Objective To investigate the effect of ankyrin-repeat domain-containing protein 22(ANKRD22)on the proliferation,invasion,and migration of human hepatoma cells and its molecular mechanism.Methods The TCGA database was used to analyze the expression level of ANKRD22 in normal liver tissue and hepatocellular carcinoma tissue and its association with prognosis.Western Blot and qRT-PCR were used to measure the expression of ANKRD22 in human normal liver cells(L-02)and human hepatoma cells(Huh7,HepG2,MHCC-97H,SK-HEP-1,and SMMC-7721);CCK-8 assay,EdU,wound healing assay,and Transwell assay were used to observe the effect of ANKRD22 on the proliferation,invasion,and migration of hepatoma cells;Western Blot was used to investigate the association of ANKRD22 with cyclins and EMT-related proteins;KEGG and ssGSEA analyses were performed to investigate the mechanism of action of ANKRD22 in hepatoma cells,and related experiments were conducted for validation.The independent-samples t-test was used for comparison of continuous data between two groups;a one-way analysis of variance was used for comparison between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results In the TCGA database,the expression level of ANKRD22 in hepatoma tissue was significantly higher than that in normal liver tissue(t=5.083,P<0.05),and the patients with a high expression level of ANKRD22 had longer overall survival and disease-related survival than those with a low expression level of ANKRD22(P<0.05).The expression level of ANKRD22 in various human hepatoma cell lines was higher than that in human normal liver cells(all P<0.05).Cell proliferation assay showed that the ANKRD22 overexpression group had significantly higher EdU positive rate and proliferation rate than the Vector group(t=19.60 and 6.72,both P<0.001),and compared with the si-NC group,the si-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower EdU positive rate and proliferation rate(all P<0.001).Compared with the Vector group,the overexpression group had significantly higher expression levels of Cyclin E1,Cyclin D1,CDK7,and CDK4(t=3.54,4.95,6.34,and 5.19,all P<0.01),and the si-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower expression levels than the si-NC group(all P<0.001).The overexpression group had a significantly lower expression level of P27 than the Vector group(t=6.12,P<0.001),and the si-ANKRD22#2 group and the si-ANKRD22#3 group had a significantly higher expression level than the si-NC group(both P<0.001).Invasion and migration experiments showed that compared with the Vector group,the ANKRD22 overexpression group had significantly higher migration rate and number of crossings through the membrane(migration group and invasion group)(t=5.01,25.60,and 3.67,all P<0.05),and compared with the si-NC group,thesi-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower migration rate and number of crossings through the membrane(migration group and invasion group)(all P<0.01).The overexpression group had significantly higher expression levels of N-cadherin,Vimentin,and Snail than the Vector group(t=12.13,8.85,and 13.97,all P<0.001),and the si-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower expression levels than the si-NC group(all P<0.001);the overexpression group had a significantly lower expression level of E-cadherin than the Vector group(t=4.98,P<0.01),and the si-ANKRD22#2 group and the si-ANKRD22#3 group had a significantly higher expression level than the si-NC group(both P<0.001).The KEGG enrichment analysis and the ssGSEA analysis showed that ANKRD22 was associated with the PI3K/AKT/mTOR signaling pathway in hepatocellular carcinoma,and the overexpression group had significantly higher expression levels of p-AKT/AKT,p-PI3K/PI3K,and p-mTOR/mTOR than the Vector group(t=12.21,3.43,and 9.75,all P<0.01),and the si-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower expression levels than the si-NC group(all P<0.001).Conclusion ANKRD22 is highly expressed in hepatoma cells and can promote the proliferation,invasion,and migration of hepatoma cells and the activation of the PI3K/AKT/mTOR signaling pathway.

METHODS: Rat BMSCs were isolated, cultured and identified. The time-dependent expressions of TSP-2 and Sox9 in BMSCs under a dynamic mechanical pressure of 0–120 kPa at 0.1 Hz for 1 h were tested by qPCR and Western blotting. The role of TSP-2 in chondrogenic differentiation of BMSCs under mechanical pressure was validated by using small interfering RNA. The impact of TSP-2 and mechanical pressure on chondrogenesis were detected and the downstream signaling molecules were explored using Western blotting. RESULTS: Mechanical pressure stimulation of 0–120 kPa for 1 h significantly upregulated the expression of TSP-2 in BMSCs. The expression of the chondrogenesis markers Sox9, Aggrecan, and Col-II were all upregulated under dynamic mechanical pressure or TSP-2 stimulation. Additional exogenous TSP-2 may potentiate the chondrogenic effect of mechanical stimulation. After knock down TSP-2, the upregulation of Sox9, Aggrecan and Col-II under mechanical pressure was inhibited. The NF-jB signaling pathway responded to both dynamic pressure and TSP-2 stimulation, and the cartilage-promoting effect was blocked by an NF-jB signaling inhibitor. CONCLUSION TSP-2 plays an essential role in the chondrogenic differentiation of BMSCs under mechanical pressure. NF-jB signaling is involved in the mechano-chemical coupling of TSP-2 and mechanical pressure for the chondrogenic differentiation of BMSCs.

Systemic juvenile idiopathic arthritis is one of the common rheumatic and immune diseases in children. It has a sudden onset, obvious systemic symptoms, and lung involvement. However, systemic juvenile idiopathic arthritis with an early manifestation of pulmonary ground-glass opacities combined with macrophage activation syndrome is rare. The clinical data of a child with systemic juvenile idiopathic arthritis with pulmonary ground-glass shadow and macrophage activation syndrome who was admitted to Hubei Maternal and Child Health Care Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology in December 2021 were analyzed retrospectively in order to improve the understanding of rheumatic diseases and pulmonary lesions. The child was admitted to the hospital for 10 days due to rash and fever. Thoracic CT showed scattered ground glass like shadows in both lungs due to the prevention and control screening of COVID-19 pneumonia epidemic situation. After admission, the child was still repeatedly flaccid with high fever, accompanied by dysfunction of both lower limbs. The knee joint MRI found that there was synovitis in the knee joint, and various laboratory indicators suggested macrophage activation syndrome. After that, systemic juvenile idiopathic arthritis was diagnosed. After being treated with methylprednisolone, cyclosporine and topzumab, the clinical remission and the ground-glass shadow of the lung basically disappeared. Through the analysis of this case, it is suggested that clinicians should not ignore other diseases that cause ground glass shadow in the lung during the current epidemic of COVID-19.

Objective:To investigate the correlation between sleep quality and risk of female complicated vulvovaginal candidiasis (VVC).Methods:From January 2021 to June 2021, patients in the gynecological clinic of Xiangya Hospital of Central South University were continuously enrolled as the research objects using a cross-sectional survey. A self-made questionnaire was used to collect the age, marital status, education level, family monthly income, place of residence in the past two years, maternity history, number of births, intrauterine device, number of abortions, frequency of sex life, use of contraceptives within two months, use of antibacterial drugs within two weeks. Generalized Anxiety Disorder Scale, Patient Health Questionnaire, Health Questionnaire Somatic Symptom Group Scale, Pittsburgh Sleep Quality Index were used to collect patients′ anxiety, depression, somatization symptoms, and sleep quality conditions. The total scores of sleep quality and the scores of each dimension were used as observation indicators. Three logistic regression analysis models were constructed to explore the relationship between sleep quality and complicated VVC groups.Results:Patients in the complex VVC group were significantly higher in age, married, middle school education, rural area of residence in the last two years, birth history, number of births ≥3, sexual frequency≥1/week, and no antibiotic use within two weeks compared to those in the control group (all P<0.05). Without adjusting for confounding factors, women with poor subjective sleep quality had a 6.73-fold increased risk ( OR=7.73, 95% CI: 3.22-18.55) of complex VVC compared with those with good subjective sleep quality. After adjusting for confounding factors, the risk was further increased to 9.08 fold ( OR=10.08, 95% CI: 3.47-29.33)(all P<0.05). Compared with women without sleep disorders, women with mild sleep disorders had a 97% increased risk of complex VVC ( OR=1.97, 95% CI: 1.15-3.37). After adjusting for confounders, the risk remained 97% higher ( OR=1.97, 95% CI: 1.10-3.55)(all P<0.05). Conclusion:Poor subjective sleep quality and mild sleep disorder may be associated with the risk of complex VVC.

Objective:To explore the value of metal artifact reduction in oblique sagittal multi-acquisition with variable resonance image combination slab selectivity (MAVRIC-SL) inversion recovery (IR) sequence compared with conventional fast spin-echo (FSE) fat saturation (FS)-T 2WI and proton density weighted imaging (PDWI) on the postoperative review of patients with metal-fixed anterior cruciate ligament reconstruction (ACLR) at 3.0 T MR. Methods:From June to November 2018, 27 patients with metal fixtures in ACLR were prospectively recruited in the First Hospital of China Medical University. The FSE sequences (FS-T 2WI and PDWI) and the MAVRIC-SL IR were performed at 3.0 T MRI system. The maximum artifact slice was selected and ROI was drawn, then the implant and artifact area and the number of artifact-influence slices were recorded. Image signal-to-noise ratio (SNR) was calculated. The degree of image artifacts (artifact degree of spiked ligament staple and its influence on the diagnosis of surrounding structures), measurability (measurability of distance between the upper edge of the staple and the lower edge of the graft entrance on tibia), and other degree of artifacts (whether there were metal artifacts other than implants that affect the observation of other structures within the scanning range) were scored. The Friedman M test and pairwise comparison were used to compare and analyze the quantitative and qualitative data. Results:The differences in artifact area, the number of artifact-influence slices and SNR among FS-T 2WI, PDWI and MAVRIC-SL IR images were statistically significant (all P<0.001). Pairwise comparison analysis suggested that the metal implant artifact area and the number of artifact-influence slices in the MAVRIC-SL IR image were smaller, and the SNR was higher than those of FS-T 2WI and PDWI (all P<0.05 after correction). The differences of the three scores among FS-T 2WI, PDWI and MAVRIC-SL IR were statistically significant (all P<0.001). Pairwise comparison analysis suggested that the degree of image artifacts score of MAVRIC-SL IR sequence was lower, and the measurability and other degree of artifacts scores were higher than FS-T 2WI and PDWI (all P<0.05 after correction). Conclusion:MAVRIC-SL can significantly reduce the metal artifacts and improve image quality in the postoperative MRI review of the patients with metal-fixed ACLR. Therefore, oblique sagittal MAVRIC-SL IR sequence can be used for assisting the diagnosis of conventional FS-T 2WI and PDWI.

Objective To learn the needs of family physicians for tertiary hospital support in the hierarchical medical system. Methods In July 2017 cluster sampling was made to 135 family physicians of Changning district of Shanghai for a " Family physicians needs questionnaire" . It covered such aspects as their demographic characteristics and their needs. The data so acquired were subject to descriptive and logical analysis. Results 99.3% of them need preferential appointment of specialists at tertiary hospitals;93.3% of them need preferential appointment of examinations; 82.2% of them need preferential outpatient visits; while 83.7% of them need preferential hospitalization and surgery scheduling. In terms of the approaches for preferential appointment of specialists, 84.4% of them prefer specialist appointment at short notice, and 73.3% of them need direct online appointment for large-scale device examinations at tertiary hospitals. In terms of mentoring scenarios, 62.2% of them prefer mentoring during outpatient rounds at fixed schedules of specialists in their community. Conclusions Tertiary hospitals are recommended to establish green pathways for referrals and priority measures in order to improve capabilities of family physicians and the shared platform in the hierarchical medical network. They are also expected to guide family physicians in their research paper writing, thus fully supporting the hierarchical medical system.

Objective To explore the effect and compare the relapse rates of 1-desamino-8-D-Arginine Vasopressin(DDAVP) different withdrawal ways after initial 3 months in primary monosymptomatic nocturnal enuresis(PMNE) patients,in order to provide some evidences and references to use DDAVP to cure PMNE preferably.Methods Two hundred and fifty-six cases PMNE patients who were treated in Hubei Maternal and Child Health Hospital from November 2014 to June 2016 were selected and randomly divided into group A (DDAVP immediate withdrawal group,65 cases),group B(DDAVP day reduction group,58 cases) and group C (DDAVP step reduction group,60 cases).All patients were given DDAVP tables for 3 months.After 3 months for DDAVP,patients who were effective (full respond and partial respond) to DDAVP continued to undergo a withdrawal stage,those in group A underwent immediate cessation,those in group B continued to receive the effective dose every other day for 2 months and those in group C were step by step tapered by 0.05-0.10 mg every 2-4 weeks until completely stopped,the period was not more than 3 months.All patients had a follow-up visit for 3 months after cessation of DDAVP.Results A total of 183 patients completed the study finally,there were 65 patients in group A,58 patients in group B and 60 patients in group C.Initial 3 months the effective rates of group A,B and C were respectively 89.23％ (58/65),89.66％ (52/58) and 86.67％ (52/60),there were not statistically significant difference(x2 =0.309,P =0.857).There were 58 patients in group A,52 patients in group B and 52 patients in group C continued to undergo the withdrawal stage.One month after cessation of DDAVP,the effective rates of group B (88.46％,46/52) and group C (92.31％,48/52) were significantly higher than group A(67.24％,39/58) (x2 =7.030,P=0.008;x2 =10.417,P=0.001),while the relapse rates of group B(19.23％,10/52) and group C(17.31％,9/52) were significantly less than group A(36.21％,21/58) (x2=3.904,P=0.048;x2=4.937,P =0.026).Three months after cessation of DDAVP,the effective rates of group C (78.85％,41/52) were significantly higher than group A (50.00％,29/58) and group B (57.69％,30/52) (x2 =9.859,P=0.002;x2 =5.371,P=0.020),and the relapse rates of group C(32.69％,17/52) were significantly less than group A (55.17％,32/58) and group B (51.92％,27/52) (x2 =5.609,P =0.018;x2 =3.939,P =0.047),while the effective rates and relapse rates were not statistically significant difference between group A and group B(x2 =0.652,P =0.419;x2 =0.116,P =0.733).Severe adverse events related to DDAVP were not observed in any patients.Conclusion Gradual withdrawal after initial 3 months of DDAVP may improve the effect and reduce the relapse rates,the short-term and long-term curative effects of step-by-step withdrawal treatment are both well,while long-term curative effects of every other day withdrawal treatment is not obviously well.

Objective To assess the clinical walue of 3D retroperitoneal laparoscopic renal pedicle disconnection by comparing its clinical effects with those of traditional 2D laparoscopic.Methods Clinical data about 49 cases from our department with chyluria respectively treated by 3D retroperitoneal laparoscopic and 2D laparoscopic renal pedicle disconnection were analyzed retrospectively.They were divided into 3D group(26 cases)and 2D group(23 cases),and operated by the same salty doctor.The operation time,intraoperative blood loss,postoperation hospital stay and recurrence rate of the two group were compared.Results The operation time in 3D group[(95.2±30.3)min] was shorter than 2D group[(120.4±25.8)min,P<0.05].The intraoperative blood loss in 3D group[(30.0±12.7)ml] was less than 2D group[(55.0±21.7)ml,P<0.05].There were no difference in postoperation hospital stay between the two groups.All of the cases were not recurred.Conclusion 3D Retroperitoneoscopic renal pedicle lymphatic disconnection for treatment of chyluria is safty and feasible.3D laparoscopy provides high-quality 3D endoscopic view and facilitates precise manipulation during surgery,resulting in shorter operation time and intraoperative blood loss compared with 2D laparoscopy.

Objective To investigate the expression and clinical significance of H1 receptor in kidney and bladder tissue of rats after long term ketamine intraperitoneal injection.Methods This study was conducted from May 2012 to December 2012.Sixty male 2-month-old SD rats,weighted (200±10) g,were randomly divided into Group A and Group B.Each group concluded 30 rats.In the Group A,Ketamine (100 mg/kg) was given as intraperitoneal injection every other day,while normal saline (100 mg/kg) was given in Group B.The dosage was adjusted every week according to the weight of rats.After 2,4 and 6 months,10 rats from each group were randomly chosen.First,the micturition number during 2 h was recorded.Then,urine samples over a 24 h period were collected and the content of Na+ and K+ were determined.Finally,the blood samples were obtained from the apex of heart for the creatinine determination.The kidneys and bladders were harvested after the rats were sacrificed.HE staining was conducted on all the tissues.Immunohistochemical S-P method was used to detect the expression of H1 receptor in the bladder and kidney tissues from Group A and Group B.The average optical density (A Value) in each group was separately calculated by Imagepro-plus 6.0 software.All the parameters,mentioned above,were carefully compared.Results The successive rate of establishing rats model was 90％ (9/10),according to the pathological result after 6 months injection.The urine volume of 24 h in group A and B were (15.9±1.3) and (10.1±0.8) ml,respectively.Micturition frequency during 2 hours in group A and B were (6.9±1.4) and (3.0±0.5) times.The urine volume of 24 h and micturition frequency during 2 hours were significantly increased in group A (P＜ 0.05).The urine sodium within 24 h in group A was (1.7±0.1) mmol,which is increased significantly than that in group B (1.0±0.1 mmol).While the urine potassium was less in group A (1.1±0.1 mmol/d) than in group B (2.6±0.1 mmol/d) (P＜0.05).But the serum creatinine level were (60.5±6.8) and (58.1± 3.9) μmol/L in group A and B,which had no difference between the two groups (P＜0.05).The expression of H1 receptor in kidneys and bladder in group A was significantly raised compared with group B (P＜0.05).In the group A,the expression of H1 receptor level in kidney was 0.008±0.001,0.016±0.001,0.023±0.004 after 2,4 and 6 months drug used.The expression level in group A were significantly difference than that in group B (0.003±0.001,0.004±0.002,0.003±0.001) (P＜0.05) and goes up with prolonging the drug using.While in the bladder tissue,the level of H1 receptor expression was 0.017±0.006,0.031±0.012,0.036±0.007 in group A and 0.015±0.007,0.016±0.005,0.016±0.004 in group B,which could be noticed a significantly increasing in group A (P＜0.05).In 4 and 6 months,the H1 receptor expression level significantly raised than that in 2 months (P＜0.05).Conclusions Long term ketamine addiction exerts toxicity not only on the bladder but also on the kidney.The increased expression of H1 receptor in rats' kidney and bladder tissues of group A indicates that H1 receptor may be related to the ketamine-associated urinary system dysfunction.The urine sodium and potassium within 24 h may be a sensitive index for the assessment of degree of kidney damage in the early stage of ketamine-induced dysfunction than serum creatinine.

Objective To investigate the expression and clinical significance of four histamine receptors:H1R, H2R, H3R and H4R in urinary bladder of patients with ketamine-induced cystitis. Methods Immunohistochemical S-P method was used to detect the expression levels of histamine receptors:H1R, H2R, H3R and H4R in bladder tissues of 10 patients with ketamine-induced cystitis (experimental group) and distal tissue away from bladder tumors of 10 patients with cystectomy (control group). The average optical density (OD) values of four kinds of different histamine receptors were separately calcu-lated by Imagepro-plus 6.0 in two groups. At the same time, mast cells were marked by toluidine blue special dyeing and were counted. Results Comparing with control group, the expression levels of H1R, H2R, and H4R were significantly in-creased in experimental group (P＜0.05). Mast cells diffused interstitial bladder infiltrates (P＜0.05). There was no signifi-cant difference in the expression of H3R in two groups. Conclusion Mast cells, H1R, H2R, and H4R are closely related to the ketamine-induced cystitis, which may be new diagnostic indicators and new treatment targets of ketamine-induced cysti-tis.