Objective To evaluate the impact of small interference RNA (siRNA)-induced silencing of adrenomedullin (ADM) gene on the growth of and apoptosis in a malignant melanoma cell line A375.Methods Three siRNAs targeting ADM gene were constructed and transfected into A375 cells.Real-time quantitative reverse transcription PCR (qRT-PCR) was performed to measure the expression of ADM to select the most efficient siRNA for the next experiment.A375 cells were classified into 3 groups to be transfected with the selected siRNA (test group),non-specific sequences (negative control group),or remain untransfected (blank control group).Then,an immunohistochemical SP method was used to determine the expression of ADM at 24hours,methyl thiazolyl tetrazolium (MTT) assay to detect the proliferation of A375 cells at 24 and 48 hours,flow cytometry to observe cell apoptosis at 48 hours.Results qRT-PCR showed that ADM-siRNA-2 was the most efficient.The expression of ADM was significantly lower in the test group than in the other two groups.The proliferation level (represented as the absorbance value at 490 nm) of A375 cells was 0.389 ± 0.0375 and 0.469 ± 0.0330 in the test group at 24 and 48 hours respectively,significantly lower than that in the negative control group (0.548 ± 0.0376,0.676 ± 0.0374,both P＜ 0.05) and blank control group (0.574 ± 0.0733,0.685 ± 0.0154,both P ＜ 0.05).Flow cytometry revealed a statistical increase in early apoptosis rate in the test group compared with the blank control group and negative control group ((20.200 ± 6.046)％ vs.(1.667 ± 0.340)％ and (4.587 ± 1.294)％,both P ＜ 0.05).No significant differences were observed in the proliferation level or early apoptosis rate between the negative control group and blank control group (both P ＜ 0.05).Conclusion The siRNA-induced silencing of ADM gene can inhibit the proliferation of,but promote the apoptosis in,A375 cells.

Background Orbital fracture in children has its own characteristics.Choosing the suitable plastic materials is the effective measurement to improve the successful rate of surgery.Absorbable plate filling provides a new material for children with orbital fracture.However,few studies compare the effectivity and safety between absorbable plate filling and conventional filling.Objective The aim of this study was to compare the longterm efficacy and safety between absorbable plate and Medpor implants in the repair of pediatric orbital fracture.Methods Clinical data of 72 eyes of 72 pediatric patients with orbital fracture who received surgicals reconstruction in Beijing Tongren Eye Center from 2008 April to 2014 August were respectively analyzed,including 28 patients with absorbable plates filling and 44 patients with Medpor implants.Demography and preoperative clinical signs were matched between the two groups.Operative procedure was same except the implanted materials.The functional training of extraocular muscle was performed since the second day after surgery in both groups.The imaging findings of the orbit were evaluated by computed tomography (CT) before and after surgery.The patients were followed-up for avarage 12.7 months after operation.The therapeutic effectiveness and safety were compared between the different implants,such as incidence of enophthalmos,extraocular movement impairment and diplopia,and the sensory deficit of skin and complications were documented.Results Extraocular muscle movement improved and diplopia alleviated or disappeared in 92.9％ (26/28) in the absorbable plate group and 90.9％ (40/44) in the Medporgroup,without significant difference between the two groups (P=0.57).The sensory deficit of skin was found in 13 eyes (46.4％) and 20 eyes (45.5％ in the absorbable plate group and Medpor group,respectively,and there was no significant difference between the two groups (P =0.56).CT revealed that the anatomical reconstruction of orbits could be achieved in all patients in both groups without the shifting of implants and extrusion.No enophthalmos and postoperative infection were found in absorbable plate group,however,the residual enophthalmos or late infection was found in I eye for each in the Medpor group.Conclusions Both the absorbable plate and Medpor implants show good results for reconstruction of pediatric orbital fracture.Absorbable plate is an ideal material for pediatric orbital blowout fracture because of fewer postoperative complications.

Objective To explore the variation trend of natural killer (NK) cell subsets in the recipients infected with cytomegalovirus (CMV) after renal transplantation. Methods Clinical data of 92 renal transplant recipients were retrospectively analyzed. All recipients were divided into the CMV infection group (n=43), CMV infection recovery group (n=13), stable renal function group (n=15), rejection group (n=11) and other infection group (n=10). In addition, healthy adult volunteers were enrolled in the healthy control group (n=15). The proportion of NK cells in peripheral blood, the expression proportion and the mean fluorescence intensity (MFI) of CD226 and CD16 in NK cells were observed and statistically compared among different groups. Results The proportion of NK cells was 4.9% (2.2%, 11.5%) in the CMV infection group and 3.7% (2.3%, 6.5%) in the CMV infection recovery group, which were significantly lower than those in the other groups (all P < 0.05). The expression proportion of CD226 and CD16 in NK cells in the CMV infection group was significantly lower compared with those in the healthy control group and stable renal function group(all P < 0.05). The expression proportion of CD226 and CD16 in NK cells in the CMV infection recovery group was remarkably higher than those in the CMV infection group (both P < 0.05). The MFI of CD226 and CD16 in the CMV infection group was significantly lower than those in the healthy control group (both P < 0.05). The MFI of CD226 and CD16 in the CMV infection recovery group was significantly higher than those in the CMV infection group (both P < 0.05). Conclusions The expression proportion and MFI of CD226 and CD16 in NK cells are down-regulated in CMV infection period, whereas up-regulated during the CMV infection recovery period, prompting that CD226 and CD16 expressed by NK cells are intimately correlated with the course of CMV infection.

Objective:To observe the clinical effects of intraperitoneal perfusion of bevacizumab combined with albumin paclitaxel and carboplatin in the treatment of malignant peritoneal adhesion caused by ovarian cancer.Methods:From January 2016 to December 2020, 54 patients treated in our hospital with malignant peritoneal adhesions caused by ovarian cancer were enrolled in this study. They were randomly divided into experimental group ( n=27) and control group ( n=27) according to the random number table method. The treatment regimen of the experimental group was intravenous infusion of albumin paclitaxel plus intraperitoneal infusion of carboplatin and bevacizumab. The treatment regimen of the control group was intra-venous infusion of albumin paclitaxel plus intraperitoneal infusion of carboplatin. The treatment was repeated every 21 days, and the therapeutic effect was evaluated every two cycles. The treatment lasted for six cycles. The efficacy and incidence of adverse reactions were compared between the two groups. Results:The remission rate of incomplete malignant bowel obstruction of the experimental group was higher than that of the control group [85.19% (23/27) vs. 59.26% (16/27)], the total effective rate of the experimental group was higher than that of the control group [74.07% (20/27) vs. 44.44% (12/27)], and there were statistically significant differences ( χ2=4.523, P=0.033; χ2=4.909, P=0.027). After treatment, the levels of vascular endothelial growth factor (VEGF) in ascites of the experimental group and the control group were significantly lower than those before treatment [(80.33±1.41) pg/ml vs. (310.45±3.35) pg/ml, t=449.884, P<0.001; (135.68±1.60) pg/ml vs. (310.46±3.09) pg/ml, t=499.281, P<0.001], and after treatment, the VEGF level in the experimental group decreased more significantly than that in the control group ( t=-134.907, P<0.001). Patients in the experimental group and the control group tolerated the treatment well, and there were no significant differences in the incidences of adverse reactions such as hypertension (11.11% vs. 3.70%, χ2=0.270, P=0.603), neutropenia (14.81% vs. 11.11%, χ2<0.001, P>0.999), peripheral neuropathy (3.70% vs. 0, χ2<0.001, P>0.999), diarrhea (7.41% vs. 3.70%, χ2<0.001, P>0.999), nausea (3.70% vs. 0, χ2<0.001, P>0.999), epistaxis (7.41% vs. 0, χ2=0.519, P=0.471) or albuminuria (3.70% vs. 0, χ2<0.001, P>0.999) between the two groups. Conclusion:Intraperitoneal perfusion of bevacizumab combined with chemotherapy is superior to simple chemotherapy in the treatment of malignant peritoneal adhesion caused by ovarian cancer.

Objective:To investigate the expression of forkhead box protein O3(FOXO3) in pancreatic cancer and its effect on the motility and proliferation of pancreatic cancer cells.Methods:The FOXO3 expression in pancreatic cancer and adjacent tissues was retrieved from LinkedOmics database. Western blotting and real-time quantitative polymerase chain reaction were used to detect FOXO3 expression in pancreatic cancer cells and human pancreatic stellate cells. PANC-1 and MIAPaCa-2 pancreatic cancer cells with low FOXO3 expression were selected to transfect FOXO3 overexpression plasmid and negative control plasmid, respectively. The motility and proliferation ability of pancreatic cancer cells were detected by colony formation assay, cell scratch assay, Transwell assay and flow cytometry.Results:In the LinkedOmics database, the relative expression of FOXO3 protein in the cancer tissues of 64 patients with pancreatic cancer was significantly lower than that in the adjacent tissues ( t=8.36, P<0.001). The number of clones in PANC-1 cell line was (30.0±6.6) after overexpressed FOXO3, which was lower than that in negative control cells (92.7±6.7), and the difference was statistically significant ( t=11.54, P<0.001). After overexpressed FOXO3 in PANC-1 and MIAPaCa-2 cell lines, the scratch repair rate was significantly decreased compared with the control group. In Transwell experiment, the number of cells in FOXO3 overexpressed group in PANC-1 cell lines was (21.0±6.6), which was lower than that of negative control groups (55.7±8.5), and the difference was statistically significant ( t=5.59, P=0.005). The results of MIAPaCa-2 cell line were consistent with that of PANC-1 cell line. After overexpressing of FOXO3 in PANC-1 and MIAPaCa-2 cell lines, the proportion of cells in the G0/G1 phase decreased, while the proportion in the S phase increased. Conclusion:The expression of FOXO3 was decreased in pancreatic cancer. Overexpression of FOXO3 could significantly inhibit the proliferation, migration and invasion of pancreatic cancer cells and induce cell cycle arrest, which is a potential target for the treatment of pancreatic cancer.

Objective:To investigate the expression of IKBKE and NF-κB in pancreatic cancer, and to explore the effect of IKBKE on pancreatic cancer proliferation and migration.Methods:Immunohistochemistry staining was used to study the expression of IKBKE and NF-κB in tissues of 61 pancreatic cancer patients admitted to the Second Hospital of Tianjin Medical University from January 2012 to January 2017 and 13 normal pancreatic tissues. The correlations between those expression to clinicopathological features were analyzed. Lentivirus mediated RNAi was transfected into pancreatic cancer cells to block IKBKE. Western blot was performed to test the silencing effeciency; CCK-8 and plate clone and scratch assays were used to investigate the proliferation and migration of pancreatic cancer.Results:Immunohistochemical staining showed that 60 (98.4%) of IKBKE staining were weakly positive, positive, and strongly positive in pancreatic cancer tissues, which were significantly higher than normal pancreatic tissues(76.9% cases were weakly positive and the rest were negative), and the differences were statistically significant ( P<0.05). All cases of NF-κB exhibited weakly positive expression and above in pancreatic cancer tissues, which was markedly higher than normal tissues (30.8% cases were weak positive and the rest were negative staining), statistically significant ( P<0.05). Survival analysis showed that patients with high level of IKBKE showed a shorter overall survival ( P<0.05). CCK-8, plate cloning and scratch assays showed that the proliferation and migration of were significantly decreased in IKBKE knocking down group ( P<0.05). Conclusions:IKBKE and NF-κB are highly expressed in pancreatic cancer, and IKBKE is correlated with NF-κB in pancreatic cancer. Blocking of IKBKE could distinctly inhibit the proliferation and migration of pancreatic cancer.

Objective To summarize our experience in the diagnosis and treatment of hepatic epithelioid angiomyolipoma (HEAML),with the aim to reduce the future misdiagnosis rate.Methods The PubMed,Medline,China Science Periodical Database (CSPD),and VIP Databases were searched from January 2000 to March 2018 on all reports on HEAML.Results There were 409 cases of HEAML in 97 reports.The ratio of men to women was 1∶4.84.The age ranged from 12 to 80 years and the median age was 44 years.61.9％ of patients (205/331) were asymptomatic,while 34.7％ (115/331) had upper or right upper quadrant abdominal discomfort.Some patients presented with abdominal mass,gastrointestinal reaction,low grade fever or weight loss.The clinical symptoms in 78 patients were not mentioned in the reports.The misdiagnostic rate of HEAML was as high as 40.3％ (165/409).The imaging findings of HEAML were nonspecific.Ultrasound,CT and MRI scan usually showed contrast enhancement in the arterial phase.Most lesions were accompanied by central vessels with early drainage veins.The enhanced scans showed varied characteristics.The ratios of fast wash-in and fast wash-out,to fast wash-in and slow wash-out,and to delayed enhancement were roughly 4∶ 5∶ 1.A definitive diagnosis of HEAML is based on the pathological findings of epithelioid cells in the lesions and the expressions of HMB45,SMA,Melan-A and Actin on immunohistochemical staining.HEAML had a relatively low malignant rate of 3.9％.Surgical resection was the main treatment for HEAML.Conclusion HEAML was a rare and easily misdiagnosed disease.,which could be diagnosed by taking into account the clinical course,imaging,pathological and immunohistochemical findings.HEAML.

Objective:To examine the association between height loss and calcaneus bone mineral density (BMD) through data gathered from the China Kadoorie Biobank (CKB).Methods:The present study included 24 231 participants who attended the CKB resurvey during 2013-2014, in which calcaneus BMD was measured by quantitative ultrasound method for the first time. Height loss was calculated according to the differences appeared in height measurement between baseline and resurvey. We used linear regression models to estimate the association between height loss and BMD measures.Results:The mean interval between baseline and resurvey was (8.0±0.8 ) years. 33.0 % of the participants showed a height loss of ≥1.0 cm, and another 3.7 % were with height loss of ≥3.0 cm. After adjustment for potential confounders, there was a linear correlation seen between height loss and BMD ( P for all linear trend were <0.001). The βs (95 %CIs) for each 1.0 cm of height loss were -0.79 (-0.95--0.63) for broadband ultrasound attenuation (BUA), -2.74 (-3.35--2.13) for speed of sound (SOS), and -1.29 (-1.54--1.04) for stiffness index (SI). Compared with participants with stable height, the multivariate-adjusted βs (95 %CIs) for those with height loss of ≥3.0 cm were -3.29 (-4.08--2.50) for BUA, -10.70 (-13.66--7.73) for SOS, and -5.16 (-6.36--3.96) for SI, respectively. According to the subgroup analyses, the association of height loss with BMD measures seemed to be more apparent among females, in those aged ≥55 years, and those being less physically active. Conclusions:BMD became lower with the increase of height loss. Regular height measurement may contribute to the early diagnosis and prevention of osteoporosis.

Objective:To evaluate the transitions of frailty status and related factors influencing its worsening in middle-aged and elderly adults.Methods:Data was obtained from the Beijing MJ Health Screening Center. A total of 13 689 participants who attended health checkups at least twice during 2008-2019 and had more than three years' intervals during these two health checkups were included in the study. The frailty index comprising 28 variables was used to measure frailty status. Frailty was defined as frailty index ≥0.25, and prefrailty was defined as frailty index >0.10 and <0.25. Logistic regression analysis was performed to investigate the association of socio-demographic factors and lifestyle characteristics with the worsening of frailty status, stratified by frailty status at the first health checkup.Results:The mean age at the first and last health checkups were (42.3±9.2) and (47.9±9.3) years, respectively. The mean interval during these two health checkups was (5.7±1.9) years. At the first health checkup, the prevalence of frailty and prefrailty were 2.5% and 50.3%, respectively. While at the last health checkup, the prevalence of frailty and prefrailty rose to 3.9% and 55.4%. Of all participants, 67.3% remained in the same frailty state, 21.2% worsening, and 12.5% improving. In robust participants at the first health checkup, older age, female, low education level, smoking cessation, daily smoking, being general obesity measured by BMI or central obesity measured by WHR showed an increased the risk of worsening frailty status. However, in prefrail participants at the first health checkup, older age, female, general, or central obesity presented as risk factors for worsening frailty status.Conclusion:Modifiable factors such as low education level, smoking, and obesity may increase the risk of worsening frailty status.

Objective:To compare the consistency of frailty status measured by Fried phenotype and frailty index composed of different numbers of deficits, and their prospective associations with risk of mortality.Methods:Data of 23 615 participants from the second resurvey of the China Kadoore Biobank (CKB) was used. Fried phenotype was constructed using five phenotypes, and frailty indexes (FI) were constructed using 28 and 40 deficits, respectively. We calculated the Weighted Kappa coefficient to compare the consistency of three measures in the classification of frailty status. Cox regression was performed to analyze the association of frailty status with risk of mortality.Results:The frailty prevalence calculated by Fried phenotype, FI-28, and FI-40 were 5.4%, 7.9%, and 4.0%, respectively. The Kappa coefficients of Fried phenotype with FI-28 and FI-40 were 0.357 and 0.408, respectively. The Kappa coefficients of FI-28 and FI-40 was 0.712. During an average of (3.9±0.5) years of follow-up, 755 participants died. When Fried phenotype was used, compared with the robust participants, the prefrail and frail participants had increased risk of mortality, the multivariable-adjusted HRs were 1.60 (95% CI: 1.32-1.94) and 2.90 (95% CI: 2.25-3.73), respectively. When FI-28 was used, the corresponding HRs were 1.71 (95% CI: 1.39-2.11) and 2.52 (95% CI:1.95-3.27) for prefrail and frail participants, and when FI-40 was used, the corresponding HRs were 1.98 (95% CI:1.60-2.44) and 3.71 (95% CI: 2.80-4.91). The association of frailty status with mortality differed in different age groups, with the association stronger in younger adults than in older adults. Conclusion:Fried phenotype and frailty index constituted with different numbers of deficits showed good consistency; which can be used to well predict the risk of mortality.