Objective To investigate the diagnostic accuracy and the clinical usefulness of the combination of troponin I (cTnI) and copeptin detected in patients with suspected non-ST elevation myocardial infarction. Methods 176 patients presenting to the emergency departments with chest chocking or chest pain within 6 hours and without ST elevation on a 12-lead electrocardiogram (ECG) were enrolled in this study. The level of copeptin and cTnI was measured. The diagnosis was adjudicated by 2 independent experts.The diagnostic performance of them was assessed using ROC analysis , and the sensitivity and specificity of them were inferred based on the positive rate of two cardiac markers. Results (1)The levels of copeptin and cTnI in NSTEMI patients were markedly higher than other groups (P<0.05).(2)The AUCs of copeptin and cTnI were 0.846 and 0.683, and the 95%CI of two markers were 0.786 ~ 0.906 and 0.577 ~ 0.789, respectively. (3)Using 10.85 pmol/L as cut off value,the sensitivity and specificity of copeptin were 90% and 64%,and the positive predictive value and the negative predictive valueof NSTEMI diagnose were 42.4% and 95.6%,respectively.Using 0.05 ng/mL as cut off value,the sensitivity and specificity of cTnI were 42.5% and 94.1%,the positive predictive value and the negative predictive value were 68%and 84.8% for diagnosis of NSTEMI. (4)The copeptin level over 10.85 pmol/L in combination with cTnI could be used to detect NSTEMI with higher sensitivity than that of copeptin or cTnI alone (95% vs 90% vs 42.5%). The negative predictive value of the combination of copeptin and cTnI was increased , compared to that of copeptin or cTnI alone (97.7% vs 95.6% vs 85.7%). Conclusions Determination of copeptin in addition to cTnI can improves diagnostic performance , especially early after chest pain onset. It seems to allow a rapid and reliable rule out of NSTEMI.

Objective To establish uPA inducible expression system using recombinant retroviral system for the further construction of inducible uPA-SCID animal model .Methods The Inducible expression system need to construct two plasmids:pLNHXO1O2-Alb-GLUC-FMN2A -rtTA and pLNHXO5O6-TRE2-uPA-IRES-ZsGreen respectively. Both plasmids were based on retroviral vector pLNHX , Albumin promoter gene ( Alb) and rtTA gene or uPA gene and ZsGreen were obtained by PCR reaction and inserted into pLNHX .The Gaussia enzyme fluorescent element ( GLUC) was used to monitor rtTA expression in pLNHXO1O2-Alb-GLUC-FMN2A-rtTA, and the ZsGreen for uPA expression monitoring in pLNHXO5O6-TRE2-uPA-IRES-ZsGreen.The correct constructed plasmids were transfected into packaging cell line GP 2-293 to gain recombinant viral particles .NIH/3T3 cells were infected with these viral particles and selected with G 418.Gene expression in the surviving cells was confirmed by the PCR method .Results The recombinant retroviral vectors harbouring target genes were successfully cloned .The rtTA gene in pLNHXO1O2-Alb-GLUC-FMN2A-rtTA was expressed, and uPA can be induced to express in pLNHXO5O6-TRE2-uPA-IRES-ZsGreen by doxycycline (Dox) when the plasmid transfected into the HepG-Tet-on cell.The constructed recombinant two retroviral vectors were transfected into GP 2-293 packaging cells respectively to gain infectious viral particles .Then,NIH/3T3 cells were infected with these viral particles and single-cell clones which stably expressed the transgenes were successfully established .Conclusion This study primarily established uPA inducible expression system , it laid a foundation for the murine model of inducible liver damage , and provided a novel technical platform for further building the liver humanised murine models for viral hepatitis studying .

BACKGROUND:Surgical treatment is commonly used for decompressing the spinal cord in multilevel cervical spondylotic myelopathy, but the optimum anterior cervical reconstructive method has not been determined. OBJECTIVE:To compare and analyze the biomechanical characteristics of four anterior cervical reconstructive techniques in the surgical management of multilevel cervical spondylotic myelopathy utilizing the three-dimensional finite element model. METHODS:A three-dimensional finite element model of an intact C2–7 segment was developed and validated based on healthy adult male CT images. Four additional models were developed from the fusion model, including anterior cervical discectomy and fusion, anterior cervical corpectomy and fusion, anterior cervical hybrid decompression and fusion and anterior cervical discectomy and fusion with Cage alone. Von Mises stresses on the plate and the disc of adjacent levels (C2/3, C6/7) were comparatively analyzed. RESULTS AND CONCLUSION:(1) The C2/3 disc stress in flexion, extension, lateral bending and rotation condition was significantly higher than the C6/7 disc in 4 anterior cervical reconstructive techniques, and the adjacent disc stress (C2/3, C6/7) was lowest in the anterior cervical discectomy and fusion with Cage alone, and highest in the anterior cervical corpectomy and fusion. (2) The stress at the plate-screw interface was highest in the anterior cervical corpectomy and fusion, and lowest in the anterior cervical discectomy and fusion. (3) In conclusion, among the four anterior cervical reconstructive techniques for multilevel cervical spondylotic myelopathy, the anterior cervical discectomy and fusion with Cage alone makes little effect on the adjacent disc stress, which might reduce the incidence of adjacent segment disease after fusion. However, the accompanying risk of the increased incidence of cage subsidence should never be neglected.

ObjectiveTo investigate the clinicopathological characteristics of non-diabetic renal diseases (NDRD) in the patients with diabetes mellitus.MethodsClinicopatholigical data of 202 patients with diabetes mellitus and NDRD identified by renal biopsy from January 1st,2003 to December 31st,2010 were analyzed retrospectively.All the patients were divided into three groups:the young (≤35 years old),the middle-aged (36-59 years old) and the elder (≥60 years old).Clinicopathological characteristics were compared among 3 groups.ResultsIn the young group (n=33),42.4％ of patients presented as chronic glomerulonephritic syndrome,while 36.4％ as IgA nephropathy for pathology.In the middle-aged group(n=136),35.3％ of patients presented as chronic glomerulonephritic syndrome,27.2％ as nephritic syndrome,17.6％ as chronic renal failure,14.7％ as latent glomerulonephritis,and 5.1％ as acute renal failure,while42.6％ as IgA nephropathy for pathology.In the elder group(n=33),30.3％ of patients presented as nephritic syndrome,30.3％ as chronic renal failure,while 27.3％ as membranous nephropathy for pathology.ConclusionsIn clinical manifestation,young patients are mainly chronic glomerulonephritic syndrome,middle-aged patients are diversified,and elder patients are mainly nephritic syndrome andchronicrenalfailure. Inpathology, youngandmiddle-agedpatientsaremainlyIgA nephropathy,and elder patients are mainly membranous nephropathy.

Objective:To describe the expression profiling of microRNAs of patients with non-small cell lung cancer. Methods:We reduced the scope of downregulated microRNAs in patients with non-small lung cancer by high throughput microarray based on quantitative real-time PCR (qRT-PCR). In this process, the downregulated microRNAs of non-small cell lung cancer tissues can be de-tected and compared with adjacent healthy lung tissues with relatively small samples. Different microRNA expression levels in non-small cell lung cancer and adjacent healthy lung tissues were verified in a large sample by RT-PCR. Results:A total of 20 types of microRNAs were significantly downregulated in non-small cell lung cancer tissues compared with adjacent healthy lung tissues. These microRNAs were listed as follows: hsa-miR-1; hsa-miR-22; hsa-miR-27a; hsa-miR-27b; hsa-miR-125a-5p; hsa-miR-125b;hsa-miR-142-5p;hsa-miR-143;hsa-miR-148a;hsa-miR-148b;hsa-miR-370;hsa-miR-373;hsa-miR-381;hsa-miR-489;hsa-miR-519a;hsa-miR-376c;hsa-miR-206;hsa-miR-380-5p;hsa-miR-223-5p;and hsa-miR-520c-3p. Among these microRNAs, 13 types were down-regulated in non-small cell lung cancer tissues as verified by RTPCR. These 13 types of microRNAs were listed as follows:hsa-miR-125a-5p; hsa-miR-143; hsa-miR-519a; hsa-miR-223-5p; hsa-miR-1; hsa-miR-520c-3p; hsa-miR-489; hsa-miR-27a;hsa-miR-373; hsa-miR-125b; hsa-miR-27b; hsa-miR-142-5p; and hsa-miR-206. Conclusion: In non-small lung cancer tissue, several kinds of microRNAs were significantly downregulated. Changes in microRNA expressions were significantly associated with tumori-genesis, progression, or other cancer cell functions in non-small cell lung cancer.

Objective:To explore the status of STAT3 phosphorylation in myeloid-derived suppressor cells (MDSCs) of breast cancer and its function in the immunosuppressive effect of MDSCs on proliferation and cytokine secretion of T cells. Methods:CCD33+cells were isolated from healthy umbilical cord, blood-derived, peripheral blood mononuclear cells and were co-cultured with breast cancer cell line MDA-MB-231 in vitro using Transwell plates to induce MDSCs. The untreated CD33+cells were used as con-trols. Idoxuridine (IDO) suppressor expression and STAT3 phosphorylation were examined using Western blot assay. The proliferation and cytokine secretion of T cells, which were co-cultured with MDSCs, were determined by methyl thiazol tetrazolium assay and en-zyme-linked immunosorbent assay. 1-MT and JSI-124 were used to investigate the function of IDO and pSTAT3 in MDSC-mediated T cell immunosuppression. Results:The protein levels of IDO and pSTAT3 in MDSCs were significantly upregulated. MDSCs obviously suppressed T-cell proliferation, which was reversed by 1-MT or JSI-124 (P<0.05). MDSCs could promote TGF-βand IL-10 secretions, but could also remarkably inhibit IFN-γsecretion (P<0.05). After incubation with 1-MT or JSI-124, the increase in TGF-βand IL-10, as well as the decrease in IFN-γ, was significantly reversed. Conclusion:The upregulated pSTAT3 induced the IDO increase in MDSCs. JSI-124 can block MDSC-mediated immunosuppressive effect on T cells in breast cancer.

Objective:To highlight the developmental process of 3D cell culture technology system, which is more suitable for isolating and identifying lung cancer stem-like cells than 2D cell culture technology system, and to explore the application of 3D cell cultures in the evaluation of proliferation, apoptosis, invasion, and drug resistance of lung cancer. Methods:Cells (104/well) from the human lung adenocarcinoma cell lines A549 and RPMI 1640 were cultured in complete medium containing 10%fetal bovine serum. Cell suspension was cultured in a BME basal medium. A growth curve was drawn after 7 d of culture. The stem-like cell was identified through a mammosphere culture, drug resistance and invasion assay, and flow cytometry. Data of A549 cells cultured in 3D and 2D tra-ditional cell culture technologies were compared. Results: Cells from the 3D cell culture had higher tumor formation rates [(20.75 ± 0.85) d vs. (60.25 ± 1.49) d, P<0.01)] and tumor sphere formation (28.50%± 1.17%vs. 8.67%± 0.80%, P<0.01) than those from the 2D cell culture. Moreover, cells from 3D cell culture were more invasive and resistant to therapy (58.17%± 2.19%vs. 41.70%±5.81%in 48 h, P<0.01;33.27%±5.76%vs. 27.30%±4.25%in 72 h, P<0.01). Phenotype experimental results demonstrated that the CD44 and CD326 cells were double-positive, whereas the CD24 cell was negative. Conclusion:The proportion of stem-like cells in A549 cell line after 3D cell culture significantly increased compared with 2D cell culture. The 3D cell culture can promote the proliferation of lung cancer stem cells.

Objective To investigate effects of melatonin (MT) on high glucose-induced cell proliferation,Toll-like receptor 4 (TLR4) signaling pathway and expressions of inflammatory factor in mouse mesangial cells (SV40).Methods SV40 cells were divided into mannitol control group (30 mmol/L mannitol),normal control group (5 mmol/L glucose),control (5 mmol/L glucose)+ 1000 μmol/LMT group,high glucose group (25 mmogL glucose),high glucose +10,100,1000 μmol/L MT group and high glucose + TLR4 inhibitor (TAK242) group.(1) The cell viability was measured by CCK-8 cytotoxicity kits,and cell proliferation was measured by EdU kits.The expression of TLR4 and the nuclear translocation of nuclear factor-κB (NF-κB p65) were observed by immunofluorescence.(2) Realtime quantitative PCR was used to detect TLR4 mRNA expression.Real-time quantitative PCR and ELISA were used to determine the mRNA and protein secretion levels of the downstream inflammatory factors,such as monocyte chemoattractant-1 (MCP-1),interleukin-1β (IL-1β) and tumor necrosis factor of α (TNF-α);Western blotting was used to detect TLR4 pathway proteins,such as TLR4,myeloid differentiation factor 88 (MyD88),β interferon TIR domain adaptor (Trif),phosphorylated interferon regulatory factor 3 (p-IRF3) and phosphorylated NF-κB inhibitory protein (p-IκB).Results High glucose stimulated mesangial cell proliferation,promoted TLR4 expression and NF-κB p65 transcription activity.Both MT and TAK242 inhibited the above reactions,and the effects of MT was concentration-dependent.Compared with the normal control group,high glucose group had up-regulated expressions of TLR4,MCP-1,IL-1β and TNF-α mRNA (all P ＜ 0.05),but also significantly increased the protein expressions of MyD88,Trif,p-IRF3 and p-IκB (all P ＜ 0.05).Compared with those in the high glucose group,the expression of TLR4 was down-regulated in the high glucose+ 10,100,1000 μmol/L MT group and the high glucose+TAK242 group (all P ＜ 0.05),while the expressions of MyD88,Trif,p-IRF3,p-IκB,MCP-1,IL-1β and TNF-α decreased (all P ＜ 0.05).The effects of MT was concentration-dependent.Conclusions High glucose stimulates the proliferation of SV40,and MT can inhibit the proliferation of mesangial cells and the expressions of inflammatory factors through TLR4 signaling pathway.

Objective:To investigate the incidence of acute kidney injury (AKI) following pancreaticoduodenectomy and related risk factors in elderly patients.Methods:The clinical data of elderly patients who underwent pancreaticoduodenectomy in Henan Provincial People′s Hospital from January 2017 to June 2020 were collected retrospectively. According to the changes of serum creatinine within 48 h or 7 days after operation, the patients were divided into AKI group and non-AKI group. The basic clinical characteristics of the two groups were compared, and the incidence of AKI was calculated. Multivariate logistic regression model was used to analyze the risk factors of postoperative AKI.Results:A total of 322 elderly patients were enrolled, with age of (67.1±5.2) years old (60-85 years old) and 186 males (57.76%). Among 322 elderly patients, there were 41 patients (12.73%) suffering from AKI following pancreaticoduodenectomy. Compared with the non-AKI group, the level of bilirubin in AKI group was higher ( Z=-2.012, P=0.044), and the level of hemoglobin in AKI group was lower ( Z=-2.111, P=0.035). Multivariate logistic regression analysis showed that increased preoperative total bilirubin ( OR=1.003, 95% CI 1.000-1.006, P=0.027) and postoperative exploratory laparotomy ( OR=3.936, 95% CI 1.071-14.460, P=0.039) were the independent influencing factors for AKI after pancreaticoduodenectomy in elderly patients. Conclusions:The incidence of AKI after pancreaticoduodenectomy in elderly patients is 12.73%. Preoperative high bilirubin and postoperative exploratory laparotomy may be the independent risk factors for AKI after pancreaticoduodenectomy in elderly patients.

Objective:To evaluate the efficacy of botulinum toxin A injection therapy with ultrasound-guided for benign masseter hypertrophy.Methods:Twenty cases (40 sides) of masseter hypertrophy were injected with botulinum toxin A, among which 10 cases were taken as experimental group and the other 10 cases as control group. The experimental group had preoperative ultrasound examination to measure the thickness of masseter, and use ultrasound-guided precise injection during the operation, and then ultrasound recheck to measure the postoperative masseter thickness; The other 10 cases of control group adopt traditional 3-point injection method.Results:The masseter thickness of the two groups at different period of times after treatment was decreased ( P<0.001), the most obviously decrease happened 4-12 weeks after injection. The mean reduction was 26.8% at 4 weeks and 28.4% at 12 weeks after injection. Masseter muscle thickness recovered by 22% at 24 weeks and by 20% at 36 weeks. The average follow-up was 6.0±2.4 months with no serious complications occurred. In the control group, there was a partial masseter bulge in one case. The satisfactory rate of the patients in the experimental group was higher than that of the control group. Conclusions:Botulinum toxin A injection therapy with ultrasound-guided for benign masseter hypertrophy is an effective treatment, which is more accurate and effective than the traditional injection method.