Development of biosensors highly responsive to N-acetylneuraminic acid in Bacillus subtilis / 生物工程学报

Bacillus subtilis is recognized as a generally-regarded-as-safe strain, and has been widely used in the biosynthesis of high value-added products, including N-acetylneuraminic acid (NeuAc) which is widely used as a nutraceutical and a pharmaceutical intermediate. Biosensors responding to target products are widely used in dynamic regulation and high-throughput screening in metabolic engineering to improve the efficiency of biosynthesis. However, B. subtilis lacks biosensors that can efficiently respond to NeuAc. This study first tested and optimized the transport capacity of NeuAc transporters, and obtained a series of strains with different transport capacities for testing NeuAc-responsive biosensors. Subsequently, the binding site sequence of Bbr_NanR responding to NeuAc was inserted into different sites of the constitutive promoter of B. subtilis, and active hybrid promoters were obtained. Next, by introducing and optimizing the expression of Bbr_NanR in B. subtilis with NeuAc transport capacity, we obtained an NeuAc-responsive biosensor with wide dynamic range and higher activation fold. Among them, P535-N2 can sensitively respond to changes in intracellular NeuAc concentration, with the largest dynamic range (180-20 245) AU/OD. P566-N2 shows a 122-fold of activation, which is 2 times of the reported NeuAc-responsive biosensor in B. subtilis. The NeuAc-responsive biosensor developed in this study can be used to screen enzyme mutants and B. subtilis strains with high NeuAc production efficiency, providing an efficient and sensitive analysis and regulation tool for biosynthesis of NeuAc in B. subtilis.

Basic researches over spinal cord injury: problems and countermeasures / 中华创伤杂志

Spinal cord injury leads to extremely high mortality and disability rates and its treatment has always been a global challenge. The survival of patients with spinal cord injury is partially lengthened with the development of medical treatments, but the clinical outcome is still not satisfactory. Some important progress has been made in the basic researches over spinal cord injury, such as analysis of repair mechanism of spinal cord injury and development of cellular therapies and biological scaffolds of spinal cord injury in China. However, some basic researches show insufficient understanding of the microenvironment and animal model of spinal cord injury and lack support from clinical problems, leading to too simplistic or contradictory conclusions. There is an urgent need to reexamine the research methods and carry out basic and clinical translational researches. Therefore, the authors discuss the key problems and difficulties in basic researches over spinal cord injury and propose improvement suggestions, aiming to provide a reference for conducting basic researches correctly and accelerating clinical innovational transformation.

Features of severe acute respiratory syndrome coronavirus 2 co-infected with other common respiratory pathogens in Shanghai City, 2020-2021 / 中华传染病杂志

Objective:To analyze the features of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) co-infected with other common respiratory pathogens among coronavirus disease 2019 (COVID-19) patients in Shanghai City, and to provide a reference for scientific prevention and control of COVID-19 and other respiratory infectious diseases.Methods:Descriptive epidemiological approaches were used to analyze the data of COVID-19 reported cases in Shanghai City from January 2020 to February 2021 in the information system of Chinese Disease Prevention and Control. Clinical data of the participants were collected, and their SARS-CoV-2 nucleic acid-positive respiratory specimens were collected at the time of illness onset or admission. Multiplex reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect the 22 respiratory pathogens. Independent-samples t test was used for statistical analysis. Results:Of the 272 patients with COVID-19, 15(5.5%) had co-infection of SARS-CoV-2 with other respiratory pathogens, all of which were double infection. There were three cases infected with enterovirus/rhinovirus, two of each with adenovirus, human metapneumovirus and coronavirus NL63/HKU1, and one of each with coronavirus 229E, influenza A virus H1N1, parainfluenza virus 1 and respiratory syncytial virus B. Two cases infected with Mycoplasma pneumoniae. Among the 272 COVID-19 patients, 212(77.9%) had fever, 117(43.0%) had cough, 46(16.9%) had fatigue, and 35(12.9%) had sore throat. The white blood cell count of co-infection cases was higher than that of non-co-infection cases ((6.8±1.7)×10 9/L vs (5.3±1.6)×10 9/L), and the difference was statistically significant ( t=3.09, P=0.008). Conclusions:There is a certain proportion of co-infection of SARS-CoV-2 with other respiratory pathogens among the COVID-19 cases in Shanghai City, mainly viral pathogens, especially enterovirus/rhinovirus. A rational combination of drugs was recommended to improve the cure rate. Surveillance of acute respiratory infection should be further strengthened as well.

Effects of preoperative epidemiological factors on the surgical outcomes of benign prostatic hyperplasia / 现代泌尿外科杂志

【Objective】 To investigate the preoperative epidemiological factors affecting the surgical outcomes of transurethral resection of the prostate (TURP) in patients with benign prostatic hyperplasia (BPH) and to evaluate the clinical predictive value. 【Methods】 Patients undergoing TURP at our hospital during Dec.2018 and Sep.2021 whose pathological examination suggestive of BPH were involved. Before operation, the clinical data, International Prostate Symptom Score (IPSS), quality of life (QoL) score, overactive bladder (OAB) score, prostate-specific antigen (PSA), and urological ultrasound results were collected. After operation, all patients were followed up with telephone call or face-to-face interview, and the IPSS, QoL score and OAB score were examined. Patients were divided into effective and ineffective groups according to the efficacy rubrics. The epidemiological factors affecting the efficacy of TURP were identified with univariate analysis, the independent influencing factors were screened with binary logistic regression, and the diagnostic value of each independent influencing factor was evaluated using receiver operating characteristic (ROC) curve. 【Results】 OAB score (OR=0.749, 95%CI: 0.627-0.895, P=0.001), IPSS-V/S (OR=4.919, 95%CI: 1.617-14.963, P=0.005), history of urinary retention (OR=7.513, 95%CI: 2.289-24.656, P=0.001), and history of urinary incontinence (OR=2.656, 95%CI: 1.015-6.950, P=0.047) were independent influencing factors for poor postoperative outcomes. ROC curve revealed that the area under the ROC curve (AUC) showed that AUC of OAB score, IPSS-V/S, history of urinary retention, and history of urinary incontinence were 0.784 (95%CI: 0.718-0.841), 0.686 (95%CI: 0.614-0.751), 0.713 (95%CI: 0.643-0.777), and 0.723 (95%CI: 0.654-0.786), respectively. ROC curve of the regression model showed that the AUC was 0.888 (95%CI: 0.834-0.930), and the sensitivity and specificity were 93.53% and 67.35%, respectively. 【Conclusion】 Preoperative OAB score, IPSS-V/S, history of urinary retention, and history of urinary incontinence were independent epidemiological factors of poor outcomes after TURP in BPH patients. The diagnostic efficacy ranked from the highest to the lowest as regression model >OAB score>history of urinary incontinence >history of urinary retention>IPSS-V/S.

Effect of online and offline collaborative and integrated mode on voluntary blood donation: Internet + blood donation souvenir / 中国输血杂志

【Objective】 To explore the effect of online and offline collaborative and integrated mode (Internet + blood donation souvenirs), and develop diversified scientific ways of caring for voluntary blood donors in combination with the potential expectation of the target population, ao as to recruit and retain more blood donors. 【Methods】 A total of 4 000 blood donors from Yichang Central Blood Station from January 1 to June 30, 2022 were selected, and they could collect souvenirs from online and offline. The intelligent platform V9.5 of modern blood station management information system of Qiao Technology was used to classify the data of online and offline souvenir distribution, and digital intelligent comparison and analysis on age, occupation, education, type (first-time blood donors or repeat blood donors), region (main urban districts of Yichang and other counties and suburban districts) and gender were carried out. 【Results】 A total of 1 400 blood donors chose online souvenirs, who were mainly under 25 years old, students, female, college education or above, urban areas and regular donors; 2 600 chose offline souvenirs, who were mainly over 25 years old, male, junior college degree or below, occupations other than students (medical workers, educators, etc.), counties and suburban districts, and first-time blood donors. The collaborative and integrated mode of the two distribution methods were complementary and had a good incentive effect on blood donor recruitment. 【Conclusion】 The online and offline collaborative and integrated mode demonstrates a good effect and meets the needs of different population. Digital intelligence system is helpful to develop diversified and scientific ways of caring for voluntary blood donors, recruit and retain more blood donors, achieve high-quality development of blood collection and supply, therefore guarantee the increasing demand for clinical blood use.

Relationship between job stress/sleep quality and job satisfaction among programmers in a Shanghai Internet enterprise / 上海预防医学

ObjectiveTo study the relationship between occupational stress/sleep quality and job satisfaction of internet enterprise programmers， to explore the mediating effect of sleep quality between occupational stress and job satisfaction， and to provide a theoretical basis for taking effective measures to improve job satisfaction. MethodsA total of 271 programmers from an Internet company in Shanghai were investigated by job content questionnaire， Pittsburgh Sleep Index Scale and the third part of the McLean occupational stress questionnaire. The results were statistically analyzed ResultsThe positive rate of occupational stress of programmers in this enterprise was 66.7%， 34.3% of them had average sleep quality， 28.0% had poor sleep quality， and the score of job satisfaction was 47.70±6.78. Correlation analysis showed that the higher the occupational stress， the worse the sleep quality and the lower the job satisfaction. Effects of occupational stress and sleep quality on job satisfaction were both statistically significant. The total effect of occupational stress on job satisfaction was -0.35， and the mediating effect of sleep quality between occupational stress and job satisfaction was -0.04， accounting for 12.0% of the total effect. ConclusionSleep quality of internet enterprise programmers plays a partial mediating role between occupational stress and job satisfaction， and a series of measures such as improving sleep can be taken to improve programmer’s job satisfaction.

Difference analysis of 18F-FMISO PET/CT hypoxia imaging in response to heavy ion radiotherapy in patients with non-small cell lung cancer / 中华核医学与分子影像杂志

Objective:To explore the clinical value of 18F-fluoromisonidazole (FMISO) PET/CT hypoxia imaging in early response to heavy ion radiotherapy in patients with non-small cell lung cancer(NSCLC). Methods:From April 2018 to January 2021, the 18F-FMISO PET/CT images of 23 NSCLC patients (19 males, 4 females; age (64.9±10.3) years) who received heavy ion radiotherapy in Shanghai Proton and Heavy Ion Center were retrospectively analyzed. The evaluation parameters included tumor volume (TV), tumor to background ratio (TBR) before and after radiotherapy. Patients were divided into hypoxia group and non-hypoxia group with the baseline TBR value≥1.4 as hypoxia threshold. Wilcoxon signed rank test was used to compare the differences of TV and TBR before and after radiotherapy in 2 groups. Results:Of 23 NSCLC patients, 17 were hypoxia and 6 were non-hypoxia. Compared with the baseline, TV after the radiotherapy (59.44(22.86, 99.43) and 33.78(8.68, 54.44) cm 3; z=-3.05, P=0.002) and TBR after the radiotherapy (2.25(2.09, 2.82) and 1.42(1.24, 1.67); z=-3.39, P=0.001) of the hypoxia group were significantly lower, while TV (16.19(6.74, 36.52) and 8.59(4.38, 25.47) cm 3; z=-1.57, P=0.120) and TBR (1.19(1.05, 1.27) and 1.10 (0.97, 1.14); z=-1.89, P=0.060) of the non-hypoxia group decreased with no significant differences. Conclusions:Hypoxic NSCLC tumors are sensitive to heavy ion radiation. Compared with non-hypoxic tumors, hypoxic tumors respond more quickly, and a significant reduction in TV can be observed early after radiotherapy. Heavy ion radiation can significantly improve tumor hypoxia.

Prevalence of respiratory and allergic diseases in children in Xuhui District， Shanghai in 2013 and 2020 / 上海预防医学

ObjectiveTo compare the incidence of respiratory and allergic diseases in children in Xuhui District， Shanghai in 2013 and 2020， and to determine the influencing factors. MethodsAnnual average levels of air pollutants including PM2.5， PM10， O3， SO2， and NO2 were collected and described in Shanghai from 2013 to 2020. A cross-sectional survey was conducted by using a questionnaire among grade 3 to 5 students in a school in Xuhui District， Shanghai， in September 2013 and 2020， respectively. The questionnaire collected variables including living environment， daily habits， family history of respiratory and allergic diseases， and incidence of these diseases in children. Chi-square test was used to determine the difference across respiratory and allergic diseases. Logistic regression was conducted to determine the influencing factors. ResultsA total of 1 398 valid questionnaires were collected （705 in 2013 and 693 in 2020）. Compared with 2013， annual average concentrations of PM2.5， PM10， O3， SO2， and NO2 in 2020 significantly decreased. The prevalence of bronchial asthma， bronchitis， persistent cough and persistent expectoration in 2013 were significantly higher than those in 2020 （P<0.05） in Xuhui District. Multivariate analysis showed that severe air pollution， boys， parents with asthma or allergy， parents with higher educational levels， and more allergens in household were the risk factors associated with the incidence of bronchial asthma， bronchitis， allergic rhinitis and atopic eczema （P<0.05）. Parents with allergy history， high smoking frequency of family member， and more allergens in household were the risk factors associated with the incidence of persistent cough and persistent expectoration （P<0.05）. ConclusionTo 2013，2020 air pollution in Shanghai has been mitigated and prevalence of bronchial asthma and bronchitis of children has decreased. Childhood respiratory and allergic diseases are associated with indoor and outdoor environment， family medical history， and family daily habits.

To explore the quality markers of Huangqin Qingre Chubi capsule in the treatment of rheumatoid arthritis based on network pharmacology combined with pharmacokinetics and target verification / 药学学报

As an effective prescription for the treatment of rheumatoid arthritis (RA), Huangqin Qingre Chubi capsule (HQC) is still blank in quality control. This study aims to explore quality markers (Q-markers) for HQC in the treatment of RA by integrating network pharmacology and pharmacokinetics. By constructing the visualization network of "pharmacodynamic ingredient-target-pathway", the potential Q-Marker of HQC treatment for RA was preliminatively predicted. A rat model of rheumatic heat obstruction syndrome collagene-induced arthritis (CIA) was established to elucidate the dynamic quantification law of pharmacodynamic components of HQC in the disease state of rats. To establish the inflammatory model of RA synovial fibroblasts (MH7A) induced by tumor necrosis factor-α (TNF-α) in vitro. The effects of active ingredients on protein expression of sphingosin kinase-1 (Sphk1) and p-SphK1 were detected. The network pharmacological results showed that baicalin, geniposide, luteolin, coixol and amygdalin were the important active components of HQC treatment for RA. Quantitative analysis results further verified the measurability of these five components. The expression of Sphk1 and p-SphK1 was significantly inhibited by geniposide and baicalin by Western blotting. The above studies determined that the above 5 components could be used as Q-markers in the treatment of RA by HQC. This experiment was approved by the Experimental Animal Ethics Committee of Anhui University of Chinese Medicine (approval number: AHUCM-rats-2021049). All procedures were conducted in strict accordance with the principles of animal use and care.

Thoracoscopic combined subsegmentectomy for 76 patients: A retrospective study in a single center / 中国胸心血管外科临床杂志

@#Objective    To summarize the clinical experience of thoracoscopic combined subsegmentectomy (CSS). Methods    The clinical data of 76 patients who underwent thoracoscopic CSS in Anqing Municipal Hospital from May 2018 to July 2022 were retrospectively analyzed, including 22 males and 54 females, aged 27.0-76.0 (54.3±10.5) years. All patients underwent preoperative three-dimensional computed tomography bronchography and angiography using dual source CT. The modified inflation-deflation technique or indocyanine green was used to identify the intersubsegmental border. Results    A total of 86 pulmonary nodules were resected in 76 patients. One patient of left upper lobe S1+2c+S4a, 1 patient of right upper lobe S2b+S3a and 1 patient of right upper lobe S1b+S3b were further performed lobectomy due to insufficient margin. One patient of left upper lobe S1+2+S3a was further performed left upper division segmentectomy due to residual atelectasis. One patient of left upper lobe S1+2c+S3a was further performed left upper division segmentectomy due to B3b+c injury, and the rest completed planned surgeries successfully. The operative time was 90.0-350.0 (174.9±53.2) min. The operative hemorrhage volume was 50.0 (20.0, 50.0) mL. The postoperative hospital stay time was 6.0 (5.0, 7.0) d. Postoperative complications included pulmonary infection in 9 patients, hemoptysis in 3 patients, persistent pulmonary leakage>3 d in 4 patients, pneumothorax in 1 patient, pleural effusion in 1 patient, and myocardial infarction in 1 patient. All of the patients were cured and discharged without perioperative death. Conclusion    Thoracoscopic CSS is relatively complex. Preoperative planning under three-dimensional reconstruction and intraoperative fine operation are helpful for safe completion.

Analysis of Knee Joint Stress During Drop Jump Landing in Females with Generalised Joint Hypermobility / 医用生物力学

Objective To analyze the differences of von Mises stress distribution in knee cartilage and meniscus in female with generalised joint hypermobility (GJH) and healthy female during drop jump landing. Methods The kinematic and ground reaction force (GRF) characteristics of knee joint in female with GJH and healthy female at the moment of peak vertical GRF (VGRF) during loading phase of drop jump landing were collected. The knee joint reaction force was calculated via inverse dynamics, and the combined force of knee joint along long axis of the femur was applied as the load. Based on three-dimensional (3D) finite element model of a female knee joint, numerical simulations were performed separately during drop jump landing of subjects in two groups, and von Mises stresses and stress distribution of knee cartilage and meniscus were calculated. Results At the moment of peak VGRF during drop jump landing, knee flexion and valgus angles in GJH group and control group showed a statistical significance (P<0. 05). Compared with control group, knee flexion angle decreased and valgus angle increased in GJH group. During drop jump landing, GJH group bore larger stress inside the knee joint, and stress distribution in weight-bearing areas of the medial and lateral tibiofemoral compartments was uneven, while the lateral femoral cartilage lateral condyle, the anterior and middle lateral of lateral tibial cartilage, the anterior angle and body lateral margin of lateral meniscus were stress concentration sites. Conclusions For females with GJH, the stability of knee joint decreases and force lines change in jumping events, due to the increased range of motion of knee joint and relaxation of joint capsule, which increases the risk of cartilage and meniscal injury in lateral knee joint. During jumping sports, females with GJH should especially prevent knee joint injury caused by altered force lines in frontal plane of knee joint.

Effects of titanium dioxide nanoparticles on circRNA expression profiles in human hepatocellular carcinoma cells HepG2 / 北京大学学报(医学版)

OBJECTIVE@#To investigate the effect of titanium dioxide nanoparticles (TiO2 NPs) on the expression profile of circular ribonucleic acid (circRNA) in human hepatocytes through in vitro cell experiments, and to attempt to understand the potential mechanism of hepatotoxicity through bioinformatics analysis.@*METHODS@#TiO2 NPs were characterized from the aspects of particle size, shape and agglomeration state. The cell counting kit-8 (CCK8) was used to detect the cytotoxicity of TiO2 NPs against human hepatocellular carcinoma cells (HepG2) after exposure to 0, 1.56, 3.13, 6.25, 12.5, 25, 50, 100, and 200 mg/L TiO2 NPs for 24 h or 48 h. The cells were treated at doses of 0 mg/L TiO2 NPs (control group) and 100 mg/L TiO2 NPs (treatment group), and collected after exposure for 48 h, and then RNA from the extracted cell samples was collected and sequenced. The differential circRNAs between the control and the TiO2 NPs treatment groups were screened, and then the enrichment pathway of the differential circRNA target gene was analyzed by multivariate statistics. According to the sequencing results, significantly altered genes and important genes in the significant enrichment pathways were screened, and real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) was performed to verify the results.@*RESULTS@#TiO2 NPs were spherical anatase with a hydrated particle size of (323.50±85.44) nm and a Zeta potential of (-21.00±0.72) mV in a serum-free medium. The results of the CCK8 cytotoxicity assay showed that with the increase of TiO2 NPs concentration, cell viability gradually decreased. A total of 11 478 circRNAs were found by RNA sequencing. Compared with the control groups, TiO2 NPs treatment groups (100 mg/L) had a total of 89 differential circRNAs, of which 59 were up-regulated and 30 were down-regulated. Analysis of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway showed that the targeted genes of differential circRNAs were mainly enriched in fatty acid degradation, Fanconi anemia pathway, and fatty acid metabolism. The expression levels of circRNA.6730, circRNA.3650 and circRNA.4321 were significantly different between the TiO2 NPs treatment group and the control group, which were consistent with the sequencing results.@*CONCLUSION@#TiO2 NPs can induce changes in circRNA expression profile, and epigenetics may play an important role in the mechanism of hepatotoxicity.

Survival of Non-small Cell Lung Cancer Patients Treated with Fuzheng Gushe Therapy： A Real World Study / 中国实验方剂学杂志

ObjectiveTo analyze the long-term survival of non-small cell lung cancer （NSCLC） patients treated with Fuzheng Gushe therapy in the real world. MethodA retrospective cohort study was carried out with the NSCLC patients treated with Fuzheng Gushe therapy from January 2014 to December 2018 in Dongzhimen Hospital， Beijing University of Chinese Medicine as the integrated traditional Chinese medicine （TCM） and western medicine cohort. The NSCLC data from January 2011 to December 2015 in the Surveillance， Epidemiology， and End Results （SEER） database were downloaded as the western medicine cohort. The propensity score matching （PSM） was employed to balance the confounding factors between the groups， and comparable samples were obtained from the two groups. The Log-rank test was conducted to compare the overall survival between the groups and the survival curves were established. ResultThe integrated TCM and western medicine cohort and the western medicine cohort included 511 and 5 022 NSCLC patients， respectively. The age， sex， and chemotherapy had no significant differences between the two cohorts， while pathological type， lymph node metastasis （N）， distant metastasis （M）， surgery， and radiotherapy were different between the two cohorts （P<0.05）. After PSM， 122 comparable samples were obtained. The overall survival analysis showed that the total survival of the integrated TCM and western medicine cohort was better than that of the western medicine cohort （P<0.05）. The stratified survival analysis showed that the integrated TCM and western medicine cohort had survival advantages in females， males， the young， and adenocarcinoma， N+， and M+ patients （P<0.05）. ConclusionFuzheng Gushe therapy can significantly improve the overall survival of NSCLC patients， especially in the females， the males， the young， and the patients with adenocarcinoma， lymph node metastasis， and distant metastasis.

Research Progress of Next Generation Sequencing in Acute Leukemia --Review / 中国实验血液学杂志

With the advent of precision medicine, next-generation sequencing (NGS) is playing an increasingly important role in clinical oncology diagnosis and treatment with its advantages of high sensitivity, high accuracy, high efficiency and operability. NGS reveals the genetic characteristics of acute leukemia(AL) patients by screening for specific disease-causing genes to identify occult as well as complex genetic mutations in patients with AL, leading to early diagnosis and targeted drug therapy for AL patients, as well as to predict disease recurrence by detecting mnimal residual disease (MRD) and analyzing mutated genes to determine patient prognosis. NGS plays an increasingly important role in the diagnosis, treatment and prognosis assessment in AL, providing a direction for the pursuit of precision medicine. This paper reviews the research progress of NGS in AL.

Identification of Long Non-Coding RNAs and Their Target Genes from Mycelium and Primordium in Model Mushroom Schizophyllum commune

Schizophyllum commune has emerged as the most promising model mushroom to study developmental stages (mycelium, primordium), which are two primary processes of fruit body development. Long non-coding RNA (lncRNA) has been proved to participate in fruit development and sex differentiation in fungi. However, potential lncRNAs have not been identified in S. commune from mycelium to primordium developmental stages. In this study, lncRNA-seq was performed in S. commune and 61.56 Gb clean data were generated from mycelium and primordium developmental stages. Furthermore, 191 lncRNAs had been obtained and a total of 49 lncRNAs were classified as differently expressed lncRNAs.Additionally, 26 up-regulated differently expressed lncRNAs and 23 down-regulated between mycelium and primordia libraries were detected. Further, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that differentially expressed lncRNAs target genes from the MAPK pathway, phosphatidylinositol signal, ubiquitin-mediated proteolysis, autophagy, and cell cycle. This study provides a new resource for further research on the relationship between lncRNA and two developmental stages (mycelium, primordium) in S. commune

PathogenTrack and Yeskit: tools for identifying intracellular pathogens from single-cell RNA-sequencing datasets as illustrated by application to COVID-19 / 医学前沿

Pathogenic microbes can induce cellular dysfunction, immune response, and cause infectious disease and other diseases including cancers. However, the cellular distributions of pathogens and their impact on host cells remain rarely explored due to the limited methods. Taking advantage of single-cell RNA-sequencing (scRNA-seq) analysis, we can assess the transcriptomic features at the single-cell level. Still, the tools used to interpret pathogens (such as viruses, bacteria, and fungi) at the single-cell level remain to be explored. Here, we introduced PathogenTrack, a python-based computational pipeline that uses unmapped scRNA-seq data to identify intracellular pathogens at the single-cell level. In addition, we established an R package named Yeskit to import, integrate, analyze, and interpret pathogen abundance and transcriptomic features in host cells. Robustness of these tools has been tested on various real and simulated scRNA-seq datasets. PathogenTrack is competitive to the state-of-the-art tools such as Viral-Track, and the first tools for identifying bacteria at the single-cell level. Using the raw data of bronchoalveolar lavage fluid samples (BALF) from COVID-19 patients in the SRA database, we found the SARS-CoV-2 virus exists in multiple cell types including epithelial cells and macrophages. SARS-CoV-2-positive neutrophils showed increased expression of genes related to type I interferon pathway and antigen presenting module. Additionally, we observed the Haemophilus parahaemolyticus in some macrophage and epithelial cells, indicating a co-infection of the bacterium in some severe cases of COVID-19. The PathogenTrack pipeline and the Yeskit package are publicly available at GitHub.

Study on the diagnostic value of transient elastography, APRI and FIB-4 for liver fibrosis in children with non-alcoholic fatty liver disease / 中华肝脏病杂志

Objective: To evaluate the diagnostic value of transient elastography, aspartate aminotransferase-to-platelet ratio index (APRI), and fibrosis index based on 4 factors (FIB-4) for liver fibrosis in children with non-alcoholic fatty liver disease (NAFLD). Methods: A retrospective study was conducted on 100 cases of nonalcoholic fatty liver disease in Hunan Children's Hospital between August 2015 to October 2020 to collect liver tissue pathological and clinical data. The receiver operating characteristic curve (ROC curve) was used to analyze the diagnostic value of liver stiffness measurement (LSM), APRI and FIB-4 in the diagnosis of different stages of liver fibrosis caused by NAFLD in children. Results: The area under the ROC curve (AUC) value of LSM, APRI and FIB-4 for diagnosing liver fibrosis (S≥1) were 0.701 [95% confidence interval (CI): 0.579 ~ 0.822, P = 0.011], 0.606 (95%CI: 0.436 ~ 0.775, P = 0.182), and 0.568 (95%CI: 0.397 ~ 0.740, P = 0.387), respectively. The best cut-off values were 6.65 kPa, 21.20, and 0.18, respectively. The AUCs value of LSM, APRI, and FIB-4 for diagnosing significant liver fibrosis (S≥ 2) were 0.660 (95% CI: 0.552 ~ 0.768, P = 0.006), 0.578 (95% CI: 0.464 ~ 0.691, P = 0.182) and 0.541 (95% CI: 0.427 ~ 0.655, P = 0.482), respectively. The best cut-off values were 7.35kpa, 24.78 and 0.22, respectively. The AUCs value of LSM, APRI and FIB-4 for the diagnosis of advanced liver fibrosis (S≥ 3) were 0.639 (95% CI: 0.446 ~ 0.832, P = 0.134), 0.613 (95% CI: 0.447 ~ 0.779, P = 0.223) and 0.587 (95% CI: 0.411 ~ 0.764, P = 0.346), respectively. The best cut-off values were 8.55kpa, 26.66 and 0.27, respectively. Conclusion: The transient elastography technique has a better diagnostic value than APRI and FIB-4 for liver fibrosis in children with NAFLD.

Clinical guideline for spinal reconstruction of osteoporotic thoracolumbar fracture in elderly patients (version 2022) / 中华创伤杂志

Osteoporotic thoracolumbar fracture in the elderly will seriously reduce their quality of life and life expectancy. For osteoporotic thoracolumbar fracture in the elderly, spinal reconstruction is necessary, which should comprehensively consider factors such as the physical condition, fracture type, clinical characteristics and osteoporosis degree. While there lacks relevant clinical norms or guidelines on selection of spinal reconstruction strategies. In order to standardize the concept of spinal reconstruction for osteoporotic thoracolumbar fracture in the elderly, based on the principles of scientificity, practicality and progressiveness, the authors formulated the Clinical guideline for spinal reconstruction of osteoporotic thoracolumbar fracture in elderly patients ( version 2022), in which suggestions based on evidence of evidence-based medicine were put forward upon 10 important issues related to the fracture classification, non-operative treatment strategies and surgical treatment strategies in spinal reconstruction after osteoporosis thoracolumbar fracture in the elderly, hoping to provide a reference for clinical treatment.

Percutaneous endoscopic lumbar discectomy in the treatment of adjacent segment lumbar disc herniation after lumbar fusion / 中国骨伤

OBJECTIVE@#To explore the clinical effect of percutaneous endoscopic lumbar discectomy in the treatment of adjacent segment lumbar disc herniation after lumbar fusion.@*METHODS@#From February 2010 to June 2018, 64 patients with adjacent segment lumbar disc herniation after lumbar fusion were retrospectively analyzed and divided into observation group and control group. In observation group, there were 23 males and 10 females performed with percutaneous endoscopic lumbar discectomy, including 27 cases of single segment fusion and 6 cases of double segment fusion, aged from 55 to 83 years old with an average of (65.7±7.4) years old. In control group, there were 22 males and 9 females performed with traditional open fusion revision, including 25 cases of single-segment fusion and 6 cases of double segment fusion, aged from 51 to 78 years old with an average of(64.8±7.8) years old. The operative time, intraoperative blood loss, fluoroscopy times, postoperative ambulation time and length of postoperative hospital stay were recorded. The clinical efficacy was evaluated by visual analogue scale(VAS) and Oswestry Disability Index(ODI). The complications between two groups were observed.@*RESULTS@#All patients were followed up for at least 2 years. The observation group patients were followed up with an average of (2.4±0.5) years. The control group patients were followed up with an average of(2.6±0.7) years. Compared with control group, operation time, intraoperative blood loss, postoperative ambulation time and length of postoperative hospital stay of the observation group were significantly reduced (P<0.05), and the fluoroscopy times of observation group were significantly increased compared with control group(P<0.05). The VAS of low back and lower limb, and ODI at the latest follow-up between two groups were all significantly improved compared to those of pre-operation (P<0.05). The VAS of low back at each point and ODI at 1, 3 months after operation in observation group was significantly reduced compared with control group(P<0.05), however there was no significant difference in VAS for lower limb between two groups (P>0.05). The difference of complications between two groups was statistically significant (P<0.05).@*CONCLUSION@#Compared with traditional open fusion revision surgery, percutaneous endoscopic lumbar discectomy for the treatment of adjacent segment lumbar disc herniation after lumbar fusion has the advantages of reducing operation time and intra-operative blood loss, shortening ambulation time and the length of postoperative hospital stay, and promoting pain and functional improvement, and decrease incidence of complications. However, long-term clinical efficacy needs further study.

A CRISPR/dCpf1-based transcriptional repression system for Gluconobacter oxydans / 生物工程学报

Gluconobacter oxydans are widely used in industrial due to its ability of oxidizing carbohydrate rapidly. However, the limited gene manipulation methods and less of efficient gene editing tools impose restrictions on its application in industrial production. In recent years, the clustered regularly interspaced short palindromic repeats (CRISPR) system has been widely used in genome editing and transcriptional regulation which improves the efficiency of genome editing greatly. Here we constructed a CRISPR/dCpf1-mediated gene transcriptional repression system, the expression of a nuclease inactivation Cpf1 protein (dCpf1) in Gluconobacter oxydans together with a 19 nt direct repeats showed effective repression in gene transcription. This system in single gene repression had strong effect and the relative repression level had been increased to 97.9%. While it could be applied in multiplex gene repression which showed strong repression ability at the same time. Furthermore, this system was used in the metabolic pathway of L-sorbose and the regulatory of respiratory chain. The development of CRISPR transcriptional repression system effectively covered the shortage of current gene regulation methods in G. oxydans and provided an efficient gene manipulation tool for metabolic engineering modification in G. oxydans.