Objective To summarize the experience of isolating influenza virus from MDCK cell , and to optimize the isolation method for influenza isolation and raise the ability of influenza surveillance. Methods Sixty-one cases of influenza viruses positive specimens identified with PCR method by the influenza network laboratory in Xianyang CDC were used to infect the MDCK cells. The influenza viruses-infected MDCK cells were collected and characterized by hemagglutination test and hemagglutination inhibition assay. Meanwhile , different concentrations of glutamine liquid were added to the medium to investigate the effect of glutamine on the growth of MDCK cells. Results Twenty five flu strains were isolated from 61 cases influenza viruses positive specimens , of which 18 were A (H3N2) subtype strains and 7 were A (H1N1) subtype strains. The titer of HA was higher than 1∶16, and the titer of HI was higher than 1 ∶ 640. The proliferation rate of MDCK cells was low, and the cell activity was also decreased at 48 h after incubation with glutamine concentration lower than 0.1%. Conclusions The influenza viruses were isolated from the MDCK cells. The glutamine concentration is strongly associated with the activity of MDCK cells and cell apoptosis.

n of IL-6 and TNF-alpha,alleviate the inflammation of ANP.

Objective: To understand the morbidity and epidemiological characteristics of hand, foot and mouth disease (HFMD) in Xianyang in order to provide evidence for making prevention strategies. Methods: The incidence data of HFMD from 2013 to 2016 were collected from the National Disease Information Management System. The HFMD specimens were tested by polymerase chain reaction (PCR) and the results were analyzed by the Statistical Product and Service Solutions (SPSS19.0). Results: A total of 29 662 HFMD cases were reported in Xianyang from 2013 to 2016, and the average annual incidence rate was 143.62/100 000, including 406 severe cases and 4 deaths. There were statistically significant differences in incidence rate among the four years (P<0.01). The peak of the incidence occurred in April to July, and the second peak occurred in October to November. The top three counties which had high incidence were Wugong, Xunyi and Jingyang. There were statistically significant differences in different counties (P<0.01). The age group of 0-3 year scattered children has the most reported cases, a total of 26 399 cases, accounting for 89.00%. The number of male cases was higher than that of female cases and there were statistically significant differences between male and female (P<0.01). A total of 2 439 samples of HFMD were detected in the laboratory, the positive rate was 47.23%, among which coxsackievirus type 6 (CV-A6), coxsackievirus type 16 (CV-A16) and enterovirus A71 (EV-A71) were 38.02%, 29.91% and 21.01%. Conclusions The HFMD morbidity had obvious seasonal trend. Regional and demographic characteristics of morbidity were significant in Xianyang. The dominant pathogen was different from 2013 to 2016. The EV-A71 is easy to induce the epidemic of the HFMD.

Objective To compare the function recovery of multiple injuries combined with floating knee joint injury and simple knee joint injury,and to analyze the risk factors.Methods A retrospective case control study was conducted to analyze the clinical data of 41 patients with multiple injuries combined with Blake and McBryde Ⅱ A floating knee injury admitted to Xijing Hospital of Air Force Medical University from June 2011 to June 2017.There were 26 males and 15 females,aged 18-76 years,with an average of 34.5 years.There were 25 patients with simple knee joint injury and and 16 patients with multi-joint combined injury involving knee joint and ipsilateral hip joint or ankle joint injury.Surgical fixation was performed in different parts by external fixation,intramedullary nail and plate screw fixation.According to the Kalstr(o)m and Olerud functional evaluation criteria,the excellent and good rate of postoperative functional recovery was compared between the two groups.The surgical fixation methods of the two groups were compared.Logistic regression analysis was performed on the influencing factors of functional recovery.Results The patients were followed up for 1-7 years,with an average of 3 years.The excellent and good rate of overall functional recovery in the two groups was 68％,and the rate was 84％ in simple knee injury group and 44％ in multi-joint combined injury group (P ＜ 0.01).There was no significant difference in the ratio of intramedullary nail and plate screw fixation between the two groups (P ＞ 0.05),while the proportion of the external fixation in the multi-joint combined injury group [31％ (5/16)] was significantly higher than that in the simple knee joint injury group [16％ (4/25)] (P ＜ 0.05).Logistic regression analysis showed that the rate of external fixation was an independent factor affecting the postoperative function (OR =0.15,P ＜ 0.01).Conclusions The postoperative function in multi-joint injury patients is poorer than in the single joint injury patients.The higher rate of using external fixation in multi-joint injury patients is a risk factor.For Blake and McBryde Ⅱ A floating knee injury combined with multi-joint injury,less external fixation should be used,so as to improve the postoperative function.

Objective:To evaluate and compare the clinical efficacy and safety of three different medicated eye patches in the treatment of Demodex blepharitis. Methods:A multicenter, randomized, double-blind, parallel-controlled clinical trial was conducted.A total of 140 patients (280 eyes) with Demodex blepharitis were recruited in Shanghai Jing'an District Shibei Hospital, Xi'an Fourth Hospital and Kunming First People's Hospital from July 2021 to December 2022.The affected eyes were randomly divided into tea tree oil group, okra oil group, basal fluid control group and metronidazole group by the random number table method.Eye patches containing 20% tea tree oil, 1% okra oil, prepared base solution and 2% metronidazole were applied to the eyes for 28 days by the double-blind method.The count of Demodex was evaluated before treatment and on days 14 and 28 of treatment.Ocular surface symptoms were scored according to Ocular Surface Disease Index (OSDI). The degree of congestion at the eyelid margin and cylindrical dandruff at the root of eyelashes were scored under a slit lamp microscope.The effective rate was calculated according to the comprehensive scores above, and the adverse reactions of the subjects were observed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Shanghai Jing'an District Shibei Hospital (No.YL-20200320-05). All the subjects were informed of the significance, purpose and method of the study.Written informed consent was obtained from each subject before any medical examination. Results:All subjects completed the treatment and follow-up, and the loss to follow-up rate was 0%.After 14 and 28 days of treatment, the Demodex count was significantly decreased in all groups compared with before treatment (all at P<0.05). After 28 days of treatment, the number of Demodex in tea tree oil group, okra oil group and metronidazole group were significantly lower than that in basal fluid control group, with statistically significant differences (all at P<0.05). The OSDI score, palpebral margin congestion score and cylindrical dandruff score on 14 and 28 days after treatment in tea tree oil group, okra oil group and metronidazole group were significantly lower than before treatment, showing statistically significant differences (all at P<0.05). After 28 days of treatment, the effective rates of tea tree oil group, okra oil group and metronidazole group were 71.4%, 71.4% and 62.9%, respectively, which were significantly higher than 25.7% in basal solution control group.No serious local or systemic adverse reactions were found during the treatment and follow-up. Conclusions:Eye patches containing tea tree oil, okra oil and metronidazole have significant effects on the treatment of Demodex blepharitis, which can improve the biological environment of the palpebral margin and eliminate the inflammation related to blepharitis.

Objective:To identify and characterize one Spiroplasma strain (designated as DGKH1) isolated from the blood of a patient with sepsis. Methods:The traditional bacterial culture, staining, morphological observation, physiological and biochemical identification, 16S rRNA gene sequencing, phylogenetic analysis, genome sequencing, and the genome-related index analysis were performed to accurately determine the taxonomic status of the strain DGKH1. Antibiotic susceptibility testing was performed using a specific kit for culturing and testing Ureaplasma urealyticum/ Metamycoplasma hominis. Results:The strain DGKH1 could weakly grow on Columbia blood agar, chocolate agar, and Haemophilus chocolate 2 agar. However, it did not grow in liquid culture medium containing tetracycline (4 μg/ml), doxycycline (1 μg/ml), minocycline (1 μg/ml), josamycin (2 μg/ml), roxithromycin (1 μg/ml), clarithromycin (1 μg/ml), or telithromycin (1 μg/ml). DGKH1 resembling Metamycoplasma hominis formed "fried egg-like colonies" on Mycoplasma solid culture medium. DGKH1 could not be stained by Gram staining. When observed under transmission electron microscopy (TEM) using phosphate buffer as the matrix, the bacteria were spiral-shaped. Results of 16S rRNA gene sequence alignment showed that DGKH1 was highly similar (99.85%) to Spiroplasma eriocheiris CCTCC M 207170 T. However, the urea decomposition test was positive, which was different from all of the known Spiroplasma species. The phylogenetic analysis based on whole genome showed that DGKH1 was clustered in a small branch along with Spiroplasma eriocheiris CCTCC M 207170 T. However, the average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between the two strains were 94.14% and 56.00%, respectively, both below the threshold for prokaryotic species identification. Conclusions:DGKH1 represented a potential new species of genus Spiroplasma, closely related to Spiroplasma eriocheiris. Some microbiological characteristics of DGKH1 were similar to Mycoplasmas. However, the natural host and epidemiological data of DGKH1 need to be further studied.

Objective To explore the current status,hotspots and trends of research on the TCM regulation of cellular autophagy;To provide reference for related research.Methods The literature related to the TCM regulation of cellular autophagy was retrieved from CNKI,Wanfang Data,VIP and CBM from the establishment of the databases to December 31,2022.Authors and institutional collaboration networks were drawn using CiteSpace 5.7.R5.keywords emergence analysis and timeline view were drawn.VOSviewer 1.6.18 software was used to draw keyword co-occurrence mapping.Results A total of 2 001 articles were included,with a general upward trend in the number of publications.The cooperation teams were mainly represented by Jia Lianqun,Yang Guanlin,Song Nan and others.Research institutions with more publications included Hunan University of Chinese Medicine,Liaoning University of Chinese Medicine,and Guangzhou University of Chinese Medicine.The keywords of the literature formed 9 meaningful clusters,showing that the research hotspots in this field mainly focused on apoptosis,mitochondrial autophagy,experimental studies,signaling pathways,and herbal monomers,etc.TCM interventions that attracted much attention included electroacupuncture,moxibustion,resveratrol,curcumin,etc,mainly involving PI3K/Akt,AMPK/mTOR and other signaling pathways.Conclusion The research on cellular autophagy in the field of TCM has been highly popular in recent years.Experimental studies,molecular mechanisms,related diseases,and TCM intervention are the research hotspots in this field.The research trend is dominated by TCM monomer,TCM theory research and other directions,mainly focusing on apoptosis,oxidative stress and other pathways to carry out extensive research.

Objective To describe the significance of the pretreatment inflammatory indicators in predicting the prognosis of patients with esophageal squamous cell carcinoma (ESCC) after undergoing radical radiotherapy. Methods The data of 246 ESCC patients who underwent radical radiotherapy were retrospectively collected. Receiver operating characteristic (ROC) curves were drawn to determine the optimal cutoff values for platelet-lymphocyte ratio (PLR), neutrophil-lymphocyte ratio (NLR), and systemic immune-inflammation index (SII). The Kaplan-Meier method was used for survival analysis. We conducted univariate and multivariate analyses by using the Cox proportional risk regression model. Software R (version 4.2.0) was used to create the nomogram of prognostic factors. Results The results of the ROC curve analysis showed that the optimal cutoff values of PLR, NLR, and SII were 146.06, 2.67, and 493.97, respectively. The overall response rates were 77.6% and 64.5% in the low and high NLR groups, respectively (P<0.05). The results of the Kaplan-Meier survival analysis revealed that the prognosis of patients in the low PLR, NLR, and SII group was better than that of patients in the high PLR, NLR, and SII group (all P<0.05). The results of the multivariate Cox regression analysis showed that gender, treatment modalities, T stage, and NLR were independent factors affecting the overall survival (OS). In addition, T stage and NLR were independent factors affecting the progression-free survival (PFS) (all P<0.05). The nomogram models of OS and PFS prediction were established based on multivariate analysis. The C-index values were 0.703 and 0.668. The calibration curves showed excellent consistency between the predicted and observed OS and PFS. Conclusion The pretreatment values of PLR, NLR, and SII are correlated with the prognosis of patients with ESCC who underwent radical radiotherapy. Moreover, NLR is an independent factor affecting the OS and PFS of ESCC patients. The NLR-based nomogram model has a good predictive ability.

Objective: To evaluate the effect of 3DBody software assisted problem-based learning (PBL) teaching in orthopedic teaching. Methods: Undergraduates of clinical medicine from grade 2013 who had internship in our hospital were divided into experimental group and control group. Undergraduates in the experimental group were taught by 3DBody software assisted PBL teaching, with designed questions and the method of using 3DBody software handing to students before class. While undergraduates in the control group were taught by traditional teaching, with the use of textbooks and multimedia courseware. Examination scores of theory and probation were compared between the two groups. Questionnaires were used to evaluate the subjective perception of different teaching method among the participants in each group. Results: Scores of theoretical examination in the experimental group (84.6±5.9) were higher than those in the control group (73.2±6.1); scores of probation examination in the experimental group (17.7±2.1) were significantly higher than those in the control group (12.7±1.9); the degree of satisfaction in the experimental group (9.2±0.8) was significantly higher than that in the control group (7.2±1.3); all differences were statistically significant (P<0.05). Conclusion 3DBody software assisted PBL teaching can significantly improve the effectiveness of orthopedic teaching and enhance students' learning initiative and interest, which is worth promoting.

OBJECTIVE: explore the expression of miR-155-5p in Wilms tumor and its effect in regulating the proliferation, migration and apoptosis of Wilms tumor cells. METHODS: Specimens of tumor tissues and paired adjacent tissues were obtained from 40 patients with Wilms tumor for detection of the expression levels of miR-155-5p using RT-qPCR. Wilms tumor cell line G401 was transfected with miR-155-5p mimics and miR-155-5p inhibitor to induce miR-155-5p over-expression and its inhibition, respectively, and the changes in the cell proliferation, migration and apoptosis were assessed using cell counting kit-8 (CCK-8), wound healing assay and fl ow cytometry. RESULTS: RT-qPCR showed that the expression of miR-155-5p decreased significantly in Wilms tumor tissues as compared with normal kidney tissues and was significantly associated with TNM stage ( < 0.05). In G401 cells, over-expression of miR-155-5p significantly inhibited the cell proliferation and migration and promoted cell apoptosis ( < 0.05), and down-regulation of miR-155-5p obviously enhanced the proliferation and migration and suppressed apoptosis of the cells ( < 0.05). CONCLUSIONS miR-155-5p is down-regulated in Wilms tumor and its expression level is correlated with TNM stage. miR-155-5p participates in the progression of Wilms tumor by inhibiting the proliferation and migration and promoting apoptosis of the tumor cells, and may serve as a novel biomarker for diagnosis, therapy and prognostic evaluation of Wilms tumor.
Apoptosis ; Cell Movement ; Cell Proliferation ; Humans ; Kidney Neoplasms ; genetics ; MicroRNAs ; genetics ; Neoplasm Invasiveness ; Wilms Tumor ; genetics