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.

Objective Tostudy the influence of pipe structures on the mixing uniformity of airborne effluents from nuclear power plant chimneys. Methods We used the computational fluid dynamics (CFD) method to simulate the velocity distribution and gas mixing in long straight pipes (I type) with square section and circular section, 90° single-bend pipes (L type) with square section and circular section, and 90° double-bend pipes (S type and U type) with square section and circular section. Results For the long straight pipe, due to the lack of flow disturbance caused by structural changes, the mixing effect was not good; when the pipe section was circular, it might take mixing distance 20 times the hydraulic diameter to achieve the uniformity index required by the relevant standard; for the square pipe, the distance might be longer. In the single bend pipe with square section, the velocity uniformity was improved more greatly after the bend, and the tracer gas met the mixing uniformity at a shorter distance (11 times the hydraulic diameter), as compared with the single bend pipe with circular section. For the S-type double-bend pipe, the tracer gas appeared uniformly mixed after a distance 6 times the hydraulic diameter in the square pipe, and 7 times the hydraulic diameter in the circular pipe. For the U-type double-bend pipe, the gas in the square pipe also achieved uniform mixing ata shorter distance downstream, and the airflow showed greater disturbance when passing through the bend. Conclusion The CFD method can make an accurate prediction for the change patterns of gas mixing uniformity in pipes with different structures, and can partially replace physical experiments to study the factors affecting the mixing uniformity of airborne effluents from the chimney of nuclear power plants.

OBJECTIVE: To analyze the clinical and genetic features of three patient diagnosed with Kleefstra syndrome. METHODS: Whole exome sequencing (WES) was carried out for the probands and their parents. Suspected variants were validated by Sanger sequencing. Copy number variations (CNV) were detected by CNV-seq and validated by real-time PCR. RESULTS: Proband 1 was found to carry a de novo heterogeneous variant (c.823+1G>T) of the EHMT1 gene, which may affect its expression. Based on the guidelines of the American College of Medical Genetics and Genomics, the variant was predicted to be pathogenic (PVS1+PS2+PM2). Proband 2 was found to carry a de novo missense variant c.439C>G (p.L147V) of the EHMT1 gene, which was predicted to be likely pathogenic (PS2+PM1+PM2+PP3). Proband 3 was found to carry a heterozygous 520 kb deletion at 9q34.3 by CNV-seq. The deletion has encompassed the whole of the EHMT1 gene. Real-time PCR has detected no CNV of this region in her parents. CONCLUSION Variants of the EHMT1 gene probably underlay the disease in these patients. Genetic testing has provided a basis for their clinical diagnosis.
Chromosome Deletion ; Chromosomes, Human, Pair 9 ; Craniofacial Abnormalities ; DNA Copy Number Variations ; Female ; Genetic Testing ; Heart Defects, Congenital ; Humans ; Intellectual Disability/genetics* ; Mutation

COVID-19/virology* ; China/epidemiology* ; Disease Outbreaks ; Genetic Linkage ; Humans ; Phylogeny ; Polymorphism, Single Nucleotide ; SARS-CoV-2/isolation & purification*

An unexpected observation among the COVID-19 pandemic is that smokers constituted only 1.4%-18.5% of hospitalized adults, calling for an urgent investigation to determine the role of smoking in SARS-CoV-2 infection. Here, we show that cigarette smoke extract (CSE) and carcinogen benzo(a)pyrene (BaP) increase ACE2 mRNA but trigger ACE2 protein catabolism. BaP induces an aryl hydrocarbon receptor (AhR)-dependent upregulation of the ubiquitin E3 ligase Skp2 for ACE2 ubiquitination. ACE2 in lung tissues of non-smokers is higher than in smokers, consistent with the findings that tobacco carcinogens downregulate ACE2 in mice. Tobacco carcinogens inhibit SARS-CoV-2 spike protein pseudovirions infection of the cells. Given that tobacco smoke accounts for 8 million deaths including 2.1 million cancer deaths annually and Skp2 is an oncoprotein, tobacco use should not be recommended and cessation plan should be prepared for smokers in COVID-19 pandemic.
Adult ; Animals ; COVID-19 ; Epithelial Cells ; Humans ; Lung ; Mice ; Pandemics ; Peptidyl-Dipeptidase A ; SARS-CoV-2 ; Spike Glycoprotein, Coronavirus ; Ubiquitin-Protein Ligases/genetics*

Objective:The risk points of highly pathogenic virus culture in BSL-3 laboratory in basket bioreactor were evaluated, and the safety of virus culture in BSL-3 laboratory was judged by monitoring virus concentration in three stages of virus inoculation, culture and harvest.Methods:Smear samples were taken from the tank and joints of the basket biorereactor at three stages of virus inoculation, culture and harvest with sterilized swabs. At the same time, in the biological safety cabinet, air samples were taken for virus inoculation and harvesting operations using AirPortMD8 air sampler, and in the three stages after operation, the personal protective equipment of researchers was to apply sampling, with three ways to monitoring.Results:During the three stages of virus inoculation, culture and receipt, the virus test result of samples taken in the tank and the junction of the basket bioreactor were all negative, the virus test result of air samples taken during the operation stage of inoculation and receipt were all negative, and the virus test result of samples taken in the personal protective equipment of the experimenter were all negative.Conclusions:The basket bioreactor method for virus culture in BSL-3 laboratory is safe and reliable, and the test result show that there is no virus leakage during the whole operation process, thus ensuring the biosafety of experimenter and laboratory environment.

Objective To investigate the effect of high-flow nasal cannula oxygen therapy (HFNC) on the clinical efficacy and diaphragm function of patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). Methods The patients with mild to moderate AECOPD (clinical classification Ⅰ-Ⅱ) admitted to Huxi Hospital Affiliated to Jining Medical College from January to October in 2018 were enrolled. The patients were divided into HFNC treatment group and routine oxygen therapy control group (each n = 37) by randomly number table method. The two groups were given bronchiectasis drugs, corticosteroids, expectorant, anti-infection treatment, at the same time, the HFNC treatment group was given HFNC with the initial flow rate of 40 L/min. The routine oxygen therapy control group was given low flow oxygen, and the initial flow rate was 3 L/min. General data such as gender, age, clinical grade, acute physiology and chronic health evaluationⅡ (APACHEⅡ) score were recorded. Bedside ultrasound was used to measure the diaphragmatic excursions during quiet breathing (DEq), diaphragmatic excursions during deep breathing (DEd), and diaphragmatic shallow fast breathing index (D-RSBI) before and 2, 24 and 48 hours after treatment in both groups and compared, meanwhile, arterial blood gas analysis was performed, and arterial partial pressure of oxygen (PaO2) and arterial partial pressure of carbon dioxide (PaCO2) were recorded. Results Two patients in the HFNC treatment group withdrew from the study because they could not tolerate HFNC, while other patients were enrolled in the analysis. There was no statistically significant difference in gender, age, proportion of AECOPDⅡ grade or APACHEⅡscore between the two groups, indicating that the general data of the two groups were comparable and balanced. There was no statistically significant difference in DEq, DEd, D-RSBI, PaO2 or PaCO2 before treatment between the two groups. After treatment, DEp in both groups was decreased gradually with time, it was decreased earlier in the HFNC treatment group, and it showed significant difference as compared with that before treatment at 2 hours after treatment (mm: 18.3±3.1 vs. 20.1±4.2, P < 0.01), and it was significantly lower than that in the routine oxygen therapy control group (mm: 18.3±3.1 vs. 20.3±3.7, P < 0.05); DEd was gradually increased in both groups, it was significantly increased in the HFNC treatment group, and it was significantly higher than that in the routine oxygen therapy control group at 24 hours and 48 hours after treatment (mm: 55.2±7.6 vs. 50.8±9.2 at 24 hours, 59.4±7.7 vs. 53.6±9.1 at 48 hours, both P < 0.05); D-RSBI was decreased gradually in both groups, it was decreased earlier and more significant in the HFNC treatment group, and it was significantly lower than that in routine oxygen therapy control group at 24 hours and 48 hours after treatment (times·min-1·mm-1: 0.41±0.13 vs. 0.51±0.20 at 24 hours, 0.31±0.12 vs. 0.43±0.17 at 48 hours, both P < 0.05). After treatment, there was no statistically significant difference in PaO2 or PaCO2 between the two groups. Conclusion HFNC can effectively relieve diaphragm fatigue in patients with mild to moderate AECOPD, but it had no effect on carbon dioxide retention.

OBJECTIVE： To systematically evaluate the efficacy and safety of TCM compound preparation for tonifying kidney and activating blood circulation， and to provide evidence-based reference for rational drug use in the clinic. METHODS： By retrieving Cochrane library， PubMed， Embase， CBM， CNKI， VIP and Wanfang database， randomized controlled trials （RCTs） about TCM compound preparation for tonifying kidney and activating blood circulation （trial group） versus calcium or non-calcium agents （control group） in the treatment of postmenopausal osteoporosis were included. After literature screening， data extraction and quality evaluation with bias risk evaluation tool and Jadad scale of Cochrane system evaluator manual 5.1.0， Meta-analysis was conducted by using Stata 12.0 software， and trial sequential analysis （TSA） was conducted by using TSA 0.9 software. RESULTS： Totally 18 RCTs were included， involving 1 408 patients. The results of Meta-analysis showed that total response rate [RR＝1.35，95％CI（1.17，1.54），P＜0.000 1] and bone density[SMD＝0.24，95％CI（0.16，0.32），P＜0.000 1] of trial group were significantly higher than those of control group； blood calcium [SMD＝－0.05，95％CI（－0.09，0.00）， P＝0.033] of trial group was significantly lower than that of control group. There was no statistical significance in the levels of urine creatinine [SMD＝－1.60，95％CI（－5.94，2.74），P＝0.470]， urinary calcium/urine creatinine ratio [SMD＝－0.05，95％CI（－0.14，0.04），P＝0.295]， urinary hydroxyproline/urine creatinine ratio [SMD＝－0.16，95％CI（－1.04，0.72），P＝0.726]， ALT [SMD＝0.51，95％CI（－3.26，4.28），P＝0.790]， AST [SMD＝0.23，95％CI（－5.22，4.77），P＝0.929]， serum alkaline phosphatase [SMD＝－0.22，95％CI（－0.68，0.25），P＝0.361]， serum phosphate [SMD＝－0.02，95％CI（－0.11，0.07），P＝0.639]， urea nitrogen [SMD＝－0.19，95％CI（－0.70，0.31），P＝0.453]， estradiol [SMD＝0.62，95％CI（－0.28，1.52），P＝0.177]， IL-6 [SMD＝－1.78，95％CI（－4.86，1.30），P＝0.258] or VAS [SMD＝0.55，95％CI（－1.03，2.13），P＝0.496] between 2 groups. No server ADR was found in 2 groups. TSA showed that there were extract evidences for total response rate of TCM compound preparation in the treatment postmenopausal osteoporosis. CONCLUSIONS： TCM compound preparation for tonifying kidney and activating blood circulation shows significant therapeutic efficacy for postmenopausal osteoporosis， and can improve serum calcium and bone density with good safety.

We expressed multi-epitope chimeric protein of CARDS toxin protein of Mycoplasma pneumonia (Mp) in prokaryotic cells,and purified and investigated its immunoreactivity.A recombinant multi-epitope chimeric gene including ten critical epitopes was connected by linker and cloned into prokaryotic expression vector pET-28a(+),and transformed into E.coli BL21(DE3) cells for expression under induction of IPTG.The antigenicity of expressed recombinant protein was identified with 6 × His monoclonal antibody and human positive serum by Western blot.The recombinant expression vector pET-CARDS was constructed and the about 30 kDa recombinant chimeric protein expressed in BL21(DE3) successfully.Western blot analysis showed that it can react respectively with 6 × His monoclonal antibodies and human positive serum.This study showed that the chimeric CARDS protein has an obvious immunoreactivity and a potential to be a new antigen for the diagnosis of Mp infection.

Objective To observe the therapeutic effect ofYiqi-Fumai injection andShenshao capsule in the treatment of angina pectoris and the influence on vascular endothelial cell function and serum MPO. Methods 90 cases of angina pectoris of coronary heart disease were randomly divided into a treatment group and a control group. 50 cases in the treatment group were treated withYiqi-Fumai injection and Shenshao capsule; 40 cases in the control group were treated with Plavix and metoprolol; clinical symptoms were observed before and after treatment, the indexes of endothelial cell function and electrocardiogram blood serum nitric oxide (NO), endothelin (ET), NOS and serum MPO, SOD, GSH changes were compared.Results The content of nitric oxide (NO), 6-keto-PGFLa and NOS of the treatment group were increased than the control group, while ET was reduced. In the treatment group, NO was (78.09 ± 88.71)μmol/L 6-keto-PGFL was 32.04 ± 25.82 pg/ml and NOS was 27.88 ± 9.08 U/MI before the treatment,; after the treatment, the NO was 127.11 ± 103.60μmol/L, 6-keto-PGFLa was 50.03 ± 60.87 pg/ml and NOS was 35.12 ± 22.18 U/MI. In the control group, NO was 96.98 ± 82.45μmol/L, 6-keto-PGFLa was 40.31 ± 51.92 pg/ml and NOS was 28.12 ± 5.11 U/MI before the treatment; After the treatment, NO was 95.62 ± 6.31μmol/L, 6-keto-PGFLa was 29.09 ± 42.01 pg/ml and NOS was 30.11 ± 7.51 U/MI. In the treatment group, ET was 58.81 ± 33.18 pg/ml before the treatment and 42.47 ± 40.5 pg/ml. after the treatment. In the control group, ET was 56.81 ± 32.76 pg/ml before the treatment and 60.67 ± 12.81 pg/ml after the treatment. In terms of resistance to oxidative stress and MPO, the SOD ,GSH and MPO improvement rate of the treatment group was significantly better than the control group (P<0.05). In the treatment group, SOD was 48.36 ± 17.38 U/MI, GSH was 126.68 ± 19.93 g/L and MPO was 57.33 ± 8.34 mmol/L before the treatment, after the treatment, SOD was 62.45 ± 15.76 U/MI, GSH was 135.98 ± 28.99 g/L and MPO was 17.11 ± 3.60 mmol/L. In the control group, SOD was 48.44 ± 19.37 U/MI, GSH was 29.44 ± 21.66 g/L and MPO was 56.35 ± 14.44 mmol/L before the treatment, after the treatment, SOD was 57.35 ± 14.44 U/MI, GSH was 28.56 ± 24.06 g/L and MPO was 48.62 ± 6.31 mmol/L.ConclusionYiqi-Fumai injection combined with Shenshao capsule has curative effect on angina pectoris of coronary heart disease, being worth of clinical use.