Objective:To explore the effects of Qigui Fuyuan Decoction (QGFYD) on improving neural function, reducing neuronal damage and inflammatory response in rats with cerebral ischemia-reperfusion injury.Methods:A middle cerebral artery occlusion (MCAO) model was established in SD rats using suture occlusion method. The rats were divided into sham-operation group, model group, and QGFYD low- (QGFYD-L), medium- (QGFYD-M), and high-dosage (QGFYD-H) groups according to random number table method. The modified neurological severity scores (mNSS) was used to evaluate the neurological deficit score, Triphenyl tetrazolium chloride (TTC) staining to assess the volume of cerebral infarction, and Western blot for inflammatory factor expression and ELISA assay for serum inflammatory factor levels. Stimulation of PC12 cells with L-glutamic acid (L-Glu) was used to establish an in vitro neurological injury model, and QGFYD-containing serum and intestinal absorptive solution were prepared to study its neuroprotective effects in vitro using the CCK-8 assay kit.Results:Compared with the model group, QGFYD-L, QGFYD-M, and QGFYD-H groups showed a decrease in neurological deficit symptom scores ( P<0.01), a decrease in cerebral infarction volume ( P<0.01), a decrease in serum IL-6 and TNF-α levels ( P<0.05), and a decrease in brain tissue IL-1 β, IL-6, iNOS, and TNF-α protein expression ( P<0.05); both the QGFYD-containing serum and the intestinal absorption solution could alleviate L-Glu induced damage to PC12 cells ( P<0.01). In vitro experiments showed that intervention with 30 mmol/L L-glutamate for 24 hours could cause damage to PC12 cells. Both the serum containing Qigui Fuyuan decoction and intestinal absorption solution could alleviate L-Glu-induced damage to PC12 cells ( P<0.01). Conclusion:Qigui Fuyuan Decoction can alleviate cerebral ischemia-reperfusion injury, improve neurological function, and inhibit neuroinflammation.

Objective To create risk predictive models of healthcare-seeking delay among imported malaria patients in Jiangsu Province based on machine learning algorithms, so as to provide insights into early identification of imported malaria cases in Jiangsu Province. Methods Case investigation, first symptoms and time of initial diagnosis of imported malaria patients in Jiangsu Province in 2019 were captured from Infectious Disease Report Information Management System and Parasitic Disease Prevention and Control Information Management System of Chinese Center for Disease Control and Prevention. The risk predictive models of healthcare-seeking delay among imported malaria patients were created with the back propagation (BP) neural network model, logistic regression model, random forest model and Bayesian model using thirteen factors as independent variables, including occupation, species of malaria parasite, main clinical manifestations, presence of complications, severity of disease, age, duration of residing abroad, frequency of malaria parasite infections abroad, incubation period, level of institution at initial diagnosis, country of origin, number of individuals travelling with patients and way to go abroad, and time of healthcare-seeking delay as a dependent variable. Logistic regression model was visualized using a nomogram, and the nomogram was evaluated using calibration curves. In addition, the efficiency of the four models for prediction of risk of healthcare-seeking delay among imported malaria patients was evaluated using the area under curve (AUC) of receiver operating characteristic curve (ROC). The importance of each characteristic was quantified and attributed by using SHAP to examine the positive and negative effects of the value of each characteristic on the predictive efficiency. Results A total of 244 imported malaria patients were enrolled, including 100 cases (40.98%) with the duration from onset of first symptoms to time of initial diagnosis that exceeded 24 hours. Logistic regression analysis identified a history of malaria parasite infection [odds ratio (OR) = 3.075, 95% confidential interval (CI): (1.597, 5.923)], long incubation period [OR = 1.010, 95% CI: (1.001, 1.018)] and seeking healthcare in provincial or municipal medical facilities [OR = 12.550, 95% CI: (1.158, 135.963)] as risk factors for delay in seeking healthcare among imported malaria cases. BP neural network modeling showed that duration of residing abroad, incubation period and age posed great impacts on delay in healthcare-seek among imported malaria patients. Random forest modeling showed that the top five factors with the greatest impact on healthcare-seeking delay included main clinical manifestations, the way to go abroad, incubation period, duration of residing abroad and age among imported malaria patients, and Bayesian modeling revealed that the top five factors affecting healthcare-seeking delay among imported malaria patients included level of institutions at initial diagnosis, age, country of origin, history of malaria parasite infection and individuals travelling with imported malaria patients. ROC curve analysis showed higher overall performance of the BP neural network model and the logistic regression model for prediction of the risk of healthcare-seeking delay among imported malaria patients (Z = 2.700 to 4.641, all P values < 0.01), with no statistically significant difference in the AUC among four models (Z = 1.209, P > 0.05). The sensitivity (71.00%) and Youden index (43.92%) of the logistic regression model was higher than those of the BP neural network (63.00% and 36.61%, respectively), and the specificity of the BP neural network model (73.61%) was higher than that of the logistic regression model (72.92%). Conclusions Imported malaria cases with long duration of residing abroad, a history of malaria parasite infection, long incubation period, advanced age and seeking healthcare in provincial or municipal medical institutions have a high likelihood of delay in healthcare-seeking in Jiangsu Province. The models created based on the logistic regression and BP neural network show a high efficiency for prediction of the risk of healthcare-seeking among imported malaria patients in Jiangsu Province, which may provide insights into health management of imported malaria patients.

Objective To examine the expression pattern of miR-135a-5p and miR-135b-5p in seminal plasma as well as the clinical value of their variation in the male patients with infertility.Methods The clinical samples of 5 kinds,including serum,morning urine,pleural effusion,cerebrospinal fluid and seminal plasma,were collected from 30 patients respectively in the Department of Clini-cal Laboratory,Jiangsu Province Hospital of Chinese Medicine from January 2019 to December 2021.In addition,the frozen seminal plasma samples from the patients with non-obstructive azoospermia,asthenozoospermia of 54 cases for each illness statuses,and 54 healthy controls with fertile ability from 2010 to 2019 in Jiangsu Province Hospital of Chinese Medicine and Jinling Hospital were also selected.Fluorescent quantitative RT-qPCR analyses were applied to measure the expression levels of miR-135a-5p and miR-135b-5p in seminal plasma.Receiver operating characteristic curve(ROC)was used to construct the area under the curve(AUCROC)to evalu-ate the ability for discriminating infertility males from healthy controls.Correlation analyses were performed to explore the correlations between the levels of miR-135a-5p and miR-135b-5p in seminal plasma and other semen parameters.Results The concentrations of miR-135a-5p and miR-135b-5p in the samples from five different kinds of body fluid showed significantly difference(H=64.88,P＜0.01 and H=64.7,P＜0.01,respectively).The highest levels were found in serum and seminal plasma,and the lowest levels were in u-rine.The concentrations of miR-135a-5p and miR-135b-5p in seminal plasma were also showed marked difference among the azoosper-mia group,asthenozoospermia group and healthy fertile control group(H=32.14,P＜0.01 and H=21.37,P＜0.01,respectively).Compared to the fertile control group,the levels of miR-135a-5p and miR-135b-5p in seminal plasma were significantly downregulated in azoospermia patients(U=687,P＜0.01 and U=843,P＜0.01,respectively).The contents of the two miRNAs showed no statistical-ly difference between asthenozoospermia patients and healthy fertile controls,but showed markedly difference between the two patients groups(U=635,P＜0.01 and U=779,P＜0.01,respectively).Receiver operating characteristic curve(ROC)analyses results re-vealed that levels of miR-135a-5p and miR-135b-5p in seminal plasma could successfully discriminate the azoospermia patients from fertile controls and asthenozoospermia patients with the AUCROC of 0.764(95％CI:0.675 to 0.854),0.711(95％CI:0.612 to 0.810)and 0.782(95％CI:0.695 to 0.869),and 0.733(95％CI:0.637 to 0.829),respectively.Correlation analyses demonstrated that the concentrations of miR-135a-5p and miR-135b-5p in seminal plasma were significantly associated with sperm concentration,total sperm motility,and percentages of progressive sperm and non-progressive sperm(all with the P-value＜0.01).Conclusion miR-135a-5p and miR-135b-5p are the highly expressed miRNA in seminal plasma,and significantly decreased in azoospermia patients.Both of them may play pivot roles in male infertility.

Remote teaching consultation is an online continuing medical education mode which combines medical practice with teaching and superimposes teaching functions on the basis of remote consultation. Based on the pilot experiences of collaboration between Zhongshan Hospital and Xidu Community Health Service Center, this article analyzes the strengths, weaknesses, opportunities and threats (SWOT)of the remote teaching consultation for general practitioners, and discusses strategies to improve the further implementation plan. The analysis showed that as a novel educational method, the remote teaching consultation should take the advantages of online education, make good use of the remote consultation platform, improve its teaching connotation, and form standardized implementation norms to meet the diversified needs of general practitioners for continuing medical education.

Objective: To explore the heterogeneity and correlation of clinical phenotypes and genotypes in children with disorders of sex development (DSD). Methods: A retrospective study of 1 235 patients with clinically proposed DSD in 36 pediatric medical institutions across the country from January 2017 to May 2021. After capturing 277 DSD-related candidate genes, second-generation sequencing was performed to analyzed the heterogeneity and correlation combined with clinical phenotypes. Results: Among 1 235 children with clinically proposed DSD, 980 were males and 255 were females of social gender at the time of initial diagnosis with the age ranged from 1 day of age to 17.92 years. A total of 443 children with pathogenic variants were detected through molecular genetic studies, with a positive detection rate of 35.9%. The most common clinical phenotypes were micropenis (455 cases), hypospadias (321 cases), and cryptorchidism (172 cases) and common mutations detected were in SRD5A2 gene (80 cases), AR gene (53 cases) and CYP21A2 gene (44 cases). Among them, the SRD5A2 mutation is the most common in children with simple micropenis and simple hypospadias, while the AMH mutation is the most common in children with simple cryptorchidism. Conclusions: The SRD5A2 mutation is the most common genetic variant in Chinese children with DSD, and micropenis, cryptorchidism, and hypospadias are the most common clinical phenotypes. Molecular diagnosis can provide clues about the biological basis of DSD, and can also guide clinicians to perform specific clinical examinations. Target sequence capture probes and next-generation sequencing technology can provide effective and economical genetic diagnosis for children with DSD.
3-Oxo-5-alpha-Steroid 4-Dehydrogenase/genetics* ; Child ; China/epidemiology* ; Cryptorchidism/genetics* ; Disorders of Sex Development/genetics* ; Female ; Genital Diseases, Male ; Genotype ; Humans ; Hypospadias/genetics* ; Male ; Membrane Proteins/genetics* ; Penis/abnormalities* ; Phenotype ; Retrospective Studies ; Steroid 21-Hydroxylase/genetics*

The application of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) can be limited due to a lack of compatible protospacer adjacent motif (PAM) sequences in the DNA regions of interest. Recently, SpRY, a variant of Streptococcus pyogenes Cas9 (SpCas9), was reported, which nearly completely fulfils the PAM requirement. Meanwhile, PAMs for SpRY have not been well addressed. In our previous study, we developed the PAM Definition by Observable Sequence Excision (PAM-DOSE) and green fluorescent protein (GFP)‍-reporter systems to study PAMs in human cells. Herein, we endeavored to identify the PAMs of SpRY with these two methods. The results indicated that 5'-NRN-3', 5'-NTA-3', and 5'-NCK-3' could be considered as canonical PAMs. 5'-NCA-3' and 5'-NTK-3' may serve as non-priority PAMs. At the same time, PAM of 5'-NYC-3' is not recommended for human cells. These findings provide further insights into the application of SpRY for human genome editing.
CRISPR-Associated Protein 9/metabolism* ; CRISPR-Cas Systems ; DNA ; Gene Editing/methods* ; Humans ; Streptococcus pyogenes/metabolism*

Bronchiolitis is the most common lower respiratory tract infection and the leading cause of hospitalization in infants. It has become one of the public health problems worldwide. The pathogenesis is unclear and there are currently no targeted therapies that have been proven to be effective in bronchiolitis. There exists a poor understanding of the mechanisms underlying the variable disease severity and the inconsistent manifestation of long-term respiratory morbidity. Nor can these outcomes be reliably predicted in patients on initial presentation. Bronchiolitis is generally considered a single disease,however,more and more researches have shown that it is a heterogeneous disease that needs individualized and specific treatment,with differences in etiology,clinical charicteristics,immunology,microecology and long-term respiratory outcomes. The study on heterogeneity of viral bronchiolitis will contribute to individualized and targeted therapy and may reduce the development of recurrent wheezing and asthma.

Objective To compare ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) and chemiluminescence immunoassay(CLIA) measurement of human serum androgen levels in polycystic ovarian syndrome(PCOS). Methods 160 PCOS patients and 146 healthy subjects(control group) were recruited and their serum androgen levels——measurements of testosterone and dehydcoepiandrosterone sulfate (DHEA-S) were tested by UPLC-MS/MS and CLIA. The androgen results combined with body mass index(BMI) were analyzed by ROC curve. According to area under curve(AUC) calculated by SPSS, a better method will be selected to provide accurate test results for physicians. Results (1)AUC of DHEA-S tested by UPLC-MS/MS was significantly higher than the one tested by CLIA(P<0.01). There was no significant difference between the AUC of T tested by UPLC-MS/MS and the one tested by CLIA. (2)AUC of T combined with DHEA-S tested by HPLC was not only significantly higher than the AUC of two combined indicators tested by CLIA(P<0.01),but also significantly higher than the AUC of a single indicator——either T(P<0.01) or DHEA-S(P<0.01) tested by UPLC-MS/MS. (3)The AUC of T,DHEA-S combined with BMI tested by HPLC was not only significantly higher than the AUC of three combined indicators tested by CLIA(P<0.01),but also higher than the AUC of two combined indicators tested by UPLC-MS/MS(P<0.05). Conclusion T and DHEA-S tested by UPLC-MS/MS combined with BMI has a certain reference value in the diagnosis of PCOS.

Objective To investigate the CT manifestations and pathology features of solitary fibrous tumors of the pleural (SFTP),to improve diagnostic accuracy of this disease.Methods CT manifestations of 12 cases with SFTP confirmed by surgery were analyzed retrospectively by comparison with the pathological findings.Results The CT images of the 12 cases showed intrathoracic soft-tissue shadow with well-defined margin which was widely connected with pleural.There were 4 cases with mound like and mushroom shape,3 cases with round or oval shape,2 cases with irregular shape growing into the pleural space,3 cases with large mass filling unilateral thoracic cavity,9 cases with pleural tail sign and 1 case with pleural pedicle sign.The plain CT showed homogeneous density in 7 cases,high-low mixed density in 4 cases and with internal calcification in 1 case.After contrast enhancement,5 cases were map like heterogeneous and progressive enhancement,and 4 cases have strip distorted vessel inside or on the edge of the lesion.Conclusion CT manifestations of SFTP have certain characteristics,such as broad base,shaping growth and pleural tail sign,which could be helpful to improve the diagnostic accuracy of SFTP by analyzing carefully.

Objective To investigate the status of health promotion,influencing factors,and health promotion level of cancer patients with PICC during tube indwelling period.Methods A total of 125 PICC cancer patients were investigated by using general information questionnaire and Health Promoting Lifestyle Scale (HPLP-Ⅱ R) according to convenience sampling.Results The average score of health promotion lifestyle in cancer patients was(100.70 ± 18.42)scores during PICC,and the overall level was at the middle level.There was a negative correlation between health promotion behavior and age in cancer patients during PICC detention (P ＜ 0.01),and positively correlated with education,family income and catheterization days (P ＜ 0.05).Multivariate analysis showed that age,education level,diagnosis and catheterization days were influencing factor of health promotion behavior level of cancer patients during PICC indwelling.Conclusion The health promotion behavior of cancer patients during PICC stay is in general level.In clinical nursing work,these patients with elder age,low education level and short timne of catheterization should be paid much attention,so as to improve their health promoting behavior.