Developing and implementing biosafety standards for pathogenic microbiology laboratories is essential to achieving scientific, efficient, and standardized management and operation. This article analyzes the current standardization construction in biosafety in pathogenic microbiology laboratories domestically and internationally. It proposes a framework for the biosafety standard system of pathogenic microbiology laboratories, which mainly includes four parts: basic standards, management standards, technical standards, and industry applications. It provides a reference for the standardization work of pathogenic microbiology laboratories and helps to standardize the biosafety industry in China.

AIM:To investigate the impact of celastrol intervention on excitatory amino acid transporter 2(EAAT2)and its neuroprotective role in subarachnoid hemorrhage(SAH).METHODS:Western blot analysis was uti-lized to assess the EAAT2 expression level within 72 h after SAH,while glutamate concentration in cortical brain tissues was measured.Computational simulation was employed to explore the binding of celastrol with EAAT2.Seventy SD rats were randomly assigned to sham,model,model+GT949(an EAAT2 agonist),model+dihydrokainic acid(DHK;an EAAT2 inhibitor),and model+celastrol groups.Glutamate concentration in cortical brain tissues was quantified,and brain edema was assessed by dry-wet weight method.Western blot analysis was conducted to evaluate the expression of EAAT2,aquaporin 4 and apoptosis-related proteins(Bax,Bcl-2,caspase-3 and caspase-9),and TUNEL staining was employed to assess the apoptotic cell count in each group.RESULTS:(1)EAAT2 level decreased while glutamate con-centration increased.(2)Celastrol was found to directly bind to EAAT2,enhancing EAAT2 expression and reducing glu-tamate concentration after SAH.(3)Celastrol demonstrated the ability to inhibit brain edema after SAH.(4)Celastrol was effective in reducing neuronal apoptosis after SAH.CONCLUSION:Celastrol has the potential to up-regulate EAAT2 expression,lower glutamate level,mitigate brain edema,and decrease neuronal apoptosis after SAH.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To investigate the influencing factors for celiac lymph node metastasis in thoracic esophageal squamous cell carcinoma (TE-SCC), construct a prediction model of celiac lymph node metastasis in TE-SCC, and stratify the probability of celiac lymph node metastasis.Methods:The retrospective case-control study was conducted. The clinicopathological data of 443 patients with TE-SCC who underwent thoracoscopic and laparoscopic esophagectomy with systematic lymph node dissection in the First Affiliated Hospital of Zhengzhou University between March 2015 and April 2019 were collected. There were 259 males and 184 females, aged from 41 to 81 years, with a median age of 64 years. The nomogram prediction model was constructed based on the results of multivariate analysis of influencing factors for celiac lymph node metastasis in TE-SCC, of which calibration curve and decision curve were drawed. The predictive performance was evaluated using the concordance index. The score for celiac lymph node metastasis in TE-SCC predicted by nomogram model was used for further recursive partitioning analysis, and patients were stratified into risk subgroups using the decision-making tree model. Observation indicators: (1) celiac lymph node metastasis in TE-SCC; (2) analysis of influencing factors for celiac lymph node metastasis in TE-SCC; (3) construction of nomogram prediction model of celiac lymph node metastasis in TE-SCC; (4) construction of decision-making tree model of celiac lymph node metastasis in TE-SCC and risk subgroup analysis of celiac lymph node metastasis probability. Measurement data with skewed distribution were represented as M (range). Count data were represented as absolute numbers and percentages, and comparison between groups was analyzed using the chi-square test. Comparison of ordinal data between groups was analyzed using the nonparametric rank sum test. Multivariate analysis was performed using the Logistic regression model. Based on Logistic regression model multivariate analysis, a new nomogram model was constructed using the RStudio 3.4 software. Results:(1) Celiac lymph node metastasis in TE-SCC: celiac lymph node metastasis was found in 89 of the 443 patients, with a celiac lymph node metastasis rate of 20.09%(89/443). (2) Analysis of influencing factors for celiac lymph node metastasis in TE-SCC. Results of univariate analysis showed that tumor location, tumor length, tumor differentiation degree, pathological T staging, nerve invasion, vessel invasion, and thoracic lymph node metastasis were related factors for celiac lymph node metastasis in TE-SCC ( χ2=12.177, Z=-2.754, -4.218, -4.254, χ2=3.908, 33.025, 30.387, P<0.05). Results of multivariate analysis showed that tumor location, vessel invasion, and thoracic lymph node metastasis were independent influencing factors for celiac lymph node metastasis in TE-SCC ( odds ratio=2.165, 3.442, 2.876, 95% confidence interval: 1.380-3.396, 1.787-6.633, 1.631-5.071, P<0.05). (3) Construction of nomogram prediction model of celiac lymph node metastasis in TE-SCC: based on the factors screened by multivariate analysis, including tumor location, vessel invasion, and thoracic lymph node metastasis, the nomogram prediction model of celiac lymph node metastasis in TE-SCC was established, with the concordance index of 0.846. The calibration curve showed a high consistency between the celiac lymph node metastasis probability estimated by the prediction model and the actual rate of celiac lymph node metastasis. The decision curve showed that the nomogram prediction model of celiac lymph node metastasis in TE-SCC had a good prediction value when the probability threshold was 0.001-0.819.(4) Construction of decision-making tree model of celiac lymph node metastasis in TE-SCC and risk subgroup analysis of celiac lymph node metastasis probability: patients were stratified into six risk subgroups using the decision-making tree model based on the celiac lymph node metastasis probability. The group A included patients with no vessel invasion+negative thoracic lymph node, group B included patients with no vessel invasion+the number of positive thoracic lymph nodes of 1-3, group C included patients with no vessel invasion+the number of positive thoracic lymph nodes of ≥4, group D included patients with vessel invasion+the number of positive thoracic lymph nodes of 0-2+upper or middle thoracic esophageal carcinoma, group E included patients with vessel invasion+the number of positive thoracic lymph nodes of 0-2+lower thoracic esophageal carcinoma, group F included patients with vessel invasion+the number of positive thoracic lymph nodes of ≥3. The group A was low-risk group with the celiac lymph node metastasis probability of 11%, group B and D were intermediate low-risk groups with the celiac lymph node metastasis probability of 27% and 21%, group C and E were the intermediate high-risk groups with the celiac lymph node metastasis probability of 56% and 55%, and group F was high-risk group with the celiac lymph node metastasis probability of 80%. Conclusions:The tumor location, vessel invasion, and thoracic lymph node metastasis are independent influencing factors for celiac lymph node metastasis in TE-SCC. Vessel invasion has the dominant influence on celiac lymph node metastasis, followed by the number of positive thoracic lymph nodes, and then the tumor location. Patients can be stratified into six risk subgroups based on the nomogram prediction model and decision-making tree model of celiac lymph node metastasis in TE-SCC.

TSCI have dyskinesia and sensory disturbance that can cause various life-threaten complications. The patients with traumatic spinal cord injury (TSCI), seriously affecting the quality of life of patients. Based on the epidemiology of TSCI and domestic and foreign literatures as well as expert investigations, this expert consensus reviews the definition, injury classification, rehabilitation assessment, rehabilitation strategies and rehabilitation measures of TSCI so as to provide early standardized rehabilitation treatment methods for TSCI.

Objective To investigate the difference between histopathological changes of brain white matter in low-density lipoprotein receptor (LDLR) homozygous mutation rats with hypercholesterolemia and wild-type rats.Methods Thirty LDLR-/-rats and 28 wild-type rats were selected.Plasma cholesterol levels were measured by enzyme-linked immunosorbent assay at 15,18 and 26 weeks old respectively.The axonal structure of the corpus callosum area was observed by transmission electron microscopy.The myelin basic protein (MBP) of the corpus callosum area was quantitatively analyzed by Western blotting.In addition,at 26 weeks old,the myelin sheaths were stained by fast blue staining.The expression level of MBP in white matter was further detected by immunofluorescence staining,and the morphological changes of glial cells were observed.Results Compared with the wild-type rats,the plasma cholesterol concentration in LDLR-/-rats increased significantly,and it could be as high as 3.3 times at 26 weeks.The results of electron microscopy showed that the LDLR-/-rats had axonal injury at 15 weeks and aggravated gradually over time.At 26 weeks,Western blot analysis of the LDLR-/-rats showed that the MBP expression level of the corpus callosum area decreased significantly.Fast blue staining showed loosening of nerve fibers,diffuse vacuole formation,and myelinated nerve fiber loss in the corpus callosum area.In addition,it was also found that the number of oligodendrocytes in LDLR-/-rats was significantly reduced,and large numbers of astrocytes and microglia were activated.Conclusions LDLR-/-rats will have spontaneous hypercholesterolemia.Axonal injury,demyelination,decreased oligodendrocytes,as well as the abnormal activation of astrocytes and microglia are present in the early adult brain white matter area.

Objective To investigate the predictive value of serum adiponectin (APN) for post-stroke depression (PSD) in patients with ischemic stroke. Methods Patients with acute ischemic stroke admitted to the Departments of Neurology, the Affiliated Brain Hospital of Guangzhou Medical University, and Dongguan Changping Hospital were enrolled prospectively from September 2016 to January 2018. Serum APN levels were measured by radioimmunoassay the next day after admission. PSD was diagnosed by using the Diagnostic and Statistical Manual of Mental Disorders (4th Edition) at one month after discharge. The baseline clinical features in the PSD group and the non-PSD group were compared. Multivariable logistic regression analysis was used to determine the risk factors for PSD. The receiver operating characteristic (ROC) curve was used to analyze the predictive value of APN for PSD. Results A total of 220 patients were enrolled, including 57 (25.9%) in the PSD group and 163 (74.1%) in the non-PSD group. The proportion of patients with diabetes (P=0.020) and living alone (P=0.012), as well as age (P=0.17), homocysteine (P=0.009), C-reactive protein ( P=0.001), and National Institutes of Health Stroke Scale score ( P=0.007) in the PSD group were significantly higher than those in the non-PSD group, while the APN level was significantly lower than that in the non-PSD group (P=0.003). Multivariate logistic regression analysis showed that the 4th quartile of serum APN level was used as a reference, the 1st quartile of APN level was an independent risk factor for PSD (odds ratio, 4 .202, 95% confidence interval 1.401-12.067; P=0.013). ROC curve analysis showed that the area under the curve of serum APN level for predicting PSD was 0.642 (95% confidence interval 0.564-0.721; P=0.001). The optimal cutoff value was 6.4 mg/L, and its sensitivity of predicting PSD was 63.2%, the specificity was 63.8%, the positive predictive value was 83.3%, and the negative predictive value was 57 .7%. Conclusion Serum APN has a certain predictive value for PSD in patients with ischemic stroke.

Objective To observe bone formation of compound of autologous periosteum cells plus calcium alginate gelatin in animal body and investigate the best mixing ratio of the two components.Methods A total of 96 healthy adult New Zealand rabbits were randomly divided into six groups (Groups A,B,C,D,E and control group).The excised bilateral periostea from rabbits in each experimental group were all made into 0.4 ml cell suspensions.The compounds were prepared by blending 0.4 ml calcium alginate gelatin with 1,1/4,1/16,1/64 and 1/256 of the periosteum cell suspension respectively,and then were applied to autologous right radial defect area.Gross observation,X-ray examination and histological study were carried out at 2,4,8,12 weeks postoperatively.Levels of serum alkaline phosphatase and serum calcium were determined as well.Results The compounds containing periosteum cell suspension with cell counts of 1/4 or 1/16 and calcium alginate gelatin reached the best osteogenesis.Conclusion Compound of autologous periosteum cell suspension-calcium alginate gelatin induces obvious bone formation and is worthy of clinical application,for it has advantages of satisfactory bone defect repair and easy operation.

Cloning of flanking sequences of double-copy gene is a challenge in molecular biology. We developed a method to solve this problem by combining an optimized inverse PCR (iPCR) with TAIL-PCR. First, Southern blotting analysis was used to determine a proper restriction enzyme that could obtain proper-length restriction fragments that contained the target gene. Then optimized iPCR was performed to amplify the restriction fragments that contained the separated copies of the gene. Based on the obtained sequences, TAIL-PCR was performed to amplify further flanking regions of the gene. With this method, we obtained all of the EcoR I restriction fragments (2.2-5.1 kb) and Hind III restriction fragments (8.5-11.7 kb) of mitochondrial atpA gene in cytoplasmic male sterile (CMS) line and maintainer line of Upland cotton. The results showed that this method was an efficient approach to clone flanking sequences of double-copy gene.
Chromosome Walking ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Genes, Mitochondrial ; Genes, Plant ; genetics ; Gossypium ; genetics ; Plant Proteins ; genetics ; metabolism ; Polymerase Chain Reaction ; methods ; Terminal Repeat Sequences

BACKGROUND: Human adipose tissue-derived stromal cells (ADSCs) possess exert proliferation and multi-directional ability. As a novel stem cell, it has been widely utilized in tissue engineering and plays an important role in biological and potential therapeutic applications.OBJECTIVE: To review the research progress, applications and existing problems of human ADSCs in tissue engineering and cell therapy by retrieving relevant publications. METHODS: PubMed and CNKI databases were undertaken to identify the relevant articles published from January 1960 to January 2009 with the key words of "human adipose tissue-derived mesenchymal cells, isolation, differentiation, immune phenotype, application" both in English and Chinese. The articles relate to biological characteristics and applications of ADSCs were selected. In the same field, the documents published earlier or in the authoritative journals were preferred, and the repetitive studies were excluded. RESULTS AND CONCLUSION: A total of 81 documents were searched by computer, and 57 references were included in the final analysis. The results demonstrated that, human ADSCs share many characteristics, including the high proliferative potential and exhibiting the exert ability to undergo multilineage differentiation under appropriate conditions. Human ADSCs can not only utilize for tissue repairing, but also for cell immune modulation and gene therapy. However, there are still problems in its application. With the development of research on human ADSCs, their biological characteristics will be revealed, and their application in tissue repairing, cell therapy, transplantation, as well as gene therapy must be hold a great promise.