Objective To construct a stable synovial cell line MH7A from rheumatoid arthritis(RA)patients using lentiviral vectors that interfere with the expression of tumor necrosis factor receptor associated factor 2(TRAF2),and to study the role of TNF-α-TRAF2 signaling in MH7A abnormal proliferation.Methods Based on the design principles of human TRAF2 gene sequence and shRNA sequence,three pairs of TRAF2 shRNA interference se-quences were designed and synthesized.The primers were annealed by PCR,and a linear vector was obtained by double enzyme digestion PLKO.1-puro.The linearized vector was connected to the annealed primers through Solu-tion I,and the connected products were introduced into receptive cells.The plates were coated,and positive colo-nies were selected for sequencing.Three different recombinant plasmids of PLKO.1-TRAF2-shRNA lentivirus were constructed,and lentivirus packaging plasmids was used to package logarithmic growth phase HEK 293T cells.Vi-rus solution was collected to infect MH7A cells.At the same time,puromycin was used to screen MH7A stable transgenic strains with low TRAF2 expression.CCK-8 method,Western blot,and qPCR were used to detect the proliferation function of MH7A induced by TNF-α and low expression of TRAF2,as well as downstream signal TRAF2,P65 protein expression and mRNA levels.Results PLKO.1-TRAF2-shRNA(1),PLKO.1-TRAF2-shR-NA(2),and PLKO.1-TRAF2-shRNA(3)lentivirus vector plasmids and control group lentivirus vector plasmids PLKO.1-puro were successfully constructed.The three TRAF2-shRNA lentivirus vector plasmids and control group lentivirus vector plasmids PLKO.1-puro were respectively introduced into the lentivirus packaging plasmid of HEK 293T to obtain virus solution.After infecting MH7A cells with the virus solution,they were treated with puromycin(2.00 μ G/mL)screening and obtaining MH7A stable transgenic plants after 2 days.Through qPCR and Western blot results,it was found that the expression of TRAF2 mRNA and protein in PLKO.1-TRAF2-shRNA(1)MH7A stably transfected cells was significantly reduced compared to the negative control group.The results of CCK-8 and Western blot showed that after knocking down TRAF2 in MH7A,the proliferation of MH7A cells with low TRAF2 expression induced by TNF-α and the phosphorylation level of P65 were significantly reduced.Conclusion A sta-ble transgenic strain of PLKO.1-TRAF2-shRNA(1)MH7A cells was successfully constructed to investigate the role of TNF-α-TRAF2 signal activation in mediating abnormal proliferation of RA synovial cells.

In combined orthodontic-orthognathic treatment, the maxillary palatine suture is closed in most patients with insufficient maxillary width, and bony expansion of the maxilla cannot be achieved by dental expansion or rapid palatal expansion (RPE) which causes buccal inclination of the maxillary posterior teeth leading to unstable results. Therefore, segmental LeFort Ⅰ osteotomy and surgically assisted RPE are often used in clinical practice. In recent years, with the application of implant anchorage technology, implant anchorage assisted RPE has been gradually applied in orthognathic treatment. This article reviewed the indications, contraindications, complications, efficacy and long-term stability in different treatment approaches including segmental LeFort Ⅰ osteotomy, surgically assisted RPE and implant-supported maxillary skeletal expansion.

The nomenclature of congenital auricular malformations is complex and confusing, which brings great inconvenience to clinical work and scientific research. This study comprehensively summarized and sorted out the existing Chinese and English names of congenital auricular malformations, providing reference basis for the scientific classification and clinical treatment of congenital auricular malformations. Sixty-seven papers on the topic of congenital auricle malformations were collected from databases such as CNKI, Wanfang Database, VIP Database, PubMed, Web of Science, and ScienceDirect by using the PRISMA method. The two core issues of morphological and anatomical naming and classification of congenital auricle malformations were sorted and summarized based on these articles. In this study, the existing Chinese and English names of congenital auricular malformations were summarized, and a preliminary classification was made at the morphological and anatomical levels, and the embryological origin of the external auricle and its substructures was elaborated. Based on the result of this study, a comprehensive analysis can be conducted on the malformations of the ear structures and their embryological origins in order to explore the etiology of congenital ear malformations. It is expected that scientific and standardized naming and classification method can be developed, and reasonable and effective standardized treatment can be formulated for congenital auricular malformations based on this study.

The nomenclature of congenital auricular malformations is complex and confusing, which brings great inconvenience to clinical work and scientific research. This study comprehensively summarized and sorted out the existing Chinese and English names of congenital auricular malformations, providing reference basis for the scientific classification and clinical treatment of congenital auricular malformations. Sixty-seven papers on the topic of congenital auricle malformations were collected from databases such as CNKI, Wanfang Database, VIP Database, PubMed, Web of Science, and ScienceDirect by using the PRISMA method. The two core issues of morphological and anatomical naming and classification of congenital auricle malformations were sorted and summarized based on these articles. In this study, the existing Chinese and English names of congenital auricular malformations were summarized, and a preliminary classification was made at the morphological and anatomical levels, and the embryological origin of the external auricle and its substructures was elaborated. Based on the result of this study, a comprehensive analysis can be conducted on the malformations of the ear structures and their embryological origins in order to explore the etiology of congenital ear malformations. It is expected that scientific and standardized naming and classification method can be developed, and reasonable and effective standardized treatment can be formulated for congenital auricular malformations based on this study.

Objective To investigate serum levels of soluble apoptosis-related factor ligand(sFasL)and stromal cell derived factor 1(SDF-1)in patients with aplastic anemia(AA)and their correlation with immunosuppressive treatment.Methods Forty-three AA patients who received immunosuppressive therapy for half a year were selected as the observation group,and another 43 healthy subjects at the same period were selected as the control group.Patients in the observation group received immunosuppressive therapy and hematopoietic propoietic therapy,and patients were divided into the ineffective group and the effective group according to the efficacy.Clinical data of patients were collected,and serum levels of sFasL,SDF-1,tumor necrosis factor-α(TNF-α)and interleukin-8(IL-8)were detected by enzyme-linked immunosorbent assay.The correlation between serum sFasL and SDF-1 levels was analyzed.Multivariate Logistic regression analysis was conducted to analyze influencing factors of immunosuppressive treatment in AA patients.The predictive values of serum sFasL and SDF-1 on immunosuppressive treatment were analyzed by receiver operating characteristic(ROC)curve.Results Serum levels of sFasL and SDF-1 were higher in the observation group than those in the control group(P＜0.05).Serum levels of TNF-α,IL-8,sFasL and SDF-1 were higher in the ineffective group than those in the effective group(P＜0.05).Serum sFasL level was positively correlated with SDF-1 level in AA patients(r=0.534,P＜0.001).Increased serum sFasL and SDF-1 levels were independent risk factors for ineffective immunosuppressive therapy in AA patients(P＜0.05).The area under the curve(AUC)of serum sFasL and SDF-1 combined to predict the immunosuppressive treatment effect of AA patients was 0.973(95%CI:0.872-0.999),which was better than that of single diagnosis(P＜0.05),and its sensitivity and specificity were 94.44%and 96.00%,respectively.Conclusion Serum levels of sFasL and SDF-1 are significantly increased in patients with AA.The combined detection of sFasL and SDF-1 has high predictive value for the immunosuppressive treatment effect of AA.

Objective:To understand the effect of vitamin B1 on lymphocyte function in simulated microgravity.Methods:Splenocytes of mice were isolated,and the rotatary cell culture system was used to simulate microgravity.Lymphocytes were stimulated with mitotic agents Concanavalin A,and cells were treated with different concentrations of vitamin B1,proliferation indexes of lympho-cytes and levels of cytokines in supernatant were detected.Results:Simulated microgravity could inhibit proliferation of splenic lym-phocytes,and decrease levels of cytokines,while vitamin B1 could promote lymphocyte proliferation and cytokines production in cells cultured in simulated microgravity in a dose dependent manner.Conclusion:Vitamin B1 may attenuate the inhibitory effect of simulated microgravity on lymphocytes by regulating cell proliferation and secretion of cytokines.

Objective This study aims to predict the coronavirus disease 2019(COVID-19)epidemic in Xi'an based on SEAIQR model and Dropout-LSTM model,and to provide a scientific basis for evaluating the effectiveness of the"dynamic zero-COVID policy".Methods Considering a large number of asymptomatic infections,the changing parameters,and control procedures,we developed a time-dependent susceptible-exposed-asymptomatic-infected-quarantined-removed(SEAIQR)model with stage-specific interventions.Considering the time-series characteristics of COVID-19 data and the nonlinear relationship between them,we constructed a deep learning Dropout-LSTM model.The data of newly confirmed cases in Xi'an from December 9th,2021 to January 31st,2022 were used to fit the model,and the data from February 1st,2022 to February 7th,2022 were used to evaluate the model performance of forecasting.We then calculated the effective reproduction number(Rt)and analyzed the sensitivity of the different measurement scenarios.Results The peak of newly confirmed cases predicted by the SEAIQR model would appear on December 26th,2021,with 176 cases,and the"dynamic zero-COVID policy"may be achieved in January 24th,2022,with R2=0.849.The Dropout-LSTM model can reflect the time-series and nonlinear characteristics of the data,and the predicted newly confirmed cases were highly consistent with the actual situation,with R2=0.937.The MAE and RMSE of the Dropout-LSTM model were lower than those of the SEAIQR model,indicating that the predicted results were more ideal.At the beginning of the outbreak,R0 was 5.63.Since the implementation of comprehensive control,Rt has shown a gradual downward trend,dropping to below 1.0 on December 27th,2021.With the reduction of effective contact rate,the early implementation of control measures and the improvement of immunity threshold,the peak of newly confirmed cases will continue to decrease.Conclusion The proposed Dropout-LSTM model forecasts the epidemic well,which can provide a reference for decision-making of the"dynamic zero-COVID policy."

Objective:To investigate the clinical infection status and trends in drug resistance of a rare pathogen Kluyveromyces marxianus ( Candida kefyr), and to provide experience for clinical diagnosis and treatment. Methods:Morphological and molecular biological identification tests and in vitro microdilution drug susceptibility test were conducted on a Kluyveromyces marxianus strain recently isolated from the midstream urine sample of a patient with urinary calculus in the Fungal Laboratory, Changhai Hospital. The ultrastructural damage of the strain caused by different antifungal drugs was observed by scanning electron microscopy. A retrospective analysis was conducted on clinical cases of Kluyveromyces marxianus infection in Changhai Hospital from 2009 to 2021. Results:The isolated strain formed smooth, soft, cheese-like yeast colonies on the Sabouraud′s agar medium, and ovoid or slender spores were observed under the microscope. Morphological analysis, mass spectrometry and sequencing analysis identified the strain as Kluyveromyces marxianus. The drug susceptibility test showed that minimum inhibitory concentrations (MICs) of amphotericin B, fluconazole, itraconazole, voriconazole, posaconazole, flucytosine, caspofungin, and micafungin were 0.5, 0.5, 0.03, ≤ 0.03, 0.06, 0.5, ≤ 0.016, and 0.06 μg/ml, respectively. Under the scanning electron microscope, the strain was ovoid to slender before antifungal drug treatment, with a size of (3.0 - 6.5) μm × (5.5 - 11.0) μm; after 24-hour treatment with antifungal drugs at the dose of 1 μg/ml, cell membrane shrinkage was more obvious under the treatment with posaconazole, which exhibited a stronger destructive effect on the strain compared with amphotericin B and voriconazole. From 2009 to 2021, 8 cases of Kluyveromyces marxianus infection were collected, including 6 males and 2 females; the Kluyveromyces marxianus strains were isolated from ascites in 3 cases, bronchoalveolar lavage fluid in 1 case, sputum in 2 cases, and midstream urine samples in 2 cases. Conclusion:For suspected Kluyveromyces marxianus infection, it is crucial to determine the pathogenic species as early as possible using various identification methods, and to collect strain as well as evaluate drug susceptibility, which will be beneficial for targeted clinical treatment.

Objective:To evaluate and analyze the health status of the elderly from physical health, mental health and social health, so as to understand the health status and health service needs of rural elderly.Methods:A total of 263 elderly people aged 60 years and over(mean aged 70.9±7.9 years, 113 males and 150 females, 125 aged 60-69 years, 98 aged 70-79 years and 40 aged 80 years and over)in Nanwangkong Village, Shaozhuang Town, Qingzhou City, Weifang City, Shandong Province were selected by cluster sampling method.The physical, mental, social and overall health status of the elderly were comprehensively evaluated and analyzed by self-designed questionnaire.Results:The overall health rate was 30%(79 cases), and the physical health, mental health and social health rates were 73.8%(194 cases), 84.0%(221 cases)and 34.6%(91 cases), respectively, in rural elderly in this area.The physical health was better in males than in females in 80~ years old groups( χ2=5.736, P<0.05). The overall health was better in males than in females in the total age group and the 60~69 years old groups( χ2=7.468 and 11.116, both P<0.01). The proportions of unhealthy, basic healthy and healthy people in the overall and the dimensions of physical health, mental health and social health had significant differences in the 60~69, 70~79 and 80~ years old groups( χ2=40.590, 29.342, 18.503 and 27.615, all P<0.01), and the Chi-square test for trend showed that there was a statistically significant downward trend of overall health grade distribution with age( χ2=21.994, 12.831, 16.570 and 22.595, both P<0.01). Conclusions:In this study, 30.0% of the rural elderly were considered healthy, 48.3% were basically healthy, and 21.7% were unhealthy.The health status of the elderly gradually deteriorates with age.The government should strengthen the multidimensional health assessment of the rural elderly and provide the comprehensive health guidance services and targeted interventions for the elderly in terms of disease control, psychological counseling and social participation.

In order to evaluate the efficacy of peroral endoscopic myotomy (POEM) for achalasia, data of 63 patients with achalasia who were treated with POEM in China-Japan Union Hospital of Jilin University from 2017 to 2021 were collected. Postoperative Eckardt score, high-resolution manometry and upper gastrointestinal radiography were compared with preoperative data. The mean age of the 63 patients was 49.0 years, and there were 31 famales. The preoperative Eckardt score was 9 (3), and the postoperative Eckardt score was 2 (2), with significant difference ( V=1 953, P<0.001). The lower esophageal sphincter pressure decreased significantly after operation compared with that before operation [9.90 (3.35) mmHg VS 26.80（13.85）mmHg， V=2 016, P<0.001]. Fifty-three patients (84.1%) had satisfactory curative effects. The incidence of adverse events was 3.2% (2/63). POEM is safe, effective and minimally invasive for the treatment of achalasia.