Objective To construct vectors that express phosphatidylinositol-3-kinase, catalytic, beta polypeptide （PIK3cb） shRNA in eukaryon plasmid catalyzed by PI3K in rat, then test their effects on intimal hyperplasia in transplanted vein graft. Methods One hundred and fifty SD rats were randomly divided into six groups （n=25, in each group）: blank (25% Pluronic F-127), shRNA-1, shRNA-2, 1/2 （shRNA-1+shRNA-2）, negative control (pGenesil-1 scramble shRNA) and positive control （wortmannin） group. The jugular vein in rats were interpositioned autologously into the common carotid artery. shRNA and 25% Pluronic F-127 were mixed and coated around the transplanted vein in three PIK3cb shRNA groups. Every 5 samples were removed according to the time point (1, 3, 7, 14 and 28 days after operation), respectively. The thickness of intima and neointima area were calculated and analyzed by computer system. The PCNA expression was detected by Western blot and SP immunohistochemistry. Results The intimal thickness of three PIK3cb shRNA groups were lower than those in the blank group and negative control group on day 3, 7, 14, 28 after operation (P＜0.05); The neointima area in three PIK3cb shRNA groups (except shRNA-2 group on day 3, 7) began to decrease significantly from day one (P＜0.05). The protein expression of PCNA in three PIK3cb shRNA groups on day 3 after operation were decreased compared with blank group and negative group (P＜0.05). The percentage of the PCNA positive cells area in three PIK3cb shRNA groups were significantly lower than those in blank group and negative control group in each time point (Plt;0.05). There were no significant differences between blank and negative control group in different time points (Pgt;0.05). Conclusion The PIK3cb shRNA can effectively inhibit the proliferation of vascular smooth muscle cell, which may provide a new gene therapy for the prevention of vein graft restenosis after bypass grafting.
ObjectiveTo systematically review the efficacy of treatments for β-coronaviruses.MethodsPubMed, EMbase, Web of Science, The Cochrane Library, SinoMed, CNKI and WanFang Data databases were electronically searched to collect randomized controlled trials (RCTs) and non-randomized controlled trials (non-RCTs) of treatments for β-coronaviruses from inception to June 17th, 2020. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies. Meta-analysis was then performed using Stata 14.0 software.ResultsA total of 109 studies invoving 23 210 patients were included. The results of the systematic review showed that compared with standard of care, corticosteroids could reduce mortality and increase cure rate for COVID-19. However, chloroquine could decrease cure rate. In severe acute respiratory syndrome (SARS) patients, corticosteroids could decrease the cure rate. In Middle East respiratory syndrome (MERS) patients, ribavirin/interferon/both drugs showed higher mortality.ConclusionsThe currently limited evidence shows that corticosteroids may be effective to COVID-19 patients while having limited effects on SARS patients. Hydroxychloroquine or chloroquine may have negative effects on COVID-19 patients. Ribavirin/interferon may be harmful to MERS patients. Due to limited quality and quantity of the included studies, more high quality studies are needed to verify the above conclusions.
【Abstract】ObjectiveTo explore the effects of RECK gene on the biological behaviors of hepatocellular (HepG2). MethodsThe RECK cDNA was transfected to HepG2 with lipofectamine 2000. Detected its protein expressions with Western blot before and after transfection, analyzed the effects of RECK on MMP-9 activity using gelatin zymography, observed the effects on proliferation ability by MTT assay and plate clone formation assay, compare the changes of invasion ability by cell adhesion assay and in vitro invasion experiment. Results RECK protein was expressed steadily in transfected HepG2 cells and the amount of activated MMP-9 were decreased significantly. Their proliferation abilities weren’t different before and after transfection but their invasion abilities decreased sharply. ConclusionRECK gene can transfect HepG2 cells by liposome efficiently. It can inhibit the activity of MMP-9 and the invasion ability of HepG2.
ObjectiveTo evaluate effect of RAS gene mutation after liver metastasis resection on overall survival (OS) and disease-free survival (DFS) for patients with colorectal cancer combined with liver metastasis. MethodsA comprehensive and systematic literature search in the PubMed and other databases was conducted, with the final search ending on January 5, 2022. The impact of RAS gene mutation after liver metastasis resection on survival of patients with colorectal cancer combined with liver metastasis was analyzed by the Stata 12.0 software and Review Manager version 5.3 software, meanwhile which were analyzed according to subgroups, including study type (retrospective and prospective studies), region (Asian and European), and number of RAS gene mutation sites (>2 and ≤2). ResultsA total of 26 studies with 13 356 patients were included. The integrated analysis results showed that the patients with RAS mutations had statistically shorter OS [HR=1.54, 95%CI (1.43, 1.65), P<0.001] and DFS [HR=1.32, 95%CI (1.19, 1.44), P<0.001] as compared with RAS wild-type. Except the 1-year overall survival rate, the 2–5-year overall survival rate and 1–5-year disease-free survival rate of patients with RAS gene mutation were statistically lower than those of patients with RAS wild-type (P<0.05). The results of subgroup analysis showed that no matter retrospective and prospective studies, as well as studies in Asian and European countries, it was found that the OS and DFS for patients with RAS gene mutation were shorter than those of patients with wild-type (P<0.05); At the same time, subgroup analysis of the number of RAS gene mutation sites showed that OS and DFS of patients with number of mutation sites >2 were shortened as compared with ≤2 (P<0.05). ConclusionFrom the overall analysis results, the survival of patients with RAS gene mutation after liver metastasis resection is worse than that of patients with RAS wild-type for patients with colorectal cancer combined with liver metastasis.
ObjectiveTo identify 3 the disease-causing genes and mutations of Leber congenital amaurosis (LCA), and to study the correlation of phenotype and genotype. MethodsA retrospective study. Four LCA patients and seven family members who were diagnosed by eye examination in Ning Xia Eye Hospital of People's Hospital of Ningxia Hui Autonomous Region from January to December 2021 were included in the study. Four patients were from 3 unrelated families. Detailed collection of medical history and family history were received. Related ophthalmologic examination were collected and genomic DNA was extracted from peripheral blood. Whole-exome sequencing method was used for genetic diagnosis. The identified variant was confirmed with Sanger sequencing. Potential pathogenic mutation was analyzed using software and conserved domain analysis and performed co-separated analysis between the family member and the proband. ResultsOf the 4 patients, 1 patient was males and 3 patients were females; the age was from 4 to 18 years. Nystagmus were seen in 3 cases, finger pressing eyes and night blindness was seen in 1 cases; electroretinogram showed 4 cases of extinction or near extinction. The foveal reflection was visible in all eyes, and there was no obvious abnormality in the peripheral retina. One eye had strong reflection signal with raised ellipsoid in macular area; two eyes had weak reflection signal faintly visible between retinal layers; 1 eye had increased blood vessel branches, peripheral retinal non-perfusion area with capillary leakage; annular strong autofluorescence in macular area 4 eyes. No obvious abnormality was found in the phenotypes of family members. Genetic testing showed that the proband of pedigree 1 (Ⅱ-1) was found a homozygous missense mutation in c.640A>T (p.C214S) (M1) of PRPH2 gene. The proband of pedigree 2 (Ⅱ-2) was found compound heterozygous mutation in c.1256G>A(p.R419Q) (M2) and c.1A>C (p.M1L) (M3) of TULP1 gene. The proband 3 (Ⅱ-1) and her sister (Ⅱ-2) were both found compound heterozygous mutation in c.1943T>C (p.L648P) (M4) and c.380C>T (p.P127L) (M5) of GUCY2D gene. The parents and sister (Ⅱ-1) of the proband in family 2 and the parents of the proband in family 3 were all carriers of the corresponding heterozygous variant. M1, M3, M4, M5 were novel mutations and unreported. The genotype and disease phenotype were co-segregated within the family. According to the analysis of pedigree and genetic testing results, all 3 families were autosomal recessive inheritance. The amino acid conservation analysis found that M1, M2, M3, M4, and M5 were highly conserved among species. The results of bioinformatics analysis were all pathogenic variants. ConclusionsPRPH2 gene M1, TULP1 gene M3, and GUCY2D gene M4, M5 were novel mutations and not been reported in the literature and database. This research expanded the gene mutation spectrum of LCA. The patients with LCA have available characterristics, including onset age, varying ocular fundus and severe visual impairment.
ObjectiveTo investigate the feasibil ity of the domestic porous tantalum as scaffold material of bone tissue engineering by observing the expressions of osteogenesis related factors of MG63 cells co-cultured with domestic porous tantalum.
MethodsMG63 cells were cultured with porous tantalum scaffolds (group A), with porous tantalum leaching solution (group B), and with MEM as control group (group C). The cell adhesion of group A was observed on the scaffolds at 3, 5, and 7 days after culture by scanning electron microscopy (SEM); immunohistochemistry and Western blot methods were used to detect the expressions of Runt-related transcri ption factor 2 (Runx-2), osteocalcin (OC), and fibronectin (FN).
ResultsAt 3 days after culture, the cells of group A adhered the surface and pore of the porous tantalum scaffolds, with sparse cell arrangement and less protuberances; at 5 days after culture, adjacent cells connected to be a flat each other, which covered the surface and pore of the scaffold; at 7 days after culture, cells secreted plenty of extracellular matrix, covering most of the material surface. The expressions of Runx-2, OC, and FN were positive in 3 groups; darker staining of the cytoplasm was observed in group A, the expressions were significantly higher in group A than in other 2 groups. The results of immunohistochemistry and Western blot showed that the expressions of Runx-2 and OC were significantly increased in group A when compared with those in groups B and C (P < 0.05), but no significant difference was found between groups B and C (P > 0.05). The expression of FN had no significant difference among 3 groups (P > 0.05).
ConclusionDomestic porous tantalum could promote MG63 cells adhesion and growth, and may promote the expressions of Runx-2 and OC, so it can be used as a scaffold material of bone tissue engineering.
Based on the capacitance coupling principle, we studied a capacitive way of non-contact electrocardiogram (ECG) monitoring, making it possible to obtain ECG on the condition that a patient is habilimented. Conductive fabric with a good electrical conductivity was used as electrodes. The electrodes fixed on a bed sheet is presented in this paper. A capacitance comes into being as long as the body gets close to the surface of electrode, sandwiching the cotton cushion, which acts as dielectric. The surface potential generated by heart is coupled to electrodes through the capacitance. After being processed, the signal is suitable for monitoring. The test results show that 93.5% of R wave could be detected for 9 volunteers and ECG with good signal quality could be acquired for 2 burnt patients. Non-contact ECG is harmless to skin, and it has advantages for those patients to whom stickup electrodes are not suitable. On the other hand, it is convenient to use and good for permanent monitoring.
ObjectiveTo systematically review the efficacy of different stimulation modalities of repetitive transcranial magnetic stimulation (rTMS) combined with SSRI in improving depressed mood after stroke using network meta-analysis. MethodsThe PubMed, EMbase, Cochrane Library, Web of Science, CNKI, VIP, CBM and WanFang Data databases were electronically searched to collect randomized controlled trials (RCTs) related to the objectives from inception to October 1, 2022. Two reviewers independently screened literature, extracted data and assessed the risk of bias of the included studies. Network meta-analysis was then performed by using R 4.2.1software. ResultsA total of 25 RCTs involving 2 152 patients were included. Four types of rTMS stimulation combined with SSRIs were included: high-frequency stimulation of the left dorsolateral prefrontal (l-DLPFC), low-frequency stimulation of l-DLPFC, low-frequency stimulation of the right dorsolateral prefrontal (r-DLPFC), and low-frequency stimulation of the bilateral DLPFC. The results of the network meta-analysis showed that the effect of combining four stimulation methods with SSRI in treating depression was better than that of SSRI alone (P<0.05). Probability sorting results showed that low-frequency stimulated bilateral DLPFC (88.9%) > low-frequency stimulated l-DLPFC (63.1%) > high-frequency stimulation l-DLPFC (57.1%) > low-frequency stimulation r-DLPFC (40.4%). There was no statistically significant difference in the incidence of adverse reactions between the four stimulation methods combined with SSRI and the use of SSRI alone (P>0.05). Conclusion?rTMS combined with SSRIs is better than SSRIs alone in improving depressed mood after stroke. Low-frequency rTMS stimulation of bilateral DLPFC may be the best. Meanwhile, the safety of different stimulation methods is good.
The theoretical foundation of relevant packages of R software for network meta-analysis is mainly based on Bayesian statistical model and a few of them use generalized linear model. Network meta-analysis is performed using GeMTC R package through calling the corresponding rjags package, BRugs package, or R2WinBUGS package (namely, JAGS, OpenBUGS, and WinBUGS software, respectively). Meanwhile, GeMTC R package can generate data storage files for GeMTC software. Techonically, network meta-analysis is performed through calling the software based on Markov Chain Monte Carlo method. In this article, we briefly introduce how to use GeMTC R package to perform network meta-analysis through calling the OpenBUGS software.
Abstract： Objective To observe the expression changes of microRNA 1 （miRNA-1） and microRNA 21（miRNA-21） after ischemic preconditioning （IPC）, ischemic postconditioning （IPO） and remote ischemic preconditioning （RIPC）in an ischemia-reperfusion rat heart model in vitro, as well as the expression of their target protein heat shock protein 70 （HSP70） and programmed cell death 4 （PDCD4）, and evaluate whether miRNA are involved in endogenous cardio-protective mechanism. Methods The Langendorff-perfused Sprague-Dawley rat hearts were randomly assigned into one of the four groups, control group （CON group, n=12）, ischemia preconditioning group （IPC group， n=12）?, ischemia postconditioning group （IPO group， n=12） and remote ischemia preconditioning group （RIPC group，n=12）. Cardiac function was digitalized and analyzed. The expression of HSP70, PDCD4, B-cell lymphoma/leukemia-2 （Bcl-2） and Bax was detected by Western blotting. The expression of miRNA-1 and miRNA-21 was detected by real-time reverse transcriotion-polymerase chain reaction （RT-PCR）. Assessment of cardiac infarct size and myocardial apoptosis was determined using triphenyltetrazolium chloride （TTC） assay and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay （TUNEL） assay respectively. Results The expressions of miRNA-1 and miRNA-21 were up-regulated in IPC group, but the expression of miRNA-1 was down-regulated in RIPC group and IPO group （P＜0.05）. The expressionsof PDCD4, HSP70 and Bax were down-regulated in ‘conditioning’ groups compared with CON group （P＜0.05）. The expression of Bcl-2 was not statistically different among the four groups. The infarct size and the myocardial apoptosis in ‘conditioning’ hearts were significantly decreased compared with CON group （P＜0.05）. Conclusion The expressions of the miRNA-1 and miRNA-21 are different in IPC, RIPC and IPO groups, and their target proteins are not inversely correlated with the miRNAs in all the ‘conditioning’ groups.
