Objective:To explore the feasibility of inguinal subcutaneous immunotherapy for allergic rhinitis ( AR ) in mice. Methods:36 female BALB/c mice were divided randomly into six groups( n=6 per group) including the control A,the model A, the treatment A groups,and the control B,the model B,the treatment B groups(inguinal subcutaneous immunotherapy for group A, cervical back subcutaneous immunotherapy for for group B). AR model was established with ovalbumin. At 25 to 55 days,ovalbumin im-munotherapy were performed in treatment groups,once two days,15 times totally. After intranasal rechallenge was performed at 56 to 62 days the AR symptom scores were documented. The eosinophils(EOS)in the nasal mucosa were measured by chromotropic acid 2R staining. Ovalbumin-specific IgE( OVA-sIgE) in the serum and expression of interferon-γ and interleukin-4 in the nasal lavage were measured by enzyme-linked immunosorbent assay meanwhile the ratio of interferon-γ and interleukin-4 was calc μlated. SPSS17. 0 software was used to analyze the data. Results:Before treatment ,the AR symptom scores of the model and treatment groups were more than 5. After treatment,the treatment A group were less than 5. The EOS count of the control A,model A,treatment A groups and the control B,model B, treatment B groups was 0. 78 ± 0. 31, 21. 60 ± 2. 90, 10. 43 ± 2. 56, 0. 83 ± 0. 46, 22. 44 ± 3. 39, 23. 40 ± 4. 24, respectively. The EOS count of the treatment A group was significantly lower than those in model A group ( P<0. 05 ) . There was no significant difference between treatment B and model B group ( P>0. 05 ) . OVA-sIgE expressed was negative in control groups and positive in other groups. The ratio of interferon-γ and interleukin-4 was 10. 75 ± 3. 38,10. 38 ± 3. 08,3. 02 ± 0. 69,2. 71 ± 0. 89,2. 52 ± 0. 30,5. 45±1. 41,respectively. The ratio in treatment A group was significantly higher than those in model A group(P<0. 05). There was no significant difference between treatment B and model B group ( P>0. 05 ) . Conclusion: Inguinal subcutaneous immunotherapy has a good effect on this disease. It spends short time ,has simple operation and good feasibility,which is a novel treatment method for AR in mice.

OBJECTIVE: To explore the changes of microRNAs in nasal mucosa after the specific immunotherapy (SIT) for allergic rhinitis (AR) in mice. METHOD: Female BALB/c mice, 6-8 weeks of age, were randomly divided into control group, model group and treatment group. AR model were established by intraperitoneal injection and intranasal challenge of ovalbumin and SIT was performed by inguinal subcutaneous injections. AR symptom scores were documented. The eosinophils (EOS) in the nasal mucosa were measured. Ovalbumin-specific IgE (OVA-sIgE) in the serum and expression of interferon-γ and interleukin-4 in the nasal lavage were measured by enzyme-linked immunosorbent assay meanwhile the ratio of interferon-γ and interleukin-4 was calculated. The microRNAs in the nasal mucosa were preliminary screened by microRNA gene microarray. Comparing with model group, the Fold changes of microRNA of the treatment group were ≥ 2.0 and the P < 0.05. MicroRNA target genes were predicted with GeneSpring 12.5 software. We took the intersection between genes in the signal pathway which associated with immune response,inflammation and target genes. The MEV-4-6-0 and Cytoscape_v2. 8. 2. software was applied to perform the cluster analysis and target gene regulatory networks maps. RESULT: The model of AR in mice and its SIT were successful. Comparing with the model group, the Fold changes of 15 microRNAs, of which 9 microRNAs were up-regulated and 6 microRNAs were down-regulated, were ≥ 2.0 in treatment group (P < 0.05). Cluste analysis showed clearly that microRNAs in the treatment group and model group respectively aggregated in two branches. The 15 microRNAs had 5302 target genes, of which, 451 genes were related more with SIT by the intersection. One microRNA can regulate many target genes, and one gene can also be affected by many microRNAs. Their synergistic effects may be involved in the mechanism of SIT. CONCLUSION The expressions of microRNAs are changed in nasal mucosa after SIT for AR in mice and we can speculate that microRNAs are involved in the process of SIT for AR. Bioinformatics methods can diminish the scope of target genes of microRNAs, which will help us studying the effect of changed microRNA on its relative target genes after SIT, and make us better understanding the mechanism of the disease and its SIT.
Administration, Intranasal ; Animals ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Eosinophils ; immunology ; Female ; Immunoglobulin E ; blood ; Immunotherapy ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Mice ; Mice, Inbred BALB C ; MicroRNAs ; metabolism ; Nasal Mucosa ; drug effects ; metabolism ; Ovalbumin ; Rhinitis, Allergic ; therapy

Objective:To analyze the mistakes in the application of tuina therapy in the rehabilitation of stroke,in order to elevate the clinical effects in the treatment of stroke by tuina.Methods:The issues are found out by searching and analyzing the relevant articles on tuina treatment of stroke.Results:In the tuina treatment of stroke,mistakes exist in the intervening time of the treatment,in the emphasis of tuina with negligence of rehabilitation,in the assessment of therapeutic effects,in the selection of the manual techniques and acupoints,and in the negligence of health education.Conclusion:In the tuina treatment of stroke,it is advisable to start treatment in the early stages,use the manual techniques in different phases,to cooperate with rehabilitation at the proper time and to emphasize health education,in order to avoid or reduce the occurrence of misuse syndrome,and eliminate the appearance of disuse syndrome,for displaying the unique advantage of tuina therapy fully and promoting the innovation and development of TCM rehabilitative techniques.

Objective To study the preparation method and analytical technique of hyperoside from Flos Abelmoschus manihot.Methods Hyperoside was isolated and purified by solvent extract and chromatography, whose structure was determined by 1H-NMR and 13C-NMR. The purity was analyzed by TLC and HPLC.Results The TLC showed that the hyperoside had no impurity spot. The HPLC indicated that the purity reached more than 98.5%.Conclusions The mothod of isolation and purification for hyperoside reported in this paper was simple and economical.

OBJECTIVE: To explore the effects of simultaneous multi-level surgery intervention on the changes of the oxidative stress and the cognitive function in young and middle-aged patients with moderate-severe OSAHS. METHOD: One hundred and sixteen young and middle-aged patients with moderate-severe OSAHS received simultaneous multi-level surgery. PSG,malondialdehyde (MDA), advanced oxidation protein products (AOPP) and 8-isoprostaneF2α (8-iso-PGF2α) were tested, together with the evaluation of the questionnaires of Montreal cognitive assessment (MoCA), mini-mental state examination (MMSE) and Epworth sleepiness scale (ESS) before and 6 months after surgery. RESULT: (1) The AHI, TS90% and LSaO₂ were all improved significantly 6 months after operation. No severe complications were observed in any patient. (2) The preoperative MoCA and MMSE scores were (23.52 ± 2.64) and (26.54 ± 1.07) respectively. The cognitive impairment were detected in OSAHS patients. MoCA was negatively correlated with AHI, TS90%, MDA, AOPP and 8-iso-PGF2a, while positively correlated with LSaO₂ (P < 0.05, r = -0.233, -0.190, -0.256, -0.247, -0.221 and 0.215, respectively). Cognitive impairment was mainly manifested as disorder in the visual space and execu- tive function, memory and delayed recall (mainly short-term memory). (3) MoCA and MMSE scores were both increased (P < 0.05) after the operation. MoCA showed negative correlations with MDA, AOPP, 8-iso-PGF2α, AHI and TS90%, and positive correlation with LSaO₂ (P < 0.05, r = -0.495, -0.463, -0.397, -0.332, -0.292 and 0.336, respectively). CONCLUSION Oxidative stress and cognitive dysfunction are closely related in young and middle-aged patients with Moderate-severe OSAHS. Oxidative stress may play an important role in cognitive function of young and middle-aged patients with Moderate-severe OSAHS. Multi-level surgery can improve oxidative stress so that to improve cognitive impairment.
Adult ; Cognition ; Cognition Disorders ; complications ; Humans ; Middle Aged ; Oxidative Stress ; Sleep Apnea, Obstructive ; surgery

Objective: To construct humanized anti-CD19 chimeric antigen receptor T cells and investigate its ability to kill leukemia cells in vitro and in vivo. Methods: Humanized anti-human CD19 antibody with a high affinity was obtained based on mouse anti-human CD19 antibody (FMC63). Humanized CD19 CAR-T cells (hCART19) were constructed through transfection of lentivirus carrying a CAR sequence of humanized anti-CD19 scFv into human peripheral CD3⁺ T cells. The ability of hCART19 to kill leukemia cells and secrete cytokines was detected by LDH release assay and ELISA. The in vivo tumor-killing effect of hCART19 was evaluated in a leukemia mouse model. Results: Several different humanized CD19 single-chain antibodies which were constructed by IMGT database were expressed in the eukaryotic expression vector and purified followed by acquiring humanized CD19 antibody (Clone H3L2) with similar binding ability to FMC63. Humanized CD19 CAR lentivirus vector was constructed and transfected into T cells to obtain hCART19 cells. The LDH release experiment confirmed that the killing rate of target cells was increased gradually along with the increased E/T ratio. When the ratio of E/T was 10∶1, the killing rate of target cells by hCART19 reached a maximum. When Raji cells were used as target cells, the hCART19 cells group had a significantly higher kill rate [(87.56±1.99)%] than the untransduced T cells group [(19.31±1.16)%] and the control virus transduced T cells group [(21.35±1.19)%](P<0.001). ELISA analysis showed that the secretion of IL-2 [ (10.56±0.88) pg/ml] and IFN-γ [ (199.02±12.66) pg/ml] in the hCART19 cells group were significantly higher than those in the untransduced T cells group [IL-2: (3.55±0.26) pg/ml; IFN-γ: (37.63±0.85) pg/ml] and the control virus transduced T cells group [IL-2: (2.92±0.32) pg/ml; IFN-γ: (52.07±3.33) pg/ml](P<0.001). The above experiments also showed similar results when CHO-K1-CD19 cells were used as target cells. Moreover, in a human leukemia xenograft animal model, the results showed that mice in the untransduced T cells group and the control virus transduced T cells group all died within 20 to 30 days, and the hCART19 cell group survived >40 days, which was more than the survival time of the other two groups of mice. The difference was statistically significant (χ²=11.73, P=0.008). Conclusion Humanized CD19 CAR-T cells with anti-leukemic activity have been successfully constructed, which will lay a foundation for clinical studies in the future.

Objective:To establish a disk (CD) microfluidic chip detection platform for the rapid detection of CALR-1 and CALR-2 mutations in patients with cerebral infarction, and summarize its clinical application value.Methods:Based on microfluidic technology and loop mediated isothermal amplification technology, a CD microfluidic chip detection platform for simultaneous detection of CALR-1 and CALR-2 gene mutations were established, and the sensitivity, specificity, repeatability and accuracy of the platform were verified. A total of 124 patients with cerebral infarction treated in Huashan Hospital, Shanghai Medical College, Fudan University from November 2019 to March 2021 were prospectively selected into the experimental group; and 80 healthy subjects were included in the control group. The CALR-1 and CALR-2 gene mutations in anticoagulant peripheral blood samples were detected by the CD microfluidic chip. Each chip could detect 4 samples at the same time and synchronously detect 3 indexes of each sample. The detection results could be obtained after isothermal amplification for 40 min. At the same time, sequencing method was used to verify the test results, and the consistency of the results of the two detection methods was compared.Results:Using this CD microfluidic chip platform, the synchronous amplification of 3 indexes in the sample could be completed within 40 min without the need of thermal circulation, and the whole detection process of the sample could be completed within 60 min. For samples with a high concentration of target nucleic acid, typical positive signals could be visualized after amplification for 10 min, and the test results would be available within 30 minutes after receiving the samples. The detection sensitivity of CD microfluidic chip method for CALR-1 and CALR-2 mutation load concentration was 1.0% and 0.5% respectively. Nonspecific amplification was not observed for the non-target nucleic acid samples, indicating the high specificity of this method. The coincidence rates of intra and inter batch repeatability were 100% (20/20) respectively. Two samples with CALR gene mutation were found in the cerebral infarction group, both of which were CALR-1 mutations (L367fs*46). There was no CALR-1 or CALR-2 mutation in the control group. The detection results of CD microfluidic chip method were completely consistent with the sequencing verification results (100% [204/204]).Conclusions:The CD microfluidic chip method could be used for the detection of CALR-1 and CALR-2 gene mutations in clinical samples of patients with cerebral infarction. This method has the advantages of high detection sensitivity, good detection specificity, fast detection speed and high detection flux, which is helpful to clarify the etiology of patients with cerebral infarction.

Objective:To explore the relationship between different gestational weeks of transvaginal ultrasound-guided fetal reduction and abortion in patients with multiple pregnancies after embryo transfer, and to seek the best gestational age for fetal reduction.Methods:The datas of 486 pregnant women with multiple pregnancies after embryo transfer in the Second Hospital of Hebei Medical University from January 2012 to December 2020 were retrospectively analyzed. The relationship between gestational weeks of fetal reduction and abortion rate was analyzed by curve fitting, threshold effect and multivariate logistic regression analysis.Results:After adjusting for age, infertility type, infertility years, number of births, abortion times, body mass index(BMI), various infertility and sterility factors, endometrial thickness on the day of transformation, monozygotic twins and reduction methods, when the gestational age was less than 8.43 weeks, the abortion rate increased significantly with the increase of reduction gestational age, and the abortion rate increased by 221% ( OR=3.21, 95% CI=1.47-6.99, P=0.003 3). When the gestational age of reduction ≥8.43 weeks, the abortion rate tended to be stable and did not increase ( OR=0.81, 95% CI=0.54-1.22, P=0.317 7); meanwhile, in stratified analysis, the OR value of the BMI ≥24 kg/m 2 was 12.38, and that of BMI <24kg/m 2 was 1.91, P=0.053 9. Conclusions:There is a non-linear relationship between gestational age and abortion rate of ultrasound-guided multiple pregnancy reduction in patients with embryo transfer. The abortion rate increases significantly with the increase of gestational age before 8.43 weeks of gestation. It is recommended to carry out the operation as early as possible before 8 weeks of pregnancy. The effect of BMI on the abortion rate of patients with fetal reduction needs further study.

Objective To investigate the feasibility and efficacy of endoscopic catheterization of ileus tube combined enterectomy for the treatment of elderly patients with acute sigmoid volvulus. Methods From August 2015 to August 2017,27 cases of elderly patients with acute sigmoid volvulus received treatment of endoscopic placement of ileus tube combined enterectomy in Pudong New Area Gongli Hospital.Retrospectively collected clin-ic parameters of pre-catheterization,post-catheterization,enterectomy and postoperative follow-up;the success rate of catheterization and enterectomy,compression efficiency,post-operative complications,and efficacy of the com-bined therapy were analyzed. Results Emergency catheterizations of anorectal ileus tube were succeeded in 27 (100%)patients;one patient who developed intestinal gangrene received emergency enterectomy,the hemogram and internal environmental disturbance of the 26 cases were significantly relieved,and the efficacy rate of decom-pression was 96.3%. The success rate of was enterectomy was 100%;the incidence of severe complications and mortality were 22.2% and 3.7%,respectively. During the follow-up,2(7.4%)patients developed ileus,and the efficacy rate of the combined therapy was 85.2%. Conclusions The catheterization of ileus tube could effectively depress intestinal pressure. The success rate of enterectomy was significantly improved and postoperative mortality was reduced. The therapy of endoscopic ileus tube catheterization combined enterectomy is a safe and effective method for the treatment of acute sigmoid volvulus in the elderly.