Objective To establish a stable cell line secreting human IgE Cε2-4 protein, and in-vestigate the binding capacity of receptor FcεR Ⅰ Methods The E24 gene was derived from SKO-O07 cell line, and was then cloned into pcDNA3.1 (+) (signal peptides were synthesized and fused at the 5'-end of E24 gene) or pCMV-L vector. After transient transfection into 293T cell, the secreted F24 protein was ana-lyzed by sandwich ELISA. The best vector was chosen to be transfected into CHO cells with LipofectAMI-NETM 2000 reagent. After being selected by G418 and subcloned three times by limited-dilution method, two stable cell lines were established. E24 gene was amplified by RT-PCR, and the E24 protein in the superna-tant was identified by ELISA. Besides, the binding capacity of FceR ⅠⅡ was analyzed by flow cytometry method. Results Three mammalian expression vector SP-E24-F3. 1, SP lI-E24-P3.1 and E24-PL were constructed and transient transfected to 293T cells. The output of E24 protein in the supernatant were 19.1, 19.4 and 8.7 μg/ml, respectively. Then the vector SP IX-E24-P3.1 was transfected into CHO cells. Final-ly, two single clones secreting E24 protein were stably obtained. The output of E24 were all at least 25 μg/ml. RT-PCR could detect the E24 gene from one of the two clones. Furthermore, flow cytometry results showed that E24 could bind the receptor in a dose-dependent manner. Conclusion Two stable cell line se- creting E24 protein were obtained, while E24 could specifically bind FcεR Ⅰ.

Objective To design and express a novel peptide based on ricin toxin antibody in E. coli, and to evaluate its biological activity. Methods Based on the crystal structure of ricin toxin A chain (RTA) and the RTA-rRNA interact in the complex model, the steric conformation of RTA was theoretical modeled and its functional domain was preliminarily determined. The humanized single-domain RTA antibody was designed rationally by computer-guidod molecular design method. Its coding sequence was ob- tained by overlapping extension PCR, and cloned into the pET-32a vector. The fusion protein was then ex-pressed in E. coli BL21 (DE3), identified by Western blot, and purified with Ni-NTA agarose. The binding and neutralizing activity of this novel peptide for riein was evaluated by competitive ELlSA assay and MTT assay. Results A recombinant human single-domain antibody expressing a polypeptide against RTA in the CDR3 loop was designed. The fusion protein was successfully expressed in E. coll. The purified protein can bind to ricin, and neutralize its activity in SP2/0 viability assay. Conclusion The success of the novel pep-tide based on riein toxin antibody provides a novel method to develop new generation of ricin antagonists.

Objective To analyze the cerebral autoregulation capability in patients with chronic insomnia disorder (CID).Methods Sixty CID patients (54 with generalized anxiety disorder) and 40 healthy controls were enrolled in this study.Polysomnography was done in all the participants.Noninvasive continuous cerebral blood flow velocity of bilateral middle artery and arterial blood pressure were recorded simultaneously using transcranial Doppler and a servo-controlled plethysmograph.Transfer function analysis was used to derive the autoregulatory parameters, including phase difference and coherence function.Results The phase difference values of CID patients with generalized anxiety disorder were significantly lower than that of the healthy controls ((46.89±15.39)°vs (56.00±12.05)°, t=3.439, P=0.001).In the correlation analysis, we further found that there was no correlation among phase difference values and the score of Hospital Anxiety and Depression scale.Conclusions The dynamic cerebral autoregulation was compromised in CID patients with generalized anxiety disorder regardless of the degrees of anxiety and depression.Dynamic cerebral autoregulation may be a potential therapeutic target in improving neurological symptoms in patients with CID.

Objective:To explore the factors affecting the antiviral treatment efficacy of acquired immunodeficiency syndrome (AIDS) patients.Methods:A total of 107 patients diagnosed with human immunodeficiency virus (HIV) infection in the clinic of Beihai People′s Hospital from January 2016 to June 2018 were selected.The patients were divided into two groups according to whether they voluntarily accepted traditional Chinese medicine treatment, including treatment group who received highly active anti-retroviral therapy (HAART) and traditional Chinese medicine prescription of Ping Gan Jie Du (42 cases), and control group who were only treated with HAART (65 cases). The virological and immunological responses were compared between the two groups at 48 weeks of treatment. The interleukin-28B (IL-28B) rs12979860 genotypes were measured by using the direct sequencing of polymerase chain reaction products. Logistic regression was used to analyze the influencing factors of antiviral efficacy in AIDS patients.Comparison between groups was performed by independent sample t test、matched sample t test or chi-square test. Results:At week 48 of treatment, 41 (97.62%) of the 42 patients in the HAART plus Ping Gan Jie Du group obtained virological response, while 58 (89.23%) of the 65 patients in the HAART group alone acquired virological response, which was not significantly different ( χ2=0.100, P>0.05). The numbers of CD4 + T lymphocytes increased at week 48 of treatment in the HAART plus Ping Gan Jie Du group and HAART group were (244.32±101.83)/μL and (211.56±112.50)/μL, respectively. The was no statistically significant difference ( t=1.522, P>0.05). Among the 92 patients with IL-28B CC genotype, 88 (95.65%) acquired virological response, while 11 of the 15 patients with non-IL-28B CC genotype acquired virological response, which was not significantly different ( χ2=0.394, P>0.05). And CD4 + T lymphocytes in patients with IL-28B CC genotype increased ((229.72±101.17)/μL), which was higher than that without IL-28B CC genotype ((173.40±89.64)/μL), with statistically significant difference ( t=2.028, P=0.045). Multivariate logistic regression analysis showed that baseline CD4 + T lymphocyte count≤200/μL, IL-28B CC genotype, and treatment plan including protease inhibitor were helpful to improve the antiviral efficacy. Conclusion:Baseline CD4 + T lymphocyte count ≤200/μL, IL-28B CC genotype, and protease inhibitor in HAART regimen are the influencing factors of antiviral efficacy in AIDS patients.

Objective:To evaluate the effect of electroacupuncture on the heme oxygenase-1 (HO-1)/PTEN-induced putative kinase 1 (PINK1)/Parkin signaling pathway during acute kidney injury in endotoxemic rats.Methods:Twenty-four SPF healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 180-220 g, were divided into 4 groups ( n=6 each) by a random number table method: control group(group C), endotoxemia group(group E), acupoint electroacupuncture+ endotoxemia group(group EE), and non-acupoint electroacupuncture+ endotoxemia group(group NE). The endotoxemia model was developed by intraperitoneal injection of lipopolysaccharide 10 mg/kg. The equal volume of normal salinewas injected in group C. LPS 10 mg/kg was intraperitoneally injected in group E. In group EE, 30-min electroacupuncture was performed at bilateral Zusanli and Shenshu acupoints using disperse-dense waves with a frequency of 2/15 Hz to induce slight muscle tremor once a day starting from 5 days before developing the model, and the needle was retained until 6 h after injection. Electroacupuncture was performed at the points 0.5 cm lateral to the acupoints of Zusanli and Shenshu in group NE. The rats were anesthetized at 6 h after lipopolysaccharide injection, and blood samples from the femoral vein were obtained for determination of the serum creatinine (Cr) and urea nitrogen (BUN) concentrations(with a biochemical analyzer) and concentrations of neutrophil gelatinase-associated lipid transport protein (NGAL), interleukin-6 (IL-6), tumor necrosis factor (TNF-α) and kidney injury molecule-1(KIM-1) in serum (by enzyme-linked immunosorbent assay). Then the rats were sacrificed and kidney tissues were taken for determination of histological score of kidneys (HSK, using HE staining) and expression of HO-1, PINK1, Parkin, mitochondrial fusion protein 2(Mfn2), optic atrophy protein 1(OPA1) and mitochondrial dynamic-related protein 1 (Drp1) (by Western blot). Results:Compared with group C, serum concentrations of Cr, BUN, KIM-1, NGAL, IL-6 and TNF-α and HSK score of renal tissues were significantly increased, the expression of HO-1, PINK1, Parkin and Drp1 was up-regulated, and the expression of Mfn2 and OPA1 was down-regulated in E, EE and NE groups ( P<0.05). Compared with group E, serum concentrations of Cr, BUN, KIM-1, NGAL, IL-6 and TNF-α and HSK score of renal tissues were significantly decreased, and the expression of HO-1, PINK1, Parkin, Mfn2 and OPA1 was up-regulated, and Drp1 expression was down-regulated in group EE( P<0.05), and no significant change was found in the parameters mentioned above in group NE ( P>0.05). Conclusions:The mechanism by which electroacupuncture alleviates acute kidney injury is associated with activation of HO-1/PINK1/Parkin signaling pathway in endotoxemic rats.

Objective:To investigate the safety and efficacy of remimazolam combined with afentanyl for fiberoptic bronchoscopy.Methods:Sixty patients admitted to Chifeng Hospital for fiberbronchoscopy from January to April 2022 were selected and divided into two groups by random number table method: remimazolam group (group R) and propofol group (group P), 30cases in each group. After intravenous injection of alfentanil for anesthesia induction, group R was sedated by intravenous injection of remidazolam besylate, and group P was sedated by intravenous injection of propofol emulsion. When sufficient sedation was achieved, fiberoptic bronchoscopy was performed. The patients were scored with the Mini-Mental State Examination (MMSE) before examination and before leaving the room. The recovery rate of sedation and the recovery rate of drugs during operation were compared. Blood pressure, heart rate (HR), bispectral index (BIS), SpO 2 value and Modified Observer′s Assessment of Alertness/Sedation (MOAA/S) score were compared before induction (T 0), at the beginning of examination (T 1), immediately when fiber bronchoscope reached juga (T 2), at the end of surgery (T 3), immediately, when patients regained consciousness (T 4). Drug onset and recovery time (time out of hospital) as well as the incidence of intraoperative and postoperative adverse reactions were recorded in both groups. Results:There was no statistically significant difference in general condition, MMSE score and examination time between the two groups (all P>0.05). There was no statistically significant difference between the two groups in the success rate of sedation and the number of sedative remedy times (all P>0.05). The number of additional drugs in group R was significantly higher than that in group P ( P<0.05). The systolic blood pressure, diastolic blood pressure and BIS values of patients in group P at T 1 and T 2 were significantly lower than those in group R (all P<0.05). After administration, the MOAA/S score of the two groups began to decrease, and the decrease of the P group was significantly greater than that of the R group, and the MOAA/S value of the patients was the lowest at the 3rd and 4th minutes after administration, respectively. The time from the beginning of administration to the MOAA/S score ≤3 in group P was significantly shorter than that in group R (all P<0.05). The incidence of pain and respiratory depression after injection in group P was significantly higher than that in group R ( P<0.05). Conclusions:The application of afentanil combined with remimazolam in the patients undergoing fiberoptic bronchoscopy has good sedative effect and high anesthesia quality, and has no obvious effect on cognitive function and few adverse reactions, so it is safe and effective.

OBJECTIVE: To discuss a effective method of nasal septum deviation. METHOD: One hundred and sixteen subjects with nasal septum deviation were divided into treatment group (69 subjects) and control group (47 subjects) randomly. The combination therapy of correction of deviated nasal septum and plasma radiofrequency ablation were used in the treatment group. The combination therapy of deviated nasal septum and partial inferior turbinectomy were used in the control group. The data were analyzed by statistical method. RESULT: The effective rate of physical signs and symptoms of the treatment group was 100.0%, while the control group was 85.1%. There was significant difference of two effective rates (P < 0.05). Furthermore, hyperventilation and nasal adhesion were not happened in treatment group 6 months after treatments. CONCLUSION The correction of deviated nasal septum and plasma radiofrequency ablation combination therapy had the satisfied and safety treatment effect, which was and plasma radiofrequency ablation combination therapy had the satisfied and safety treatment effect, which was easy for observation as well as the minimal tissue damage. The combination therapy method was according with the principle of functional minimally invasive surgery.
Adult ; Catheter Ablation ; Combined Modality Therapy ; Female ; Humans ; Male ; Middle Aged ; Nasal Septum ; abnormalities ; surgery ; Reconstructive Surgical Procedures ; Young Adult

Objective:To evaluate the effect of electroacupuncture (EA) on pyroptosis in renal tubular epithelial cells of rats with acute kidney injury (AKI) induced by endotoxin.Methods:Twenty-four healthy clean-grade Sprague-Dawley rats of either gender, aged 6-8 weeks, weighing 160-182 g, were divided into 4 groups ( n=6 each) using a random number table method: control group (group C), group AKI, EA plus AKI group (group EA), sham EA at non-acupoint plus AKI group (group SEA). The model of endotoxemia was established by intraperitoneally injecting 10 mg/kg lipopolysaccharide.Bilateral 30 min EA stimulation of Zusanli and Shenyu (according to atlas of animal acupoint) was performed starting from 5 days before establishing the model (once a day) and at 30 min before lipopolysaccharide administration on the day of establishing the model, with disperse-dense waves, frequency of 15 Hz, and the needle was kept until 6 h after injection of LPS in group EA.EA was performed at the points 0.5 cm lateral to the acupoints of Zusanli and Shenyu in group SEA.At 6 h after LPS injection, blood was taken from the heart, and the concentrations of serum blood urea nitrogen (BUN) and creatinine (Cr) were detected by an automatic biochemical analyzer, and the serum concentrations of neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) by enzyme-linked immunosorbent assay.The rats were then sacrificed, and the left renal cortex was obtained for determination of pyroptosis rate of renal tubular epithelial cells (by TUNEL). The right renal cortex was obtained to detect the expression of caspase-1 and IL-1β by Western blot, and the expression of caspase-1 mRNA and IL-1β mRNA was detected by real-time polymerase chain reaction. Results:Compared with group C, the concentrations of BUN, Cr, NGAL, KIM-1, TNF-α, and IL-6 were significantly increased, the pyroptosis rate of renal tubular epithelial cells was increased, the expression of caspase-1 and IL-1β protein and mRNA in the renal cortex was up-regulated in group AKI ( P<0.05). Compared with group AKI, the concentrations of BUN, Cr, NGAL, KIM-1, TNF-α, and IL-6 were significantly decreased, the pyroptosis rate of renal tubular epithelial cells was decreased, the expression of caspase-1 and IL-1β protein and mRNA in the renal cortex was down-regulated in group SEA ( P>0.05). Conclusion:The mechanism by which EA reduces AKI may be related to inhibiting pyroptosis in renal tubular epithelial cells of rats.

Objective To analyze the spatial distribution of HIV/AIDS epidemics among injecting drug users (IDUs) in China from 2005 to 2011 and to understand its changing trend.Methods Using data on people living with HIV and AIDS through injecting drug between 2005 and 2011 to analyze the demographic characteristics of injecting drug users.Analysis on spatial correlation (provincial level and country level) and median center of hot spots (country level) were conducted by Arcgis software.Results Sex ratio (male/female) and registered place ratio (province/other provinces) reduced as time went by,with the ratios in 2011 as 6.75 and 7.01 respectively.Tape ratio of the disease between HIV and AIDS showed an upward trend (Z=26.880,P＜0.01).Since 2005,the identified numbers of HIV/AIDS and the spatial correlation and hot spots in provincial level had reduced,the numbers of provincial hot spots were two from 2009 to 2011 (Sichuan and Yunnan provinces) at the national level.However,the spatial correlation and hot spots at the provincial level had an increasing trend.Between 2005 and 2011,the Western Median Centers of hot spots was located in Xinjiang province while the Southwestern Median Center of hot spots tended to move towards the north.Conclusion The demographics changes of HIV/AIDS infection among injecting drug users seemed to be regular from 2005 to 2011.Spatial correlation at the provincial level was reducing.However,the spatial correlation and the numbers of hot spots at the country level increased,with hot spots at the country level tended to move from the border areas towards inland.

PURPOSE: The long non-coding RNA H19, a conservatively imprinted gene, acts as a molecular sponge for the let-7 family, which has been identified as a set of tumor suppressors. However, the combined prognostic value of H19 and let-7a signature in breast cancer patients remains unclear. METHODS: In this research we assessed the prognostic value of the combined H19 and let-7a signature in breast cancer patients by retrospectively reviewing that data of 79 patients who underwent neoadjuvant chemotherapy; we also investigated the expression and function of H19 in breast cancer cell lines in vitro. Survival data were calculated using the Kaplan-Meier method and compared using the log-rank test. Univariate and multivariate survival analyses were conducted using the Cox proportional hazards regression method. As determined using X-tile, the optimal cutoff value for the risk score to assess progression-free survival (PFS) based on the combined signature was –0.1. RESULTS: Patients with an overall positive treatment response had higher let-7a and lower H19 levels. In addition, let-7a expression was negatively correlated with H19 expression. Patients with a risk score of >–0.1 had shorter overall survival and PFS. In vitro data showed that chemoresistant cell lines exhibit higher H19 and lower let-7a levels and knockdown H19 restores paclitaxel sensitivity. CONCLUSION: Our results suggest that the combined let-7a and H19 signature is a novel prognostic factor for breast cancer patients treated with neoadjuvant chemotherapy.
Breast Neoplasms* ; Breast* ; Cell Line ; Disease-Free Survival* ; Drug Therapy ; Humans ; In Vitro Techniques ; Methods ; Neoadjuvant Therapy ; Paclitaxel ; Porifera ; Prognosis ; Retrospective Studies ; RNA, Long Noncoding