Objective:To analyze the expression of silent information regulator 2-related enzyme 1 (SIRT1) and its relationship with chondrocyte apoptosis in patients with Kashin-Beck disease (KBD).Methods:Twenty patients with KBD were selected as the KBD group from Guide County, Qinghai Province, and 40 healthy subjects matched by age and sex were selected as the control group. Fasting elbow venous blood of the study subjects was collected, and peripheral blood mRNA levels of SIRT1 and selenoprotein genes [glutathione peroxidase (GPX) 2, GPX3, thioredoxin reductase (TXNRD) 1, TXNRD3, iodothyronine deiodinase Ⅰ (DIO1), and selenophosphate synthetase 2 (SPS2)] were detected by real-time PCR. The correlation between SIRT1 expression and selenoprotein genes in peripheral blood of KBD patients was analyzed by curve fitting method. Meanwhile, normal human chondrocytes cultured in vitro were divided into control group (without any treatment), resveratrol (RES) group (to verify the activation effect of RES on SIRT1), tert-butyl hydroperoxide (tBHP) injury group (oxidative injury model of chondrocyte), and RES protection group (tBHP injury after RES pre protection). The mRNA levels of SIRT1, selenoprotein genes, and apoptosis-related genes [B lymphoblastoma-2 gene (BCL2), BCL2-associated X protein (BAX), nuclear transcription factor κB (NF-κB) p65, and tumor suppressor gene P53] in each group of cells were detected by real-time PCR. Results:In the population study, the peripheral blood SIRT1 mRNA level in the KBD group (1.12 ± 0.38) was lower than that of control group (1.87 ± 0.97), and the difference was statistically significant ( t = 3.31, P = 0.002). According to curve fitting analysis, the mRNA levels of GPX3, TXNRD1, and TXNRD3 in peripheral blood of KBD group increased with the increase of SIRT1 mRNA level ( R2 = 0.48, 0.66, 0.95, P < 0.001). The level of DIO1 mRNA showed a trend of decreased first and then increased with the increase of SIRT1 mRNA level ( R2 = 0.51, P = 0.024). The mRNA levels of GPX2 and SPS2 showed no significant change trend with the increase of SIRT1 mRNA level ( R2 = 0.16, 0.12, P = 0.064, 0.114). In cell studies, compared with the control group (1.00 ± 0.10), the SIRT1 mRNA level in the RES group (1.79 ± 0.07) was higher ( P < 0.05). Compared with tBHP injury group, the RES protection group had higher mRNA levels of selenoprotein genes GPX3, TXNRD1, TXNRD3, and DIO1 ( P < 0.05); the mRNA levels of apoptosis-related genes BAX, P53 and the ratio of BAX/BCL2 were lower, while the mRNA levels of BCL2 and NF-κB p65 were higher ( P < 0.05). Conclusions:KBD patients have low expression of SIRT1. And RES activation of SIRT1 may enhance the antioxidant capacity of chondrocyte by up-regulating the expression of selenoprotein genes, thus inhibiting chondrocyte apoptosis.

  Objective  To investigate the occurrence of chronic postsurgical pain (CPSP) among patients with preoperative COVID-19, and further analyze the risk factors for CPSP.  Methods  This study was a ambispective cohort study, with subjects from a completed cohort study with follow-up. We included the clinical data of the patients with preoperative COVID-19 who underwent surgery at Peking Union Medical College Hospital from December 1, 2022 to February 28, 2023. Follow-up was conducted up to 6 months postoperatively, with the primary outcome being CPSP. Multivariate Logistic regression analysis was used to analyze the correlation between COVID-19-related exposure indicators and CPSP.  Results  A total of 4117 surgical patients were included, all of whom had preoperative COVID-19. Among them, 4002 cases had mild symptoms during the acute phase, 62 cases had severe symptoms, and 53 cases were critically ill. At 6th month postoperatively, 1298 cases (31.53%) had long COVID-19 syndrome, and the incidence of CPSP was 5.59% (95% CI: 4.88%-6.28%). After adjusting for confounding factors including age, gender, comorbidities, anesthesia method, and type of surgery, multivariate Logistic regression analysis revealed that critically ill COVID-19 during the acute phase (aOR=3.35, 95% CI: 1.48-7.62, P < 0.001) and presence of long COVID-19 syndrome postoperatively (aOR=2.50, 95% CI: 1.90-3.29, P < 0.001) were associated with CPSP.  Conclusions  It is clear for the first time that critically ill COVID-19 during the acute phase and the presence of long COVID-19 syndrome postoperatively are the risk factors for CPSP among patients with preoperative COVID-19.

A rapid determination of methamphetamine, amphetamine, 6-monoacetylmorphine, and morphine in hair samples by UPLC-MS/MS was established and optimized.The concentration of target compounds in the hair of drug abusers and drug laboratory technicians was investigated and the cut-off value was evaluated.After cleaned hair was extracted by grinding with methanol-water (7∶3) at 3 000 r/min for 100 s, the final solution after adjusting the volume to methanol-water (1∶1) was analyzed by UPLC-MS/MS.The analytes were gradient eluted on a Waters Acquity BEH C18 (2.1 mm × 100 mm, 1.7 μm) column with 5 mmol/L ammonium formate-0.1% formic acid aqueous solution and acetonitrile as mobile phase at a flow rate of 0.4 mL/min. The ESI+ ion source and multiple reaction monitoring (MRM) were used to select the qualitative and quantitative ion pairs of the four target compounds. All analytes showed good linearity (R2 ≥ 0.999 6) in the range of 0.01-5 ng/mg (except amphetamine in 0.01-4 ng/mg), limit of the quantitation was 0.01 ng/mg, and the limit of detection was 0.001-0.008 ng/mg.The accuracy, precision, matrix effect, and recovery all met the requirements of biological sample methodology.According to the comprehensive consideration of the receiver operating characteristic (ROC) curve, Youden index, law enforcement cost and intensity, the reference cut-off values were methamphetamine ≥ 0.1 ng/mg; amphetamine ≥ 0.025 ng/mg; 6-monoacetylmorphine ≥ 0.05 ng/mg; morphine ≥ 0.05 ng/mg.The method established in our research can quickly and accurately detect the contents of methamphetamine, amphetamine, 6-monoacetylmorphine, and morphine in hair.This study provides some reference for the public security system to make more rational cut-off values in the norm of drug-related personnel hair samples detection in the future.

Purpose: This study aimed to compare the ability of B-mode ultrasonography and magnetic resonance imaging (MRI) to predict the repairability of large-to-massive rotator cuff tears (RCTs). Methods: This cross-sectional study included participants with large-to-massive RCTs who underwent arthroscopic repair. B-mode ultrasonography and MRI were conducted prior to arthroscopic repair. B-mode ultrasonography was used to evaluate the echogenicity of the rotator cuff muscle using the Heckmatt scale. Intra-rater and inter-rater reliabilities were examined for two independent physicians. MRI was used to evaluate the degrees of tendon retraction, fatty infiltration of rotator cuff muscles, and muscle atrophy. Finally, two experienced orthopedic surgeons performed surgery and decided whether the torn stump could be completely repaired intraoperatively. Results: Fifty participants were included, and 32 complete repairs and 18 partial repairs were performed. B-mode ultrasonography showed good intra-rater reliability and inter-rater reliability for assessment of the muscle echogenicity of the supraspinatus and infraspinatus muscles. The correlation coefficients between B-mode ultrasound findings and MRI findings showed medium to large effect sizes (r=0.4-0.8). The Goutallier classification of the infraspinatus muscles was the MRI predictor with the best discriminative power for surgical reparability (area under the curve [AUC], 0.89; 95% confidence interval [CI], 0.81 to 0.98), while the Heckmatt scale for infraspinatus muscles was the most accurate ultrasound predictor (AUC, 0.85; 95% CI, 0.74 to 0.96). No significant differences in AUCs among the MRI and ultrasound predictors were found. Conclusion B-mode ultrasonography was a reliable examination tool and had a similar ability to predict surgical reparability to that of MRI among patients with large-to-massive RCTs.

Ambulance response time is one of the key performance of ambulances services. The objective of this study is to determine the factors associated with delayed ambulance response time in Hospital Universiti Sains Malaysia (HUSM). This was a cross sectional study conducted in Department of Emergency Medicine, Hospital Universiti Sains Malaysia (EDHUSM) between January 2016 to January 2017. A total of 300 ambulance calls were included in our analysis. Data were collected by ambulance paramedic using validated ambulance form. All ambulance forms with missing data were excluded from this study. Of the 300 ambulance calls within the study periods, 254 cases (84.7%) were determined to have delayed ambulance response time. Current ambulance response time is 14 minutes with interquartile range of 5 minutes. Factors which showed significant association delayed ambulance response time include distance from hospital, location, type of emergency and ambulance mechanism. The odd of delayed ambulance response time by every increase in distance unit was 1.59 (95% CI, 1.37 to 1.85). For location type, the odd of delayed ambulance response time for public location as compared to road was 0.13 (95% CI, 0.04 to 0.45). For ambulance mechanism, the odd of delayed ambulance response time for beacon type as compared to siren type was 0.22 (95% CI, 0.01 to 0.69). Further intervention should be initiated based on our findings to improve current ambulance response time.

Objective:To explore whether thioredoin-2 (Trx-2) is involved in the development of cataract and to study the effect of Trx-2 on hydrogen peroxide (H2O2)-induced injury in human lens epithelial cells.Methods:A total of 10 volunteers (removing the lens due totraumatism) and 30 patients received phacoemulsification (age more than 60 years) were selected.The expression of Trx-2 protein in lens epithelial cells from cataract patients and volunteers were detected by the immunohistochemical streptavidin-peroxidase (SP) method.SRA01/04 cells were cultured and were divided into six groups according to different treatment:a control group,H2O2-treated groups at 20,50 or 100 μmol/L,a negative control group (transfected with pCMV6 plasmid plus 100 μmol/L H2O2),and a Trx-2 overexpression group (transfected with pCMV6-Trx-2 plasmid plus 100 μmol/L H2O2).Methyl thiazolyltetrazolium (MTT) assay and flow cytometry was performed to measure the cell viability and apoptosis for SRA01/04 cells,respectively.The activities of superoxide dismutase (SOD) and catalase (CAT),the content of glutathione (GSH) and malondialdehyde (MDA) in human lens epithelial cells were measured via chemical chromatometry.Western blot was used to measure the protein levels of Trx-2,B-cell lymphoma 2 protein (Bcl-2),Bcl-2 associated X protein (Bax) and caspase-3.Results:Compared with the volunteers,the expression of Trx-2 was significantly decreased in lens epithelial cells in patients with cataract (P＜0.05).Compared with the control group,the expression of Trx-2 protein in the 20,50 or 100 μmol/L H2O2 groups was decreased (all P＜0.05).Compared with the control group,the cell survival rates were decreased in the 100 μmol/L H2O2 group and the negative control group (both P＜0.05),along with enhanced apoptotic rates,inhibited cellular SOD activities and CAT activities,reduced GSH contents,augmented MDA contents,down-regulated Trx-2 and Bcl-2 expression and up-regulated Bax and caspase-3 expression (all P＜0.05).Compared with the negative control group,the cell survival rate was increased in the Trx-2 overexpression group (P＜0.05),along with suppressed apoptosis,increased SOD activities and CAT activities,elevated GSH contents,decreased MDA content,up-regulated Trx-2 and Bcl-2 expression and down-regulated Bax and caspase-3 expression (P＜0.05).Conclusion:Trx-2 might be involved in the apoptosis of lens epithelial cells in patients with cataract.The overexpression of Trx-2 obviously attenuated H2O2-induced injury of human lens epithelial cells,which might be associated with the inhibition of H2O2-mediated oxidative stress.

BACKGROUND: Small cell lung cancer (SCLC) is highly malignant and prone to bone marrow metastasis in early stage, but its related reports are limited. This study analyzed the clinical feature, laboratory examination, treatment and prognosis of SCLC patients with bone marrow metastasis. METHODS: The clinical data of 26 SCLC patients with bone marrow metastasis were analyzed retrospectively. Prognostic factors were evaluated. RESULTS: The median age of 26 patients was 57 years and the median time from diagnosis of SCLC to confirmed bone marrow metastases was 8 d. Most patients (96.2%) were accompanied by other organ metastases. The most common laboratory abnormalities were elevated lactate dehydrogenase in 19 cases (73.1%), thrombocytopenia and elevated alkaline phosphatase respectively in 11 cases (42.3%) and anemia in 7 cases (26.9%). Twenty patients had received chemotherapy and the remaining 6 patients had not. Of this group, 16 patients received at least 2 cycles of chemotherapy after the diagnosis of bone marrow metastasis. The median survival time was 15.7 wk (0.1 wk-82.9 wk) after diagnosis of bone marrow metastasis. The survival of patients with chemotherapy was significantly better than that of those without chemotherapy (χ²=33.768, P<0.001). Multivariate analysis showed that no chemotherapy was independent poor prognostic factors (P<0.05). CONCLUSIONS The SCLC patients with bone marrow metastasis have short survival, whereas chemotherapy can extend the survival of patients.
Aged ; Bone Marrow ; pathology ; Bone Marrow Neoplasms ; mortality ; pathology ; secondary ; Female ; Humans ; Lung Neoplasms ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Retrospective Studies ; Small Cell Lung Carcinoma ; pathology

Objective To investigate the roles and mechanisms of siRNA targeted lipocalin 2 (Lcn 2) gene silencing on hypoxia-induced cell apoptosis in retinal ganglion cells (RGC-5).Methods RGC-5 cells were subjected to hypoxic condition for various time duration (0 h,4 h,8 h,12 h,24 h,48 h),and quantitative real-time PCR and Western blot were used to evaluate the expression of Lcn 2 in mRNA and protein levels.Cells were divided into 4 groups:control group,hypoxia group,siNC + hypoxia group,in which cells were transfected with negative control for siRNA,and then subjected to hypoxic condition for 24 h,and siLcn 2 + hypoxia group,in which cells were transfected with Lcn 2 siRNA,and then subjected to hypoxic condition for 24 h).Next,cell apoptotic rate and Caspase-3 activity were detected using ELISA.The fluorescence intensity of reactive oxygen species (ROS) was assayed using DCFH-DA kit,and mitochondrial membrane potential assay kit was used to evaluate the mitochondrial membrane potential.Finally,the expression levels of cleaved-Caspase-3 (c-Caspase-3),cleaved-PARP (c-PARP),Bax,Bcl-2 and cytochrome C (Cyto C) were detected using Western blot.Results In the hypoxia group,Lcn 2 mRNA level at 4 h,8 h,12 h,24 h and 48 h was higher than that at 0 h (all P ＜0.05).Meanwhile,Lcn 2 protein level at 12 h,24 h and 48 h was also higher than that at 0 h (all P ＜ 0.05).Cell apoptotic rate in the hypoxia group (138.33％ ± 13.76％) was significantly higher than that in the control group (99.66％ ± 2.86％) (P ＜ 0.05),and siLcn 2 + hypoxia group (105.02％ ± 8.60％) was lower than siNC + hypoxia group (142.33％ ± 6.54％) (P ＜ 0.05).In addition,compared with the control group,the relative activity of Caspase-3 and the relative expression of c-Caspase-3 and c-PARP in the hypoxia group were all upregulated (all P ＜ 0.05);whereas the relative activity of Caspase-3 and the relative expression of c-Caspase-3 and c-PARP in the siLcn2 + hypoxia group were downregulated compared with the siNC + hypoxia group (all P ＜ 0.05).Moreover,the fluorescence intensity of ROS in the hypoxia group (4.26 0.63) was more enhanced than that in the control group (1.03 ± 0.11) (P ＜ 0.05),and the siLcn2 + hypoxia group (3.10 ± 0.24) was lower than siNC + hypoxia group (4.73 ± 0.26) (P ＜ 0.05).Furthermore,mitochondrial membrane potential,Cyto C expression and the relative ratio of Bax to Bcl-2 in the siLcn2 + hypoxia group was lower than those in the siNC + hypoxia group.Conclusion Lcn 2 Lcn2 gene silencing can inhibit cell apoptosis and ROS production induced by hypoxia,which may be achieved by suppressing mitochondrial apoptosis signaling pathway.

Objective To investigate the dynamics of white blood cell count (WBC),prealbumin(PA),high sensitive C-reactive protein (hs-CRP),and vascular endothelial growth factor (VEGF) in hand-foot-and-mouth disease (HFMD) complicated with or without encephalitis and in vitro inhibitory effects of artesunate on VEGF secretion of mononuclear cells from HFMD patients.Methods Peripheral blood mononuclear cells from healthy control group,HFMD group and HFMD combined with encephalitis group were isolated by Ficoll density gradient centrifugation and treated with different concentrations of artesunate (25,50,100 mg/L).The expression of VEGF in the supernatant was examined by ELISA double antibody sandwich method.The levels of WBC,PA and hs-CRP in the three groups were also detected.Multiple samples were compared by one-way analysis of variance.Multiple comparison was performed by Dunnett T3 test.Correlation of two variables was analyzed by Spearman correlation test.Results There were significant differences in the levels of WBC,hs-CRP,PA and VEGF between the HFMD combined with or without encephalitis group and health control group (F=172.69,366.02,166.32 and 5 941.89,respectively,all P＜0.01).There were significant differences in the levels of VEGF secreted by mononuclear cells treated with three different concentrations of artesunate between the HFMD combined with or without encephalitis groups and health control group (F =194.265 and 4 750.69,respectively,both P＜0.01).Correlation analysis showed that the VEGF level secretion by mononuclear cells from HFMD patients with or without encephalitis were both negatively correlated with different concentrations of artesunate (r=-0.903 and-0.969,both P＜0.01).Conclusions Compared with HFMD without encephalitis,the secretion of VEGF by mononuclear cells,WBC and hs-CRP levels in HFMD complicated with encephalitis group all increase and PA decrease significantly.Artesunate can inhibit the secretion of VEGF by mononuclear cells of patients with hand-foot-and-mouth disease in a dosedependent manner in vitro.

Objective To investigate whether single nucleotide polymorphisms(SNPs)in the ABCC4(rs7320375,rs7329490,rs7986087),FCGR2A(rs1801274)and BLK(rs2254546)region could be susceptibility locus for Kawasaki disease(KD)in children from southern China.Methods This study was performed as a case-control study.The samples,92 individuals with KD and 194 healthy controls from southern China,were collected at Guangzhou Women and Childrens′Medical Center from October,2013 to November,2015,and the SNPs were genotyped by using the Sequenom MassArray system.The genotype distribution and allele frequency of the SNPs were analyzed using chi-square test and Fisher′s exact test.Results The genotype distribution of FCGR2A(rs1801274)in patients with KD were as follows: GG 4.3％(4/92),AG 33.7％(31/92),AA 62％(57/92),correspondingly in healthy controls were GG 48.5％(94/194),AG 41.2％(80/194),AA 10.3％(20/194)respectively,and a significant difference was found between KD patients and controls(x2=98.17,P=0.000).A allele frequency of FCGR2A(rs1801274)in KD patients(78.8％,145/184)was higher than that in controls(30.9％,x2=0.120,P=0.000).The genotype distribution of BLK(rs2254546)in patients with KD were as follows: GG 67.4％(62/92),AG 28.3％(26/92),AA 4.3％(4/92),correspondingly in healthy controls were GG 52.1％(101/194),AG 43.8％(85/194),AA 4.1％(8/194)respectively,significant differences were found between KD patients and controls(x2=6.47,P=0.039).G allele frequency of BLK(rs2254546)in KD patients(81.5％,150/184)was higher than that in controls(74.0％,287/388,x2=1.553,P=0.047).Conclusions For the children in southern China,FCGR2A SNPs(rs1801274)may be associated with the susceptibility to KD,and the A allele may increase the risk of KD.BLK SNPs(rs2254546)is also found to be associated with the susceptibility to KD,and the G allele may increase the risk of KD.