Objective To investigate the effect of evodiamine(EVO)on retinal injury in diabetic rats by regulating the cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)signaling pathway.Methods Totally 96 Sprague-Dawley rats(192 eyes)were divided into the negative control(NC)group,Model group,low-dose EVO(EVO-L)group,medium-dose EVO(EVO-M)group,high-dose EVO(EVO-H)group,calcium dobesilate(CD)group,SQ22536 group and EVO-H+SQ22536 group,with 12 rats in each group.Rats in the NC group were intraperitoneally injected with physiological saline instead of streptozotocin,and diabetic retinopathy models were established in all other groups.After successful mod-eling,the drug was administered once a day for 4 weeks.The blood glucose level of rats in each group was measured by blood glucose meter;HE staining was applied to detect the pathological changes of the retina of rats;TUNEL staining was adopted to detect the apoptosis of ganglion cells in the retina of rats;the levels of superoxide dismutase(SOD),malondial-dehyde(MDA),tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and cAMP in the retina of rats were detected;West-em blot was used to detect the protein expressions of Bcl-2 associated X protein(Bax),P53 and phosphorylated protein ki-nase A(p-PKA)in the retina of rats.Results Compared with the NC group,the pathological injury of the retina in the Model group was more serious;the blood glucose,apoptosis rate of retinal ganglion cells,MDA,TNF-α,IL-6,Bax and P53 protein expressions increased,and the SOD,cAMP and p-PKA/PKA protein expression decreased,with statistically significant differences(all P＜0.05).Compared with the Model group,the pathological injury of the retina was relieved,the blood glucose,apoptosis rate of retinal ganglion cells,MDA,TNF-α,IL-6,Bax and P53 protein expressions decreased,and the SOD,cAMP and p-PKA/PKA protein expression increased in the EVO-L group,EVO-M group,EVO-H group and CD group,with statistically significant differences(all P＜0.05).Compared with the Model group,the retinal tissue of the SQ22536 group was severely damaged,the blood glucose,apoptosis rate of retinal ganglion cells,MDA,TNF-α,IL-6,Bax and P53 protein expressions increased,and the SOD,cAMP,p-PKA/PKA protein expression decreased,with statistically significant differences(all P＜0.05).Compared with the EVO-H group,the EVO-H+SQ22536 group showed more serious pathological injury of retinal tissue,increased blood glucose,apoptosis rate of retinal ganglion cells,MDA,TNF-α,IL-6,Bax and P53 protein expressions,and decreased SOD,cAMP and p-PKA/PKA protein expression,and the differences were statistically significant(all P＜0.05).Conclusion EVO may alleviate retinal injury in diabetic rats by activating the cAMP/PKA signaling pathway.

Objective:To explore the short-term efficacy and safety of intra-arterial thrombolysis (IAT) in the treatment of retinal artery occlusion (RAO) with the assistance of the rescue green channel in the eye stroke center.Methods:A prospective, interventional, single-center study. Thirty-eight eyes from 38 RAO patients who received IAT treatment in Guangdong Provincial People’s Hospital were enrolled. All the patients were rescued via the green channel in our eye stroke center. Data from comprehensive ocular examinations including best-corrected visual acuity (BCVA) and optical coherence tomography angiography (OCTA) were collected. BCVA was measured with Snellen chart and converted to the logarithmic minimum angle of resolution (logMAR) unit for statistical analysis. RTVue XR OCTA was used to measure vascular densities (VD) of the superficial capillary plexus (SCP), deep capillary plexus (DCP) and radial peripapillary capillary (RPC), and central retinal thickness (CRT). All RAO eyes attempted IAT treatment and 34 eyes were successful. Four eyes failed to complete IAT because of the occlusion of internal or common carotid arteries on the same side with the RAO eyes. Ocular examinations in post-operative 1-3 days were performed with the same devices and methods as those before surgery. Parameters measured before and after surgery include BCVA, VD of SCP, DCP, RPC, and CRT. Data of the green channel collected include the time intervals from onset of RAO to first presentation in local hospitals, and from onset of RAO to our eye stroke center. Comparisons of VD and CRT between the RAO eyes and contralateral healthy eyes were performed with independent samples Mann-Whitney U test; comparisons of VD and CRT in RAO eyes before and after IAT surgery were performed with paired samples Wilcoxon Rank Sum test. Results:Among the 34 RAO patients who had successful IAT surgery, 18 (52.9%, 18/34) were males and 16 (47.1%, 16/34) were females; the mean age was (51.0±12.9) years old. There were 30 and 4 eyes diagnosed as central RAO and branch RAO respectively. The logMAR BCVA before and after IAT surgery was 2.52±0.61 and 2.18±0.85 respectively, and the difference was statistically significant ( Z=-3.453, P=0.002). Before surgery, VD of SCP, DCP and RPC were significantly decreased and CRT was significantly increased in the affected eye compared with the contralateral healthy eyes, with the statistical significance ( P<0.001). Compared with those before surgery, the VD of SCP and DCP were significantly improved after surgery ( Z=-2.523, -2.427; P=0.010, 0.014), while there was no difference in VD of RPC and CRT ( Z=-1.448, -1.454; P=0.150, 0.159). The time interval between onset of RAO and first visit to the hospital was (6.56±6.73) hours; the time interval between onset of RAO and the arrival at our hospital was (24.11±19.90) hours. No cerebral stroke was observed in the early postoperative period and no cerebrocardiovascular events were observed later. he time interval between onset of RAO and the arrival at our hospital was (24.11±19.90) hours. No cerebral stroke was observed in the early postoperative period and no cerebrocardiovascular events were observed later. Conclusions:The short-term efficacy and safety of IAT in the treatment of RAO were satisfactory. The rescue time window might be prolonged.

Objective:To observe alterations of macular outer retinal reflectivity (ORR) and the associations with macular vessel density in patients with nonproliferative diabetic retinopathy (NPDR).Methods:A retrospective cross-sectional study. From August 2021 to March 2022, a total of 63 NPDR patients with 63 eyes (NPDR group) diagnosed by Department of Ophthalmology of Guangdong Provincial People'sHospital were included in the study. There were 39 males with 39 eyes and 24 females with 24 eyes. Age was 60 (52, 68) years. A total of 66 eyes of 66 healthy volunteers matching age and sex were selected as the control group. Among them, 40 men had 40 eyes and 26 women had 26 eyes. Age was 58 (52, 67) years. Optical coherence tomography (OCT) and OCT angiography (OCTA) were performed in all affected eyes. Image J software was used to calculate ORR, including the optical density of ellipsoid zone (EZ), photoreceptor outer segment (OS), photoreceptor inner segment (IS) and outer nuclear layer (ONL) by OCT examination. The sampling sites were horizontal and vertical scanning of the fovea of the macula on 500 μm (nasal 500, temporal 500, superior 500, inferior 500), 1 000 μm (nasal 1 000, temporal 1 000, superior 1 000, inferior 1 000) and 2 000 μm (nasal 2 000, temporal 2 000, superior 2 000, inferior 2 000). The software automatically divided the retina within 6 mm of the macular fovea into the fovea with a diameter of 1 mm, the parafovea with a diameter of 1-3 mm, and the perifovea with a diameter of 3-6 mm by macular OCTA examination. The blood density of superficial capillary plexus and deep capillary plexus in different zones in the macular area were measured by the built-in software of the device. Spearman correlation analysis was used to analyze the correlation between ORR and blood flow density. Results:Compared with the control group, retinal reflectivity of EZ in NPDR group was significantly decreased at other sites except the fovea, retinal reflectivity of OS was significantly decreased at nasal 2 000, temporal 2 000, superior 2 000 and superior 1 000; retinal reflectivity of IS was significantly decreased at superior 1 000, superior 500 and inferior 500. The retinal reflectivity of ONL in macular fovea was significantly decreased, and the differences were statistically significant ( P<0.05). The ORR was positively correlated with blood flow density, and the correlation coefficient in NPDR group was lower than that in control group. The results of multifactor linear regression analysis showed that the superior and temporal ORR were correlated with blood flow density ( P<0.05). Conclusions:Compared with the control group, ORR is reduced and less correlated with vessel density in NPDR patients. ORR is more affected by retinal blood flow density in temporal and superior parts.

【Objective】 To investigate the main causes of blood donor deferral in domestic blood center. 【Methods】 The causes of donor deferral were classified into 12 categories as previous medical history, drug use, alcohol consumption, menstrual period, underweight, abnormal blood pressure, abnormal body temperature, abnormal hemoglobin (Hb), lipemic blood, positive hepatitis B surface antigen (HBsAg), elevated alanine aminotransferase (ALT) and others according to the comparison indicators of Asia-Pacific Blood Network (APBN) and the national standard Blood Donor Health Examination Requirements. The relevant data of the top 3 causes of donor deferral, voluntarily reported by the members of Practice Comparison Working Group of China’s Mainland Blood Collection and Supply Institutions from 2014 to 2019, were collected and a histogram was generated. 【Results】 The median donor deferral rate of 20 domestic blood centers from 2014 to 2019 was 12.14%, with the lowest at 0.18% and highest at 32.32%, respectively. The top three causes for donor deferral were elevated ALT, abnormal Hb and abnormal blood pressure in year 2014, 2015, 2018 and 2019; elevated ALT, lipemic blood and abnormal blood pressure in 2016; elevated ALT, abnormal Hb, and lipemic blood in 2017. 【Conclusion】 The main causes of donor deferral were elevated ALT, abnormal Hb, abnormal blood pressure and lipemic blood.

Objective To observe Rongxin Pills in the treatment of viral myocarditis in children (deficiency of both qi and yin and heart meridian stasis syndrome) and the effectiveness and clinical application of safety.Methods Viral myocarditis patients (280 cases,deficiency of both qi and yin and heart meridian stasis syndrome),according to 3:1 ratio as the test group (n =21 0) and control group (n =70).The test group took orally Rongxin pills each time 4.5～9 g,3 times daily;the control group oral coenzyme Q10 capsule each time 10 ～ 20 mg,twice daily.The course of treatment was 28 d.The experiment was carried out with the random and double blind method.The symptoms of myocarditis,integrated and electrocardiogram,echocardiography,myocardial enzymes,as well as the efficacy of traditional Chinese medicine and improvement of the effect of the disease were observed.Results The results of FAS (PPS) analysis showed that 28 d after treatment,the symptom score and mean of experimental group and control group were 5.975 (6.000) and 4.721 (4.788).The syndromes of the total effective rates were 91.62％ (90.59％) and 70.59％ (71.21％),curative effect the total effective rates were 90.14％ (92.08％) and 72.06％ (72.73％).The total effective rate of experimental group was higher than that of the control group,the difference was statistically significant.In this experiment,three cases of clinical adverse events were reported,which were not related to the experimental drug.It also not belongs to adverse drug reactions.Conclusion Rongxin Pill in the treatment of viral myocarditis in children (deficiency of both qi and yin and heart meridian stasis syndrome) is more effective than coenzyme Q 10 capsule,and there was no indication of higher risk of clinical application.

AIM:To explore the effects of different intensity of warfarin on patients with coronary atherosclerotic heart disease complicated with atrial fibrillation.METHODS:One hundred and seven cases of coronary heart disease complicated with atrial fibrillation patients in our hospital were selected and were randomly divided into experimental group (n =54) and control group (n =53) by random number table.The experimental group (low intensity) received initial amount of warfarin for 1.25 mg/d,and NR monitoring 24 h after treatment,if INR＜ 1.4,3-5 mg/d increased by 0.5-1.0 mg/d,monitoring one time per week,INR maintained within 1.4-2.0.INR of the control group (medium intensity) maintained within 2.0-2.6.One month monitoring after INR stabilized.All patients were treated for about 4 weeks,and warfarin was maintained at a dose of 1.25-7.5 mg/d.The primary and secondary end points and bleeding events were observed and compared after 2 years of treatment.RESULTS:The INR of the experimental group and the control group were 1.71 ± 0.38,2.36 ± 0.35,respectively.The ratio of total bleeding event of the experimental group and the control group was 22.2％ and 41.5％,respectively.Those of the experimental group were lower than those of the control group,and the difference was all statistically significant (P＜ 0.05).CONCLUSION:The efficacy of low-intensity warfarin in the treatment of coronary atherosclerotic heart disease complicated with atrial fibrillation is comparable to that of moderate-intensity warfarin therapy,but low-intensity warfarin shows better saftey.

Objective:To study the antitumor activity and immunological mechanisms of rBCG including GM-CSF and EB virus LMP2A gene fusion.Methods: Animal models of EB virus-positive tumors was built.The formation time of tumors in mice,survival time,tumor weight was analyzed to detect rBCG anti-tumor activity;ELISA method was used to detect the specific antibodies which was produced in the mice stimulated by rBCG,specific CTL killing effect was detected by lactic dehydrogenase assay,ELISPOT was used to assay the secretion of IFN-γand flow cytometry, HE staining of tumor tissue was used to detected lymphocyte infiltration in mice immunized with recombinant BCG.Statistical methods were used for rBCG immunization effect preliminary analysis and evaluation.Results:Comparing to other control,tumor formation time was significantly delayed and tumor growth was slow, survival time of mice prolonged .ELISA test results showed that the rBCG immunization groups of mice could produce specific IgG antibodies of GM-CSF and LMP2A.Specific CTL activity was detected in mice immunized with rBCG.IFN-γsecretion was detected by ELISPOT method, flow cytometry and morphological observation detected tumor tissue infiltration of lymphocytes growth in mice immunized with rBCG.Conclusion:The rBCG induced a humoral and cellular immune responses and induced C57BL/6 mice to produce a strong anti-tumor effect and the EB virus-positive tumor cells was significantly inhibited.

Objective To construct mutant strains of methicillin resistant Staphylococcus epidermi-dis (MRSE) with psm-mec gene deletion and to investigate the function of psm-mec gene.Methods The drug sensitivity test and DNA sequence analysis were performed to screen out the tetracycline and chloram -phenicol sensitive clinical strains of MRSE , whose upstream and downstream sequences of psm-mec gene were identical to those of the Staphylococcus epidermidis reference strain RP62A.The recombinant plasmid pBT2-Δpsm-mec was constructed by using the fusion PCR and a temperature sensitive shuttle plasmid .After being identified , the plasmid was transformed into the Staphylococcus aureus RN4220 strain by electropora-tion, and then transformed into the selected clinical isolates of MRSE .The mutant strains of MRSE with psm-mec deletion were screened out and identified after homologous recombination .The differences in biofilm formation between the mutant and wild-type strains were analyzed for further elucidation the relationships be-tween the psm-mec gene and biofilm formation in MRSE strains .Results Three clinical MRSE isolates for the construction of mutant strains with psm-mec gene deletion were screened out and identified by using drug sensitivity test and sequence alignment analysis .The mutants constructed via homogenous recombination were screened out and identified .Compared with the corresponding wild-type strains, the three mutants with psm-mec gene deletion showed significantly decreased ability of biofilm formation , demonstrating that the psm-mec genes strains induced the biofilm formation of MRSE .Conclusion The Δpsm-mec mutant strains were successfully constructed .The psm-mec gene played an important role in the biofilm formation of Staphy-lococcus epidermdis.

AIM:To investigate the effects of atorvastatin reloading in pre-percutaneous coronary intervention ( PCI) period on endothelial progenitor cell ( EPC) count and inflammatory cytokine expression in the stable angina pectoris patients who had previously received long-term statin treatment.METHODS:The patients with stable angina pectoris that had received long-term statin therapy and planned to accept PCI were randomized into 3 groups:80 mg atorvastatin 12 h and 40 mg 2 h before coronary angioplasty (80 mg reloading), pre-operatively with 40 mg/d atorvastatin for 7 d (40 mg re-loading) , and without atorvastatin reloading ( no reloading ) .CD45 -/CD133+/CD34 +, CD45 -/CD34 +/KDR+ and CD45 -/CD144 +/KDR+EPCs in 100 μL peripheral blood were determined by flow cytometry 1 h prior to PCI and 1 h, 6 h and 24 h after PCI.The serum concentrations of soluble intercellular adhesion molecule 1 ( sICAM-1) , C-reactive protein ( CRP) and troponin I ( TnI) were analyzed immediately prior to and 24 h after PCI.RESULTS:(1) In 80 mg reloading group, the numbers of circulating CD45 -/CD133 +/CD34 +and CD45 -/CD34 +/KDR+early differentiation stage EPCs 1 h and 6 h after coronary angioplasty was significantly elevated compared with those before PCI (P<0.05).(2) In control group, the serum concentrations of sICAM-1 and CRP 24 h after PCI were significantly elevated ( P<0.05) compared with preoperative values.(3) The rise in serum TnI concentration from pre-to post-operation in 80 mg reloading group was lowerthan that in control group.CONCLUSION: The method of atorvastatin reload before PCI affects the number of EPCs inperi-operative period.High dose of atorvastatin application before PCI triggers early EPC circulation.The serum levels ofpost-operative inflammatory cytokine sICAM-1 as well as CRP are reduced by atorvastatin reloading before PCI.

OBJECTIVETo investigate the effect of hydroxycamptothecin (HCPT) on the proliferation, cell cycle and apoptosis of human lung carcinoma cell line A549.

METHODSThe growth of A549 cells exposed to HCPT was observed by staining with acridine orange/ethidium bromide dye. Agarose gel electrophoresis was performed to detect DNA fragmentation of the apoptotic cells. The cell cycle distribution of the exposed cells was analyzed using flow cytometry, and cell apoptosis was examined with annexin V-FITC/PI staining. RT-PCR was used to investigate Bcl-2 gene expression changes in the exposed cells.

RESULTSAgarose gel electrophoresis of the DNA from HCPT-treated cells showed a DNA ladder, and typical apoptotic appearance of the exposed cells was observed under fluorescence microscope. Treatment of A549 cells with 1 µmol/L HCPT for 24 h resulted in a cell apoptosis rate of 18.11%, significantly higher than the rate in control cells (0.09%, P<0.05). The treatment also caused a significant reduction of Bcl-2 mRNA expression by 70% (P<0.05).

CONCLUSIONHCPT can significantly inhibit the proliferation, induce apoptosis, and down-regulate Bcl-2 gene expression in human lung carcinoma cell line A549, suggesting the involvement of Bcl-2 gene in the inhibitory effect of HCPT on A549 cells.

Camptothecin ; analogs & derivatives ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; drug effects ; Genes, bcl-2 ; Humans ; Transfection