OBJECTIVETo study the roles of rhoassociated coiled coil forming protein kinase 2 (Rock2) and transforming growth factor-β1 (TGF-β1) mRNA in acute asthma and the effect of glucocorticoid intervention on the Rock2 and TGF-β1 mRNA expression in rats.

METHODSForty-eight male rats were randomly divided into 4 groups (n=12 each): asthma, control, dexamethasone treated (DXM) and budesonide treated (BUD). Rat model of asthma was prepared by the ovalbumin (OVA) challenge. The animals were sacrificed 24 hrs after the last challenge. The total cell number and differentiation cell number were counted in bronchoalveolar lavage fluid (BALF). The protein expression of Rock2 was ascertained by immunohistochemistry and the mRNA expression of TGF-β1 was ascertained by hybridization in situ.

RESULTSThe pathological changes in the BUD and the DXM groups were alleviated when compared with the asthma group. The total cell number and the percentage of eosinophil (EOS), polymorphonuclear leukocytes (PMN) and lymphocytes (Lym) in BALF in the asthma group were significantly higher than those in the control group (P<0.01). The percentage of macrophage (Mф) in the asthma group was significantly lower than that in the control group (P<0.01). The total cell number and the percentage of EOS and Lym in BALF in the DXM and the BUD groups decreased, while the percentage of Mф increased significantly compared with those in the asthma group (P<0.01). The Rock2 and TGF-β1 mRNA expression in lung tissues in the asthma group increased significantly compared with those in the control, BUD and DXM groups, while there were no significant differences in the Rock2 expression and TGF-β1 mRNA expression between the DXM or BUD group and the control group.

CONCLUSIONSThe expression of Rock2 and TGF-β1 mRNA in lung tissues is increased in rats with acute asthma. Glucocorticoids can significantly decrease the expression of Rock2 and TGF-β1 in lung tissues, thus alleviates airway inflammation.

Animals ; Asthma ; drug therapy ; metabolism ; Budesonide ; therapeutic use ; Dexamethasone ; therapeutic use ; Glucocorticoids ; therapeutic use ; Lung ; metabolism ; pathology ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; genetics ; rho-Associated Kinases ; genetics

Objective:To preliminarily observe the clinical efficacy of microwave hyperthermia combined with intensity-modulated radiotherapy (IMRT) and chemotherapy for patients with locally advanced gastric cancer.Methods:Forty patients who could not been operated or refused operation were enrolled in this clinical trial, who were confirmed as locally advanced proximal or distal gastric cancer by gastroscopy pathology and imaging. Radiotherapy was delivered by IMRT technology for 5 times per week with a total dose of 46 to 56 Gy (median dose of 50 Gy) in 25 to 28 fractions. Synchronous hyperthermia was given at 42 to 44℃ twice a week, 45 min/time. S-1 or capecitabine-based synchronous chemotherapy was performed, d1-14/3 weeks. The symptom remission rate, adverse reactions, objective remission rate (complete and partial remission) and survival were observed.Results:A total of 40 patients, aged between 56 and 83 years (median age of 71 years), were enrolled in this study. The male-to-female ratio was 7: 1. Among them, 38 cases (95%) showed symptom remission. The most common adverse reactions were grade 1-2 gastrointestinal reactions and leukopenia. The objective remission rate was 87.5%, the 2-year progression-free survival and overall survival rates were 68.6% and 70.5%, respectively.Conclusion:Preliminary findings demonstrate that microwave hyperthermia combined with chemoradiotherapy achieve satisfactory outcomes and yield tolerable toxicity in patients with locally advanced gastric cancer.

Objective:To develop the Caregiver Participation Behavior Questionnaire for Elderly Stroke Home Health Management and conduct psychometric evaluation.Methods:Based on the WHO policy framework of active aging, an initial questionnaire pool was constructed through review extensive literature. The initial questionnaire was revised through expert meeting, pilot survey and semantic debugging of the subject. Using multi-stage sampling, 336 caregivers of elderly stroke patients at home in Shanghai were selected from July 2021 to January 2022 to conduct a survey and test the reliability and validity of the questionnaire, and finally a formal questionnaire was formed. A total of 336 questionnaires were distributed. After removing 31 invalid questionnaires that were dropped out of the survey due to personal reasons of some subjects and missing answers, 305 valid questionnaires were recovered, with a valid recovery rate of 90.77% (305/336) .Results:The formal questionnaire included 6 dimensions and 33 items. Exploratory factor analysis extracted 6 common factors, including self-cognition adjustment, community resource utilization, lifestyle change, disease symptom treatment, psychological support, and adherence to rehabilitation exercise. The cumulative variance contribution rate was 81.06%. The item level content validity index was 0.827 to 1.000, and the scale level content validity index was 0.917. The total questionnaire Cronbach's α was 0.887, the split half reliability coefficient was 0.839, and the retest reliability coefficient was 0.812.Conclusions:The reliability and validity of the questionnaire are in line with the statistical test standard, which can be used as a reliable tool to evaluate caregivers' participation behavior in the elderly stroke home health management.

Objective:To investigate the CT and MRI features of sinonasal lymphoepithelial carcinoma (LEC) and to compare functional MRI index of LEC with olfactory neuroblastoma (ONB) in this area.Methods:The clinical data, CT and MRI conventional findings of 7 patients were retrospectively reviewed . All of the patients, 5 males and 2 females, with age of 35-58 (45±9) years old, were histologically diagnosed as LEC of sinonasal tract at Beijing Tongren Hospital, Capital Medical University from February 2014 to November 2019. The semi-quantitative DCE-MRI parameters and ADC value were measured and compared with 18 cases of ONB which were collected during the same period. Independent sample t test or corrected t test, Fisher′s exact test were performed to compare the differences between the two groups. Results:In all of the 7 LEC patients, 6 lesions were located in naso-ethmoid area, 1 lesion was located in naso-orbital area. Six lesions were on the left side, and another one was sitted on both sides. All tumors manifested irregular soft tissue density with bony destruction, 3 of them showed bony sclerosis. All lesions showed homogeneous density or signal and obvious enhancement in solid portion accompanied with varying degrees of local invasion. Morphologically, two forms were observed: localized soft tissue mass and diffuse infiltration along mucosa. There were polypoid strips ( n=3), spherical masses ( n=3), and diffuse thickening of nasal cavity and turbinate mucosa ( n=1). DCE-MRI examinations were performed in six LEC patients. Maximum contrast enhancement index (CI max) was 1.51±0.46, wash out ratio (WR) was 14.26%±6.24%. Time intensity curve (TIC) showed a plateau pattern (type Ⅱ) in 4 cases and a washout pattern (type Ⅲ) in 2 cases. Time to peak (T peak) were (40.09±4.59)s, showing a rapid peak performance. Diffusion weighted imaging was performed in six LEC patients, and the ADC value was (0.80±0.14)×10 -3 mm 2/s. There were statistical differences in WR and ADC values between sinonasal LEC and ONB ( t=4.873, P=0.020 and t=3.255, P=0.025). Conclusion:The radiological manifestations of LEC have certain characteristics. The characteristics of conventional imaging findings and functional MRI index combined with clinical data are helpful in making differential diagnosis between LEC and other sinonasal malignant tumors.

Objective:To understand the current status of nursing for chronic constipation patients accepted fecal microbiota transplantation and provide reference basis for constructing clinical nursing plan.Methods:From April to August 2021, a field research was conducted in the Tenth People′s Hospital of Tongji University. Data was collected by field observation and informal interview for 13 nurses and analyzed by three-level coding method of qualitative research.Results:The work content of the observation subjects could be divided into 3 items including entrance health education, donor management, bacterial fluid management and clinical nursing. It still needed being improved in donor management, health education, nursing of naso-jejunal tube, intestinal preparation, infusion of bacterial fluid, observation of complications and follow-up.Conclusions:It still needs further development in nursing for chronic constipation with fecal microbiota transplantation. It is urgent to establish donor follow-up team, conduct professional training for nurses, rely on mobile medical platform to improve quality of fecal microbiota transplantation, so as to promote the recovery of patients.

Objective:Using clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein 9 (Cas9) technology to edit Vaccinia virus (VACV) thymidine kinase (TK) Region for targeted recombination to establish an efficient vaccinia virus insertion recombination technology.Methods:We designed and synthesized guide RNAs(gRNA)targeting the TK region and then cloned individual gRNA into the PX458 vector that removes nuclear localization signals, and modified the original TK region recombinant plasmid. The gRNA and Cas9 co-expression plasmids were transfected into 293T cells separately to mediate the homologous recombination of vaccinia virus (VACV) and TK region recombination plasmid, and then the rate of viral recombination was evaluated by the appearance of blue and white spots.Results:The recombination efficiency mediated by the gRNA sequence designed and synthesized in this study is more than 1%, which is more than 10 times higher than the classical homologous recombination method .Conclusions:This study used CRISPR/Cas9 technology to establish a highly efficient recombinant vaccinia virus system, which provides technical support for pre-clinical research in vaccines or multivalent research of emerging infectious diseases, as well as tumor treatment.

Objective:To evaluate the immunogenicity of a novel influenza virus mRNA vaccine based on conserved antigens delivered by lipopolyplex (LPP) platform in a mouse model.Methods:Four copies of genes coding for extracellular domain of matrix 2 protein (M2e) and nucleoprotein (NP) of influenza A virus were synthetized after codon optimization. The fusion antigens were transcribed in vitro and delivered by LPP platform, named as LPP-4M2eNP. Expression of M2e and NP in eukaryotic cells was detected by immunofluorescence assay (IFA). BALB/c mice were inoculated intramuscularly twice with 10 μg or 30 μg LPP-4M2eNP vaccine at an interval of four weeks. Antibody response was detected by ELISA and cellular-mediated immunity (CMI) was detected by enzyme-linked immunospot assay (ELISPOT). Results:IFA showed that NP and M2e were expressed correctly in eukaryotic cells. Single dose immunization could induce significant antigen (NP, M2e)-specific CMI and antigen (NP, M2e)-specific antibody response was induced in mice with Th1 type bias after boost immunization. Moreover, NP-specific CMI was increased significantly after the second immunization, while no significant change in M2e-specific CMI was observed.Conclusions:Stronger CMI was triggered in mice by single dose of LPP-4M2eNP vaccine. Furthermore, robust humoral and cellular immune responses were induced after boost immunization. This study suggested that LPP-4M2eNP vaccine, which based on conserved antigen of influenza A and delivered by LPP platform, had great potential for development and application.

Objective:To improve the consistency of test results through reducing inter-laboratory variation in SARS-CoV-2 antibody detection with WHO SARS-CoV-2 antibody candidate international standard (IS, sample G) and antibody reference panel (samples E, F, H, I, J).Methods:Ten WHO samples (A-J) including the candidate IS and reference panel were evaluated using different methods, such as microneutralization tests based on live SARS-CoV-2, pseudovirus neutralization assay and commercial ELISA kits. The test results were compared using statistical analysis.Results:Using IS (sample G) as a reference, the relative concentrations of other samples could be determined with less variation. ELISA and pseudovirus neutralization assay had consistent results with those obtained with the microneutralization test based on SARS-COV-2 strain HB02. Weakly positive samples could be detected only by a certain kit.Conclusions:The availability of an IS for antibodies would facilitate the standardization of SARS-CoV-2 antibody detection methods. The reference panel fitted all the assays based on the SARS-CoV-2 prototype Wuhan strain. Pseudovirus neutralization assay and ELISA could be used as alternatives to live SARS-CoV-2-based neutralization test to some extent.

Objective:To develop a recombinase-aided amplification (RAA)-clustered regularly interspaced short palindromic repeats(CRISPR)/Cas12a-based nucleic acid assay for monkeypox virus with high specificity and sensitivity.Methods:RAA primers and CRISPR RNA (crRNA) were designed based on the known conserved regions of the monkeypox virus gene and synthesized, and specific crRNAs were screened using fluorescence detection. The sensitivity and specificity of the detection system were evaluated.Results:An RAA-CRISPR/Cas12a-based nucleic acid assay for monkeypox virus was developed with a sensitivity of 2.5 copies/reaction and high specificity without cross-reactivity with ectromelia virus and vaccinia virus.Conclusions:An RAA-CRISPR/Cas12a-based nucleic acid assay for monkeypox virus was established, which would provide a powerful tool for efficient, rapid and specific detection of monkeypox virus.

OBJECTIVE: To investigate the effects and safety of catgut embedding on alleviating insomnia. METHODS: Totally 510 patients with insomnia were divided into 5 Chinese medicine (CM) syndrome types: Xin (Heart) and Pi (Spleen) deficiency, yin deficiency with excess fire, Xin and gut qi deficiency, Wei (Stomach) disorder, and qi and blood deficiency, respectively. These 5 types of patients were randomly assigned to a catgut embedding group, an acupuncture group or a medication group (30 cases in Xin and Pi deficiency type, Wei disorder type, Xin and gut qi deficiency type, respectively; 40 cases in yin deficiency with excess fire type and qi and blood deficiency type, respectively). In the catgut embedding group, patients were treated by implanting catgut into acupoints once every 10 days for a total of 30 days. In the acupuncture group, patients were treated with acupuncture once per day over 30 days (excluding weekends); and patients in the medication group took 1 mg Eurodin Tablet orally every night for 30 days. Pittsburgh Sleep Quality Index (PSQI) was evaluated before treatment, on 30 and 60 days after the first treatment, respectively. The International Unified Sleep Efficiency Value (IUSEV) was measured at 30 and 60 days. The safety was evaluated after treatment and adverse events were analyzed. RESULTS: The objective PSQI scores including subjective sleep quality, sleep latency, sleep duration, habitual sleep efficiency, sleep disturbance, daytime dysfunction, and total scores at 30 days were significantly improved compared with pre-treatment in the catgut embedding and acupuncture groups (P<0.01 or P<0.05). At 30 days, the PSQI scores in catgut embedding group were superior to the medication group in the patients with each type of insomnia, with the exception of sleep duration (P<0.01 or P<0.05). At 60 days, significant differences were found between the catgut embedding group and the medication group (P<0.01 for all indices). The IUSEV scores in the catgut embedding group were significantly higher than the acupuncture group at 60 days, and the scores in acupuncture group were higher than the medication group at 30 days (P<0.05 for all types). No severe adverse events were found in this study. CONCLUSIONS Acupoint catgut embedding and acupuncture were more effective than medication in alleviating insomnia syndrome in different Chinese medicine syndrome type. However, the sustained effects of acupoint catgut embedding were superior to acupuncture.