using the Mayo Staging System.@*Conclusions@#The modified T-staging system and the Mayo staging system were more accurate than the Bismuth-Corlette typing system in predicting radical resection rates in patients with hilar cholangiocarcinoma. The Mayo staging system was superior to the Bismuth-Corlette typing system and the modified T-staging system in predicting prognosis of patients with hilar cholangiocarcinoma.

Effect on sperm quality of asthenospermia and oligospermia treated with grain-moxibustion combined with medicine therapy / 中国针灸

OBJECTIVE@#To observe the clinical effect of grain-moxibustion combined with medicine therapy for asthenospermia and oligospermia.@*METHODS@#A tatal of 60 patients were randomized into an observation group (30 cases) and a control group (30 cases) according to 1︰1 ratio. In the control group, vitamin E capsules were taken orally one capsule each time, twice a day, and pills 6 g each time, three times a day for a total of 3 months. In the observation group, grain-moxibustion was applied at Guanyuan (CV 4)，Shenshu (BL 23) and Zusanli (ST 36) based on the control group, once a week for 3 months, with a total of 12 times. The sperm concentration and sperm progressive motility were measured by automatic sperm quality analysis system in the two groups, and the clinical effects were compared. Sperm DNA fragmentation index (DFI) in the observation group was measured by sperm nucleus chromosome structure assay (SCSA).@*RESULTS@#①The sperm concentrations and sperm progressive motilities after 1-month, 2-month and 3-month of treatment were increased compared with those before treatment in the two groups (<0.01), and they were increased with time. In the two groups, 2-month and 1-month of treatment, 3-month and 2-month of treatment were compared, the sperm concentrations and sperm progressive motilities were significantly increased (<0.01). The sperm concentrations after 1-month, 2-month and 3-month of treatment in the observation group were higher than those in the control group (<0.01), the sperm progressive motility after 3-month of treatment in the observation group was higher than that in the control group (<0.05). ②After 3-month of treatment，the DFI in the observation group was significantly reduced compared with that before treatment (<0.01). ③The total effective rate in the observation group after 3-month of treatment was 86.7% (26/30), which was superior to 63.3% (19/30) in the control group (<0.05).@*CONCLUSION@#Grain-moxibustion combined with medicine therapy can improve sperm concentration and sperm progressive motility, enhance the integrity of sperm DNA.

Eicosane, pentadecane and palmitic acid: The effects inin vitro wound healing studies

Objective:To examine the wound healing properties of eicosane, pentadecane and palmitic acid by evaluating in term of anti-microbial, anti-inflammatory, proliferation, migration and collagen synthesis.Methods: Anti-microbial activities of Staphylococcus aureus,Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa were evaluated by carrying out disk diffusion and agar well diffusion methods. Growth rate of tested bacteria was also evaluated for 8 h in conjunction with the sample drugs. Besides, U937 cell lines were used as model study for real-time mRNA genes expression studies of TNF-α and IL-12 under the treatment. Proliferation, migration and collagen content synthesis were carried out on human dermal fibroblast.Results:None of the sample drugs possessed significant inhibition of bacteria tested in this study both in disk diffusion and agar well diffusion methods. In contrary, significantly low expressed mRNA gene expression levels of TNF-α and IL-12 were found under the treatment of respective drugs. Meanwhile in proliferation, migration and hydroxyproline content analysis, all the sample drugs showed no significant positive stimulation.Conclusions:This study therefore explains that apart from their potential in downregulating pro-inflammatory cytokines, these three compounds which were examined individually may not be good candidates in promoting wound healing.

Paeoniflorin Promotes Angiogenesis in A Vascular Insufficiency Model of Zebrafish in vivo and in Human Umbilical Vein Endothelial Cells in vitro / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the pro-angiogenic effects of paeoniflorin (PF) in a vascular insufficiency model of zebrafish and in human umbilical vein endothelial cells (HUVECs).</p><p><b>METHODS</b>In vivo, the pro-angiogenic effects of PF were tested in a vascular insufficiency model in the Tg(fli-1:EGFP)y1 transgenic zebrafish. The 24 h post fertilization (hpf) embryos were pretreated with vascular endothelial growth factor (VEGF) receptor tyrosine kinase inhibitor II (VRI) for 3 h to establish the vascular insufficiency model and then post-treated with PF for 24 h. The formation of intersegmental vessels (ISVs) was observed with a fluorescence microscope. The mRNA expression of fms-like tyrosine kinase-1 (flt-1), kinase insert domain receptor (kdr), kinase insert domain receptor like (kdrl) and von Willebrand factor (vWF) were analyzed by real-time polymerase chain reaction (PCR). In vitro, the pro-angiogenic effects of PF were observed in HUVECs in which cell proliferation, migration and tube formation were assessed.</p><p><b>RESULTS</b>PF (6.25-100 μmol/L) could rescue VRI-induced blood vessel loss in zebrafish and PF (25-100 μmol/L), thereby restoring the mRNA expressions of flt-1, kdr, kdrl and vWF, which were down-regulated by VRI treatment. In addition, PF (0.001-0.03 μmol/L) could promote the proliferation of HUVECs while PF stimulated HUVECs migration at 1.0-10 μmol/L and tube formation at 0.3 μmol/L.</p><p><b>CONCLUSION</b>PF could promote angiogenesis in a vascular insufficiency model of zebrafish in vivo and in HUVECs in vitro.</p>

Effects of sex on chronic pancreatitis induced by L-arginine in Kunming mice / 中国病理生理杂志

AIM:To explore the effect of sex on the formation of chronic pancreatitis(CP)by comparing the differences in L-arginine-induced CP model between male and female mice.METHODS:Male(n=42)and female(n=42)Kunming mice were randomly divided into 4 groups:female control group, female CP group, male control group and male CP group(n=18 in each control group, n =6 at each time point; n=24 in each CP group, n=8 at each time point).The mice in CP groups were intraperitoneally injected with 20%L-arginine(3 g/kg,twice/d,1 d/week).After modeling for 2 weeks,4 weeks and 6 weeks,the mice were anesthetized and sacrificed.The morphological changes and the degree of fibrosis in the pancreas were observed by HE and Masson staining.The positive expression rate of F4/80 in the pancreatic tissues was observed by the method of immunohistochemistry.The mRNA expression of interleukin-6(IL-6), α-smooth muscle actin(α-SMA)and fibronectin(FN)in the pancreas were detected by real-time PCR.The protein of pancreas was extracted to detect the expression of α-SMA and FN by Western blot.RESULTS:At 2 weeks,4 weeks and 6 weeks after intraperitoneal injection of L-arginine, the pancreatic tissues were obviously injured and exhibited different degrees of fibrosis in female and male CP groups.At the same time,there were significant differences in the degree of pan-creatic injury between male and female mice,and the degree of pancreatic lesion in the male mice was significantly more severe.The positive rate of F4/80 in the pancreas of male CP mice was significantly higher than that in female CP group at the same time point after modeling.At every time point, the mRNA expression of IL-6 in the pancreas was increased in both female and male CP groups,but that in male CP group was higher(P<0.05).The fibrosis indexes,α-SMA and FN, were both highly expressed at mRNA and protein levels after modeling, but compared with the female group, the time of positive expression in male mice was earlier and the expression level was higher(P<0.05).CONCLUSION: The CP model is successfully established by intraperitoneal injection of 20%L-arginine,and a difference in the degree of pathologic alteration in pancreas between male and female mice exists.CP is more effectively induced by L-arginine in male mice,and the degree of fibrosis is more pronounced.The reason may be related to the sensitivity of male mice to L-arginine,causing more serious inflammatory response.Therefore,male mice are more suitable for establishing CP animal model.

Mechanisms of Fufang Danshen tablets on treating heart failure via regulating AT₁-mediated PLA2-COX2/5-LOX metabolic pathway / 中国中药杂志

Myocardial fibrosis (MF) is an important pathological change involved in the progress from myocardial infarction (MI) to heart failure(HF). Metabolic disorder of arachidonic acid (AA) in cardiomyocytes plays an important role in process of MF. Fufang Danshen tablets is a traditional Chinese medicine (TCM), which showed significant effect on coronary heart diseases and anti-MF. However, the underlying mechanism of anti-MF remains unclear. In this study, HF animal model of myocardial infarction was established by ligation of left anterior descending coronary artery. The heart function of rats in each group was evaluated by echocardiography and hemodynamic measurement. Histological examination, TUNEL and Western blot were used to detect the levels of MF and proteins related to AA metabolism. As a result, MI significantly decreased the levels of ejection fraction (EF), ejection fraction (FS) and left ventricular systolic pressure (LVSP), and these decreases were significantly improved by the treatment of Fufang Danshen tablets. Besides, Fufang Danshen tablets treatment down-regulated the levels of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) in serum. HE, Masson and TUNEL staining results showed that Fufang Danshen tablets treatment could inhibit the inflammatory cells infiltration and attenuate the fibrosis and apoptosis to exert cardioprotective effect. Western blot indicated that Fufang Danshen tablets treatment down-regulated the expressions of AT₁, MMP2, MM9, while up-regulated the expression of AT₂ to inhibit MF. Further mechanism study indicated that Fufang Danshen tablets inhibited MF by down-regulated the expressions of AA metabolism, such as PLA2, P450, COX2 and 5-LOX. In summary, Fufang Danshen tablets can effectively inhibit MF in the ischemic area after MI in rats. The mechanism is related to the regulation of AT₁-mediated PLA2-COX2 metabolic pathway.

Effect of Ginkgo biloba extract (EGb50) on mitochondrial function in SH-SY5Y cells after hypoxia/reoxygenation injury / 中国中药杂志

Ischemic cerebrovascular disease and cerebral ischemia/reperfusion injury threaten the health of human being. We studied the protective effect of Ginkgo biloba extract 50 (EGb50) on the mitochondrial function in SH-SY5Y cells after hypoxia/reoxygenation (H/R) injury and explored its mechanisms, so as to provide new ideas for studies on the treatment for ischemic cerebrovascular disease. We established the H/R injury model in SH-SY5Y cells after administrating EGb50. Subsequently, the mitochondrial membrane potential and the concentration of intracellular Ca²⁺ were measured by flow cytometer. The levels of optic atrophy1 (Opa1) and dynamin-like protein 1 (Drp1) were evaluated by immunofluorescence and western blot. The results showed that the mitochondrial membrane potential was decreased and the level of intracellular Ca²⁺ was increased after H/R injury. Moreover, the expression of mitochondrial fusion protein Opa1 was decreased, while the expression of mitochondrial fission protein Drp1 was increased. However, EGb50 significantly increased the mitochondrial membrane potential and suppressed the level of intracellular Ca²⁺. In addition, EGb50 increased the expression of Opa1 and decreased the expression of Drp1. The results demonstrated that EGb50 has a neuroprotective effect on SH-SY5Y cells after H/R injury, and could improve the energy metabolism and mitochondrial function. The underlying mechanisms may be associated with the regulation of mitochondrial fusion and fission, which provided data support for the treatment of ischemic cerebrovascular disease with EGb50.

Fabrication of Isoniazid/Rifampicin/Poly L-lactic Acid Donut-shaped Implants via Three Dimensional Printing Technique / 中国医学科学院学报

Objective To investigate the possibility of manufacturing dual-drug loaded isoniazid/rifampicin/poly L-lactic acid (PLLA) implant with donut-shaped structure via three-dimensional (3D) printing technique and study the drug release characteristic and biocompatibility of the implant in vitro.Methods PLLA was crushed into particles with diameters around 75-100 μm.Isoniazid and rifampicin bulk drugs were dissolved into the organic dissolvent respectively to be the binding liquid.The 3D printing machine fabricated the donut-shaped implant via binding the PLLA powder layer by layer.Dynamic socking method was used to study the in vitro release characteristics,and cell culture experiment was used to test the cytocompatibility of the implant.Results PLLA slow-release implants were made by using the PLLA powder as matrix and isoniazid/rifampicin organic solvent as binding liquid through 3D printing.The drugs in the implants distributed in nest under electron microscope.The concentrations of both drugs were still higher than the lowest effective bacteriostasis concentration after release for 32 days.Cytotoxicity and direct contact tests indicated that the implants had rare cytotoxicity and favorable biocompatibility. Conclusion The donut-shaped implants can be successfully fabricated using the 3D printing method,which offers a new method for the manufacturing of topical slow-release anti-tuberculosis drugs.