Selection markers are essential tools in gene editing, the utility of such systems is inherently constrained by species-specific limitations, governed by divergent host genetic backgrounds and metabolic compatibility. To address this limitation, we leveraged the CRISPR/Cas9 system to develop a universal counter-selection tool. We designed and introduced an sgRNA expression cassettes as counter-selection markers, which directs the Cas9 protein to target and cleave genomic DNA, allowing for the selection of the strains where the sgRNA expression cassette has been replaced. Optimized to target multiple copy sites with sgRNA, this system significantly enhances cell lethality, boosting counter-selection efficiency to over 85.00%. This counter-selection tool is not limited to single strains and is suitable for various scenarios, including multi-copy plasmid assembly and plasmid editing, demonstrating broad application potential.
CRISPR-Cas Systems/genetics* ; Gene Editing/methods* ; RNA, Guide, CRISPR-Cas Systems/genetics* ; Plasmids/genetics*

AIM: To investigate the effects of hypobaric hypoxia in plateau on tear indexes and related anatomical structures in rabbits.METHODS: A total of 18 healthy New Zealand rabbits were selected and randomly divided into plateau group and control group, with 9 rabbits(18 eyes)in each group. The plateau group was housed in the Simulated Climate Cabin for Special Environment of Northwest of China, simulating hypobaric hypoxia at an altitude of 6 000 m. The control group was housed in a clean animal room with atmospheric pressure and oxygen. Changes in the tear meniscus height and non-invasive tear break-up time were detected by using RHCT-1 corneal topographer dry eye comprehensive analysis system, changes in tear secretion was measured by Schirmer Ⅰ test, before intervention and on the 3, 7 d, 2 and 4 wk. Meanwhile, the changes in tear composition before and after intervention in the plateau environment were analyzed using Raman Spectroscopy. The histopathological changes of the lower lid conjunctiva, cornea, lacrimal gland, and Hardarian gland were observed by hematoxylin-eosin(HE)staining after 4 wk of intervention, and the expression of mucin 5AC(MUC5AC)in conjunctiva was detected by immunohistochemistry.RESULTS: Compared with the control group, Schirmer Ⅰ test, tear meniscus height, first and average non-invasive tear break-up time in the plateau group decreased significantly since 3 d, and the difference was significant with the extension of observation time(P<0.05). The above indexes increased from 2 wk. After 4 wk of intervention, the protein and lipid content of the tear composition of rabbits in the plateau group increased, and the nucleic acid content decreased compared with the pre-intervention period. Compared with the control group, rabbits in the plateau group showed thickening of corneal stromal edema, an increase in the number of conjunctival cup cells, increase in the level of expression of MUC5AC, an increase in the level of expression of MUC5AC, an atrophy and flattening of cytoplasm in lacrimal epithelial cells, an enlargement of glandular lumen, and no obvious destructive changes in the Hardarian glands.CONCLUSION: Acute plateau environment can destroy the homeostasis of rabbit ocular surface, so that the tear secretion and the tear film stability decreases, but within a certain period of time, rabbits undergo compensation with the habituation to the hypobaric hypoxia environment, which can increase the tear secretion to a certain extent and restore the tear film stability.

ObjectiveTo analyze the research status and trends in low-level occupational benzene exposure. Methods Articles on low-level occupational benzene exposure from Chinese and English journals from January 1st, 2000, to December 31th, 2022 were retrieved using the Web of Science and the China National Knowledge Infrastructure, and a bibliometric analysis was conducted. Results A total of 327 articles were included in the analysis, comprising 216 English articles and 111 Chinese articles. i) The number of articles published in English fluctuates greatly over the years, without a trend of continuous growth or decline. Authors from 359 research institutions in 45 countries and regions have published relevant English articles in 97 kinds of journals, involving 281 grants from 226 foundations. The top three countries in terms of articles amount were the United States, Italy, and China, with 81, 46, and 43 papers, respectively. The English articles mainly focused on mechanistic research at the genetic level, such as hematotoxicity, oxidative stress, and DNA damage. ii) The number of Chinese articles increased gradually after 2012, with the growth peak in 2017. Authors from 127 research institutions in 26 provinces, autonomous regions, and municipalities published Chinese articles in 51 kinds of journals, involving 154 grants from 78 foundations. Chinese articles tended to focus on benzene-induced hematotoxicity and occupational health damage. Conclusion Most studies on low-level occupational benzene exposure were conducted in China, the United States and Italy, focused on hematotoxicity. Monitoring international research topics and hotspots of the field has certain reference value for related research in China.

Frailty is a geriatric syndrome associated with aging, which is characterized by a decrease in the ability to stress and maintain homeostasis after the loss of physiological reserve function, resulting in an increased risk of falls, disability, and even death.Cognitive impairment is a disorder of cognitive function due to a variety of causes, between normal aging and dementia.In the early stages, frailty is a certain reversibility.As the core of physical frailty, sarcopenia has been a research hotspot in recent years.With the deep research on sarcopenia, researchers have found that is closely related to cognitive disorder.Muscle-brain axis may play a role in sarcopenia and cognitive impairment.This article illustrates its possible mechanisms at the crossroad of sarcopenia and cognitive disorder, and to provide reference for exploring new targets for intervention of sarcopenia and cognitive impairment.

Objective:To analyze the influential factors of blurred vision after general anesthesia.Methods:The clinical data of 997 patients who underwent elective general anesthesia at The No. 1 People's Hospital of Pinghu from September 2022 to May 2023 were retrospectively analyzed. The data collected included age, sex, body mass index, American Society of Anesthesiologists classification, history of hypertension, history of diabetes, operation duration (specifically whether it exceeded 3 hours), surgical position (whether the patient was in the supine position), operating room temperature, use of penehyclidine hydrochloride, use of muscle relaxant antagonists, use of atropine, blood pressure (specifically whether it was ≥ 30% of the baseline value), fluid input, blood loss, and use of pneumoperitoneum. Collinearity diagnosis and univariate logistic regression analysis were conducted to select factors with statistical significance. Subsequently, multivariate logistic regression analysis was performed.Results:Univariate and multivariate logistic regression analyses showed that age > 65 years ( OR = 1.47, 95% CI: 1.01-2.15, P = 0.043), surgical position (non-supine position) ( OR = 1.54, 95% CI: 1.06-2.25, P = 0.025), operation time exceeding 3 hours ( OR = 1.76, 95% CI: 1.05-2.94, P = 0.031), and the use of penehyclidine hydrochloride ( OR = 4.91, 95% CI: 3.35-7.21, P < 0.001) were identified as factors contributing to postoperative blurred vision in patients undergoing general anesthesia. Conclusion:Factors contributing to postoperative blurred vision in patients undergoing general anesthesia include age > 65 years, the use of penehyclidine hydrochloride during surgery, operation time exceeding 3 hours, and non-supine surgical position. Clinically, it is essential to implement early and effective preoperative education, enhance intraoperative nursing quality, and optimize preoperative medication for general anesthesia to reduce the incidence of blurred vision after surgery.

ObjectiveTo study the efficacy of community rehabilitation combined with drug therapy on rehabilitation of patients with schizophrenia in rural communities， and to provide references for community rehabilitation of patients with schizophrenia in rural areas of our country. MethodA total of 81 patients in rural communities of three towns in Lanzhou new area who met the diagnostic criteria of International Classification of Diseases， tenth edition （ICD-10） were randomly divided into study group （n=39） and control group （n=42）. Both groups received general drug treatment. On this basis， the study group received community rehabilitation intervention for 6 months. Before and after intervention， Positive and Negative Syndrome Scale （PANSS）， Activity of Daily Living Scale （ADL）， Social Disability Screening Schedule （SDSS） and Schizophrenia Quality of Life Scale （SQLS） were used to assess the psychotic symptoms， social function and quality of life in two groups. ResultsAfter intervention， the PANSS total score ［（55.54±14.75） vs. （63.52±13.95）， t=-2.504， P=0.014］， negative symptom factor score ［（15.64±4.50） vs. （18.38±5.13）， t=-2.547， P=0.013］ and general psychopathological factor score ［（25.67±7.39） vs. （30.35±6.60）， t=-3.015， P=0.003］ of the study group were lower than those of the control group. The SDSS score ［（8.21±3.78） vs. （10.21±4.67）， t=-2.118， P=0.037］ and SQLS score ［（18.97±6.23） vs. （22.43±8.04）， t=2.150， P=0.035］ of the study group were lower than those of the control group. ConclusionCommunity rehabilitation combined with drug therapy may help alleviate psychotic symptoms， improve social function and improve quality of life in patients with schizophrenia in rural communities.

The current document is based on a consensus reached by a panel of experts from the Chinese Society of Allergy and the Chinese Society of Otorhinolaryngology-Head and Neck Surgery, Rhinology Group. Chronic rhinosinusitis (CRS) affects approximately 8% of Chinese adults. The inflammatory and remodeling mechanisms of CRS in the Chinese population differ from those observed in the populations of European descent. Recently, precision medicine has been used to treat inflammation by targeting key biomarkers that are involved in the process. However, there are no CRS guidelines or a consensus available from China that can be shared with the international academia. The guidelines presented in this paper cover the epidemiology, economic burden, genetics and epigenetics, mechanisms, phenotypes and endotypes, diagnosis and differential diagnosis, management, and the current status of CRS in China. These guidelines—with a focus on China—will improve the abilities of clinical and medical staff during the treatment of CRS. Additionally, they will help international agencies in improving the verification of CRS endotypes, mapping of eosinophilic shifts, the identification of suitable biomarkers for endotyping, and predicting responses to therapies. In conclusion, these guidelines will help select therapies, such as pharmacotherapy, surgical approaches and innovative biotherapeutics, which are tailored to each of the individual CRS endotypes.
Adult ; Asian Continental Ancestry Group ; Biomarkers ; China ; Consensus ; Diagnosis ; Diagnosis, Differential ; Drug Therapy ; Eosinophils ; Epidemiology ; Epigenomics ; Genetics ; Humans ; Hypersensitivity ; Inflammation ; International Agencies ; Medical Staff ; Neck ; Phenotype ; Precision Medicine

Objective:To explore influence on physical development of children aged 18 months from HIV-positive mothers for prevention mother to child transmission of HIV (PMTCT) in Guangxi Zhuang autonomous region, and provide evidence for the improvement PMTCT program.Methods:This retrospective case control study was conducted in 554 HIV negative infants aged 18 months whose HIV positive mothers had received PMTCT services reported through PMTCT system database from January 1, 2010 to December 31, 2017 and 1 109 healthy infants born in 2017, whose mothers were healthy, in Lingshan, Luzhai, and Hengxian counties, ranking top three counties with high HIV infection prevalence, in Guangxi. PMTCT data and physical development data such as height, weight and head circumference of children aged 18 months were collected. The physical dysplasia in the infants was defined as at least one of the three main indicators of height, weight and head circumference below the normal range.Results:The number of HIV-positive mother and their infants in the case group were 667 and 554 respectively, and the PMTCT rates were 91.15 % (608/667) and 96.57 % (535/554) respectively. HIV positive rate, mortality rate and mother to child transmission rate of the infants aged 18 months were 1.44 % (8/554), 3.07 % (17/554) and 1.91 % (8/418) respectively, and the physical examination results of the infants aged 18 months showed that the physical dysplasia rate was 30.51 % (169/554). Among the 1 109 infants in the control group, the physical dysplasia rate was 9.83 % (109/1 109). The difference between the case group and the control group was significant ( P<0.01). Conclusion:The PMTCT rates of HIV positive mother and their children were more than 90.00 %, respectively. However, poor physical development rate of infants aged 18 months were more than 30.00 %. The possible influence of PMTCT on physical development of the infants aged 18 months of HIV positive mother’s needs to be further studied.

Objective To explore the effect of oral-muscle biofeedback system training on the rehabilitation of cognitive dysfunction in patients with obstructive sleep apnea (OSA) after ischemic stroke.Methods A total of 60 cases of poststroke OSA patients with cognitive dysfunction admitted to the rehabilitation center of deAn hospital in changzhou city from June 2017 to March 2018 were selected.On the basis of routine rehabilitation treatment,the control group was given positive pressure ventilation treatment,and the observation group was given oral muscle biofeedback system training.The changes of polysomnography (PSG) parameters,Montreal cognitive assessment (MoCA) score,serum matrix metalloproteinase-9 (MMP-9) and plasma homcysteinemia (Hcy) expression levels in the two groups before and after treatment were compared.Results Apnea hypopnea index and maximum apnea time of the two groups were significantly decreased after treatment compared with those before treatment,and the lowest SpO2 and average SpO2 were significantly higher than those before treatment(P＜0.05).After treatment,MoCA total score and the dimensions score of patients in the observation group were significantly higher than those in the control group at the same time (P＜0.05).After treatment,the levels of Hcy((12.38±4.54) μmol/L) and MMP-9((124.52± 12.94) μg/L) in the observation group were significantly lower than those in the control group at the same time(Hcy(17.52±6.27) μmol/L,MMP-9(268.25±20.5 7)μg/L),and the difference was statistically significant (Hcy:t=3.637,P＜0.05;MMP-9:t =32.395,P＜0.05).Conclusions Positive pressure ventilation and oral muscle biofeedback system training have the same effect on OSA after stroke,but oral muscle biofeedback system training can also effectively improve patients' cognitive function.

Objective To explore the application disturbance coefficient (DC) value of noninvasive brain edema monitoring in patients after traumatic brain injury.Methods A total of 54 cerebral injury patients were performed by non-invasive brain edema monitoring from June to November 2016.The essential information,DC,intracranial pressure (ICP),and 6-month-later glasgow outcome score (GOS) were collected.Results DC was negatively correlated with ICP (r=-0.779 5,P＜0.01),and it was positively correlated with glasgow coma scale (GCS) and GOS (r=0.667 5,P＜0.01;r=0.630 6,P＜0.01).The mean of DC with good prognosis patients was 106.99±4.09,and that of the poor prognosis patients was85.26±4.45,the difference was statistically significant (P＜0.05).Conclusion DC has a good clinical application value.