Objective To establish U251 cells with inhibited expression of interferon-γ inducible protein 30 (IFI30), and to investigate the effect of IFI30 on cell biological function as well as its underlying mechanism. Methods Three knockdown sequences which target IFI30 were designed online and 3 small interfering RNAs (siRNA) were synthesized. After transfection, the inhibition efficiency was detected by real-time quantitative PCR. The siRNA sequence with the highest inhibition efficiency was selected to create short hairpin RNA (shRNA) plasmids. The recombinant plasmids and packaging plasmids were co-transfected into HEK293T cells to prepare lentivirus. The glioma U251 cells were transfected with lentivirus, and the positive cells were screened by puromycin. CCK-8 assay, 5-ethyl-2'-deoxyuridine (EdU) and colony formation assays were used to analyze cell proliferation; the flow cytometry was used to analyze cell cycle and apoptosis; the Transwell^TM assay was used to detect cell invasion; the wound-healing assay was employed to detect cell migration, and western blot analysis to detect the protein expresison of cyclin D1, B-cell lymphoma factor 2 (Bcl2), epithelial cadherin (E-cadherin), neural cadherin (N-cadherin), signal transducer and activator of transcription 1 (STAT1). Results The sequence which effectively target IFI30 was screened and U251 cell line capable of inhibiting the IFI30 expression was successfully established. When IFI30 expression was knocked down, the proliferation of U251 cells was inhibited, along with increased ratio of cells in the phase G0/G1, the decreased phase S, the increased rate of cell apoptosis. The cell invasion and migration capabilities was also reduced. The decreased expression of cyclin D1, Bcl2 and N-cadherin were observed in U251 cells, and the expression of E-cadherin and the phosphorylation of STAT1 were found increased. Conclusion Knockdown of IFI30 inhibits the proliferation, invasion and migration of human glioma cell U251 and promotes its apoptosis by activating STAT1.
Humans ; Cyclin D1/genetics* ; HEK293 Cells ; Interferon-gamma ; RNA, Small Interfering ; Apoptosis/genetics* ; Cadherins ; Cell Proliferation/genetics* ; Glioma/genetics* ; Proto-Oncogene Proteins c-bcl-2 ; Oxidoreductases Acting on Sulfur Group Donors ; STAT1 Transcription Factor/genetics*

OBJECTIVE: To investigate the growth-inhibitory and pro-apoptotic effects of piroctone olamine (PO) on glioma cells and explore the underlying mechanism. METHODS: Human glioma cell lines U251 and U373 were treated with PO and the changes in cell proliferation were detected using CCK-8 assay and EdU assay. Clone formation assay and flow cytometry were used to examine the changes in clone formation ability and apoptosis of the treated cells. Mitochondrial membrane potential of the cells and morphological changes of the mitochondria were detected using JC-1 staining and a fluorescence probe, respectively. The expressions of mitochondrial fission protein DRP1 and the fusion protein OPA1 were determined with Western blotting. Transcriptome sequencing and differential gene enrichment analysis was performed, and the expression levels of PI3K, AKT and p-AKT in the treated cells were verified using Western blotting. RESULTS: CCK-8 assay showed that PO significantly inhibited the proliferation of U251 and U373 cells in a time- and dose-dependent manner (P < 0.001). EdU test showed that the proliferative activity of PO-treated cells was significantly decreased, and the number of cell colonies also decreased significantly (P < 0.01). PO treatment significantly increased apoptotic rates (P < 0.01), decreased mitochondrial membrane potential and caused obvious changes in mitochondrial morphology of the cells. Pathway enrichment analysis showed that the down-regulated genes were significantly enriched in the PI3K/AKT pathway, which was verified by Western blotting showing significantly down-regulated expression levels of PI3K, AKT and p-AKT in PO-treated cells (P < 0.05). CONCLUSION PO interferes with mitochondrial fusion and fission function through the PI3K/AKT pathway, thereby inhibiting the proliferation and increasing apoptosis of glioma cells.
Humans ; Glioma ; Mitochondrial Dynamics ; Phosphatidylinositol 3-Kinases ; Proto-Oncogene Proteins c-akt

OBJECTIVE: To investigate the effect of lactic acid-induced upregulation of PLEKHA4 expression on biological behaviors of glioma cells and the possible molecular mechanism. METHODS: GEO database and GEPIA2 website were used to analyze the relationship between PLEKHA4 expression level and the pathological grade of glioma. A specific PLEKHA4 siRNA was transfected in glioma U251 and T98G cells, and the changes in cell proliferation ability were assessed by real-time cell analysis technology and Edu experiment. The colony-forming ability of the cells was evaluated using plate cloning assay, and cell cycle changes and cell apoptosis were analyzed with flow cytometry. The mRNA expression of PLEKHA4 was detected by PCR in glioma samples and controls and in glioma cells treated with lactic acid and glucose. Xenograft mice in vivo was used to detect tumor formation in nude mice; Western blotting was used to detect the expressions of cyclinD1, CDK2, Bcl2, β-catenin and phosphorylation of the key proteins in the MAPK signaling pathway. RESULTS: The results of GEO database and online website analysis showed that PLEKHA4 was highly expressed in glioma tissues and was associated with poor prognosis; PLEKHA4 knockdown obviously inhibited the proliferation and attenuated the clone-forming ability of the glioma cells (P < 0.05). Flow cytometry showed that PLEKHA4 knockdown caused cell cycle arrest in G1 phase and promoted apoptosis of the cells (P < 0.01). PLEKHA4 gene mRNA expression was increased in glioma samples and glioma cells after lactate and glucose treatment (P < 0.01). PLEKHA4 knockdown, tumor formation ability of nude mice decreased; PLEKHA4 knockdown obviously lowered the expression of cyclinD1, CDK2, Bcl2 and other functional proteins, inhibited the phosphorylation of ERK and p38 and reduced the expression of β-catenin protein (P < 0.01). CONCLUSION PLEKHA4 knockdown inhibited the proliferation of glioma cells and promoted apoptosis by inhibiting the activation of the MAPK signaling pathway and expression of β-catenin. Lactic acid produced by glycolysis upregulates the expression of PLEKHA4 in glioma cells.
Humans ; Animals ; Mice ; Up-Regulation ; beta Catenin/metabolism* ; Mice, Nude ; Brain Neoplasms/pathology* ; Lactic Acid ; Cell Line, Tumor ; Glioma/pathology* ; Cell Proliferation ; Apoptosis ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; RNA, Messenger/genetics* ; Gene Expression Regulation, Neoplastic

Since December 2019, novel coronavirus pneumonia (NCP) has been reported in Wuhan, Hubei Province, and spreads rapidly to all through Hubei Province and even to the whole country. The virus is 2019 novel coronavirus (2019-nCoV), never been seen previously in human, but all the population is generally susceptible. The virus spreads through many ways and is highly infectious, which brings great difficulties to the prevention and control of NCP. Based on the needs of orthopedic trauma patients for emergency surgery and review of the latest NCP diagnosis and treatment strategy and the latest principles and principles of evidence-based medicine in traumatic orthopedics, the authors put forward this expert consensus to systematically standardize the clinical pathway and protective measures of emergency surgery for orthopedic trauma patients during prevention and control of NCP and provide reference for the emergency surgical treatment of orthopedic trauma patients in hospitals at all levels.

Heavy metal residue in vegetables is a big concern in the whole world.The aim of this work is to explore the effect of multivariable selection on analyzing Cd in Chinese cabbage polluted in lab by collecting the spectra of laser induced breakdown spectroscopy (LIBS) from the samples.At the same time,the actual Cd content in samples was obtained by anodic stripping voltammetry (ASV).The LIBS spectral range in partial least square (PLS) model was screened by standard normal variable transformation (SNV),first derivative (FD),second derivative (SD) and center treatment (CT) for preprocessing spectra and the optimized method was used for the analysis of interval partial least square (iPLS) and synergy interval partial least square (SiPLS).The results indicated that the method of CT was the best as a comparison with PLS,iPLS and SiPLS.And the intervals of wavelength were 214.72-215.82 nm,215.88-216.97 nm and 225.08 -226.35 nm by utilizing the optimized SiPLS.Here the root mean square error of cross validation (RMSECV) between real content and predicted ones was 1.487,the root mean squared error of prediction (RMSEP) was 1.094,the correlation coefficient (R) was 0.9942,and the average relative error (ARE) was 11.60％.The results displayed that LIBS could predict Cd in vegetables by multivariable selection of SiPLS and the accuracy could meet the requirement of rapid and green analysis of Cd in vegetables.

There are many noise signals in original laser induced breakdown spectroscopy (LIBS) spectra.To explore the effect of spectral pretreatment on LIBS information by different filter methods, the LIBS spectra of Pb-polluted cabbage in wavelength range of 400.45-410.98 nm was investigated and preprocessed by adjacent averaging, Savitzky-Golay (S-G) and fast Fourier transformation (FFT).Then partial least square (PLS) model was established for evaluating the spectral treatment effect.The result showed that the root mean square error of prediction (RMSEP) and average relative error of S-G method were 0.26 and 3.7%, suggesting a superior smoothing effect than other methods.Experimental results indicated that an appropriate filtering method could help to improve the spectral quality and raise the precision of model checkout.

Objective To investigate the clinical effects and safety of urethral anastomoses and ureteroscopy urethral realignment in the treatment of urethral straddle injury and catheter placement failure. Methods Ninety patients with urethral straddle injury and catheter placement failure were chosen and divided into A group (45 patients, choosing urethral anastomoses) and B group (45 patients, choosing ureteroscopy urethral realignment). The operation time, intraoperative blood loss, hospital staying time and peri-operation complications in both groups were compared. Results The operation time, intraoperative blood loss, hospital staying time in B group were significantly lower than those in A group: (26.15 ± 10.41) min vs. (71.93 ± 14.50) min, (22.37 ± 7.41) ml vs. (50.70 ± 13.25) ml, (3.22 ± 0.97) d vs. (5.19 ± 1.43) d, P<0.05. After 6 months′follow-up, the clinical indicators in peri-operation period of B group were significantly better than those in A group (P<0.05). The complications incidence in B group was significantly lower than that in A group: 2.22%(1/45) vs. 13.33%(6/45), P <0.05. Conclusions The technology of ureteroscopy urethral realignment in the treatment of urethral straddle injury and catheter placement failure can efficiently shorten the operation time, reduce the degree of trauma and accelerate the rehabilitation process, and it is helpful to reduce the risk of complications in peri-operation period.

Objective:To investigate the characteristics and perioperative management of hemophilia patients with fracture.Methods:Retrospectively, we analyzed 8 patients with hemophilia combined with fracture, who were admittted to our department from 2005 to 2013.Six patients were with hemophilia A and two with hemophilia B;Based on the severity of hemophilia, 2 cases were light, 3 moderate and 3 severe;Based on the location of fracture, 4 cases were femoral neck fractures, 1 femoral intertrochanteric fracture, 1 bilateral distal femur fractures, 1 tibiofibula fracture, and 1 humerus intercondylar fracture. Blood coagulation factor replacement therapy was conducted preoperatively, intraoperatively and postoper-atively, All the patients underwent closed or open reduction and internal fixation or joint replacement. Also, we analyzed the perioperative complications and observed whether the fracture healed.Results:The average age was 33.5 years (14 to 47 years); In 6 cases, fractures occurred at femur, accounting for 75%of all the fractures; Femoral neck fracture was treated by closed reduction and hollow screws fixation;Femoral intertrochanteric fracture, distal femur fracture, and tibiofibula fracture were treated by open reduction and internal fixation with plate;Humerus intercondylar fracture was treated by elbow joint replacement.Intraoperative bleeding was from 50 to 600 mL, an average of 262 mL;Perioperatively, the average use of FⅧ/activated prothrombin complex concentrates ( APCC) was 358 U/kg (125 to 554 U/kg) .Postoperatively, poor wound healing was observed in 2 patients, and the condition improved after symptomatic treatment; In patients with internal fixation, all the fractures united, and the average hea ling time was 14 weeks.No complications such as fixation loosening or rupture occurred after internal fixation.Conclusion:Hemophilia combined with fracture mainly occurred in the young, and the site of fracture was given priority to femur.With perfect preoperative preparation, on the basis of the replace-ment therapy, hemophilia combined with fractures was safe for surgical treatment, and postoperative frac-tures healing wasgood.But the risk of poor wound healing was high.

Objective To analyze the varus deformity of caput humeri and its correlation with fuuctional outcome of unstable proximal humeral fracture managed with locking plate.Methods The study involved 62 patients (24 males and 38 females,at age range of 32-83 years,mean 63.7 years)with proximal humeral fracture who were treated by open reduction and locking plate internal fixation from May 2008 to June 2010 and had full follow-up data.According to Neer classification,there were 11 patients with two parts fractures,41 with three parts fractures,five with four parts fractures and five with dislocation.Their clinical results were evaluated with Constant-Murley scores.ResultsThe mean duration of follow-up was 20.3 months (range,14-37 months),which showed fracture healing in all the patients.There were 11 patients with varus deformity of the caput humeri. The significant difference was found in Constant-Murley score of shoulder function ( P =0.045 ),forward elevation ( P =0.006 ) and abduction (P =0.022) between the varus and non-varus groups.But there was no significant difference in shoulder external rotation( P =0.08 ).The fracture types ( P =0.037 ) and varus deformity of caput humeri ( P =0.006) were closely correlated with the shoulder function between two groups.ConclusionsThe varus deformity of caput humeri mav be predictive factor for poor outcome of the unstable proximal humeral fracture. Restoration of humeral head-shaft alignment and sound reduction should be done as possible during operation.

Objective To investigate the changes in severe burn patients with acute renal failure by using continuous blood purification(CBP). Methods Twelve severe bum patients underwent early CBP were included in this study. The levels of creatinine、blood urea nitrogen and interleukin-6(IL-6) in plasma were determined. The termperature and heart were also observed. Results After the treatment of CBP,the clinical symptoms were improved obviously ( P ＜ 0. 05 ). The levels of creatinine[( 376. 2 ± 24. 5 ) μ mol/L vs ( 145.2 ± 57. 1 ) μmol/L] 、sodium concentration [( 5.3 ± 1.6 ) mmol/L vs ( 4. 0 ± 0. 7 ) mmol/L] and potassium concentration [( 153. 4 ± 13.0 ) mmol/L vs( 140. 1 ±8.6)mmol/L]in plasma were all decreased evidently after the treatment of CBP. There were significant differences ( t = 2. 79,3.06,3. 01, P ＜ 0. 05 ). The levels of IL-6 and APACHE Ⅱ score were ( 264. 31 ± 122.42 ) ng/Land( 15.2 ±5.8) scores before treatment. While after treatment,their levels were(94. 18 ±63. 82) ng/L and(5.4 ±3. 6) scores. The difference was also significant ( t = 4. 17,3. 67, P ＜ 0. 05 ). In this study, 8 patients cured and 4 patients died. Conclusion CBP could eliminate inflammation factors effectively in severe burn patients with acute renal failure. It could afford a useful assist treatment to these patients.