BACKGROUND:Seed cells and scaffold are two key factors for cartilage defects after osteonecrosis of femoral head using tissue-engineered method. OBJECTIVE:To explore the feasibility of Bio-gide col agen membrane combined with dog bone marrow mesenchymal stem cells into chondrocytes. METHODS:Bone marrow mesenchymal stem cells were isolated from beagle dogs by whole bone marrow blood centrifugation method and adherence screening method in vitro and cultured. Morphological changes in cells were observed, and identification was done using cellsurface antigens. Bone marrow mesenchymal stem cells of passage 3 were induced by chondrocyte induction medium to differentiate into chondrocytes (experimental group). cells cultured in normal medium were considered as control group. 3-(4, 5-Dimethylthiazol-2-yl) 2, 5-diphenyl tetrazolium bromide assay was used to measure growth curve of chondrocytes. cells underwent typeⅡcol agen immunohistochemistry and toluidine blue staining. The coculture of bone marrow mesenchymal stem cells at passage 3 and Bio-gide col agen membrane were observed under an inverted phase contrast microscope and scanning electron microscope. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells with high purity and high viability were obtained by whole bone marrow blood centrifugation method and adherence screening method. cells grew wel and had strong amplified ability, and successful y differentiated into chondrocytes. Numerous bone marrow mesenchymal stem cells adhered on the Bio-gide col agen membrane, showing a tendency of multi-layer growth. cells and Bio-gide col agen membrane seem to blend into an integrant part. Cel processes appeared and connected each other and gradual y wrapped the Bio-gide col agen membrane, with the presence of obvious cel matrix secretion. These results suggested that bone marrow mesenchymal stem cells can grow and differentiate into chondrocytes on the Bio-gide col agen membrane.

Objective To explore the clinical outcome of vascularized iliac bone flap combined with tantalum rod implantation for the treatment of osteonecrosis of the femoral head (ONFH).Methods Totally 28 cases (36 hips) of non traumatic avascular necrosis of the femoral head patients from January 2010 to January 2011,which were 15 males and 13 females; a mean age of 40 years (ranged,18-53years),according to ARCO stages:eight hips of stage Ⅱ a,ten hips of stage Ⅱ b,nine hips of stage Ⅲ a,nine hips of stage Ⅲ b,adopt to vascularized iliac grafting combined with tantalum rod Implantation for osteonecrosis of the femoral head.The Harris hip score and VAS score of pre-and post-operation was recorded to evaluate the clinical outcome,and to compare observe and analyze the change of postoperative of patients by used three-dimensional gait.Results All 28 cases (36 hips) were followed up 6 ～20 months,averaged 12 months.The results of Harris hip score and VAS score of the 6 months postoperatively and last follow-up were significantly higher than preoperative ones (P ＜0.05).And the Harris hip score of the last follow-up was also significantly higher than the one of 6 months postoperatively (P ＜ 0.05).However,there was no significant different in that of VAS score (P ＞0.05).There were 13 cases in excellent,fourteen in good and 4 in fair with an excellent and good rate of 89.8％,which shows that the extension of time and changes in patients gait tends to normal by three-dimensional gait analysis.Conclusion Vascularized iliac bone flap transplantation combined with tantalum screw was an effective method with high clinical success rate for treatment of younger patients with early-mid stage ONFH.It provided good blood supply and enough mechanical support to reduce the progress of femoral head collapse.

BACKGROUND:There are a variety of treatments for femoral head necrosis,but their efficacy is not confirmed and unified.How to improve the differentiation ability of osteoblasts in the femoral head and improve the biomechanical support after the repair of the femoral head is an urgent problem to be solved.OBJECTIVE:To explore the clinical outcome of stem cells combined with vascularized iliac bone flap and tantalum rod implantation for the treatment of osteonecrosis of the femoral head (ONFH).METHODS:Totally 28 cases (36 hips) of non-traumatic ONFH admitted at the Zhongshan Hospital of Dalian University from January 2010 to January 2011 were enrolled.Bone marrow samples were extracted from each patient to isolate bone marrow stromal stem cells which were cultured in vitro for 2 weeks.Tantalum rod implantation with vascularized iliac bone graft was conducted to restore the femoral head shape,and then,prepared stem cell suspension were injected into the iliac bone flap and into the subchondral space of the femoral head.RESULTS AND CONCLUSION:All the 28 cases (36 hips) were followed up for 6-20 months (average 12 months),and their Harris hip scores and visual analogue scale scores at postoperative 6 and 12 months were significantly higher than the baseline (P ＜ 0.05).The Harris hip score at postoperative 12 months was significantly higher than that at postoperative 6 months (P ＜ 0.05),but there was no significant difference in the visual analogue scale scores at 6 and 12 months postoperatively (P ＞ 0.05).At the end of 12-month follow-up,clinical outcomes were excellent in 13 hips,good in 15 hips,fair in 4 hips,and poor in 4 hips,with an excellent and good rate of 90％.These findings indicate that autologous bone marrow stromal stem cell transplantation with vascularized iliac bone flap and tantalum rob implantation is an effective method with high clinical success rate for the treatment of ONFH.

Objective:To investigate the clinical value of peripheral monocyte and neutrophil count in predicting the response of patients with metastatic non-small cell lung cancer (mNSCLC) to immunosuppressive checkpoint inhibitors (ICI).Methods:The clinical data of 34 adult mNSCLC patients who received nafulizumab or pabolizumab in Danzhou People's Hospital of Hainan Province from January 2017 to March 2019 were retrospectively analyzed. The correlation of the demographic characteristics, clinical data, hematological examination results in the first two weeks before the treatment and two weeks after ICI treatment with prognosis was recorded and observed.Results:The baseline mean monocyte count [(0.52±0.09)×10 9/L vs. (0.60±0.12)×10 9/L] and neutrophil count [(4.27±0.87)×10 9/L vs.(5.39±1.02)×10 9/L] of patients with ICI reaction were lower than those of patients without ICI reaction, and the differences were statistically different ( t = -2.572, -2.727, all P < 0.05). However, there was a negative correlation between the monocyte count of the patients who responded to ICI and the reaction time ( r = -0.507, P < 0.05). The median reaction time in patients with monocyte count >0.70×10 9/L was shorter than that in patients with monocyte count ≤0.70×10 9/L (8 weeks vs. 12 weeks, χ2=4.162, P = 0.041). There was no correlation between monocyte count and time of reaction duration, progression of free survival (PFS) and overall survival (OS) ( r = -0.214, 0.182, 0.232, all P > 0.05). The decrease rate of neutrophil count in response group was higher than that in non-response group (22% vs. 2%, P < 0.05). After the first administration, cutoff value of neutrophil count was 4.2×10 9/L; the response rate of patients with neutrophil count ≤ 4.2×10 9/L was higher than that of patients with neutrophil count > 4.2×10 9/L [86.7% (13/15) vs. 36.8% (7/19), χ2=6.657, P < 0.05]. Conclusion:Peripheral blood monocyte and neutrophil count can predict the response to ICI therapy in patients with mNSCLC.

OBJECTIVETo explore the mechanism of hepatitis B virus X protein (HBx)-mediated inhibition of CYP2El expression and its significance in hepatocellular carcinoma (HCC) metastasis.

METHODSA deletion series and mutagenesis series of human CYP2E1 promoter sequence was co-transfected with the HBx expression vector pCMV-2B-FLAG-X into the human HepG2 hepatoma cell line. Reverse transcription-PCR and real-time PCR were used to evaluate the effects of HBx on CYP2E1 promoter activity. The luciferase reporter gene assay was used to identify the HBx-responsive region in the CYP2E1 promoter. Electrophoretic mobility shift assay was used to detect the protein complexes binding to nucleic acids in the CYP2E1 promoter. Martrigel invasion assay was used to examine effects of HBx-inhibited CYP2E1 on invasiveness.

RESULTSAnalysis of the deletion series and mutagenesis series led to identification of two regions of sequence in the CYP2E1 promoter that are important in HBx-mediated modulation of CYP2E1 activity in HepG2 cells (F = 112.24, P = 0.001). Both HNF4alpha and SREBP-1, which directly interact with CYP2E1 promoter sequences, were implicated in the mechanism of HBx-mediated modulation of CYP2E1 promoter activity. In addition, PI3K and JNK pathways were involved in the HBx-mediated modulation (t = 8.56, P = 0.0012 and t = 10.25, P = 0.0009 respectively). HBx-mediated repression of constitutive CYP2E1 led to increased invasiveness.

CONCLUSIONHBx-mediated inhibition of CYP2E1 expression may promote HCC by increasing tumor progression and invasiveness through modulation of the PI3K and JNK signaling pathways.

OBJECTIVETo explore the effects of c-met-siRNA on the growth and invasion of hepatocellular carcinoma MHCC97-H cells by pSuppressorRetro/c-met-siRNA recombinant plasmid transfection.

METHODSRecombinant plasmid transfection to Phoenix A cells was constructed using the lipofectin method and then the retrovirals containing c-met-siRNA were used to infect target cells MHCC97-H. In vitro, c-met expression was tested by Western blot. Cell proliferation, motility and invasiveness were studied using MTT, cell migration assay, and cell invasion assay, respectively.

RESULTSThe expression of c-met decreased significantly in MHCC97-H cells, and the most effective site of the target sequence was at 537. The growth, motility and invasiveness of MHCC97-H cells were inhibited.

CONCLUSIONThe results indicate that c-met-siRNA can down-regulate the expression of c-met and inhibit hepatocellular carcinoma cell proliferation, motility and invasiveness.

Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Plasmids ; Proto-Oncogene Proteins c-met ; genetics ; metabolism ; RNA, Small Interfering

OBJECTIVE: To investigate the effect and mechanism of bactericidal/permeability-increasing protein (BPI) on sepsis induced by intra-abdominal infection in rats. METHODS: Cecal ligation and puncture (CLP) was made on 20 rats with sepsis induced by intra-abdominal infection. BPI or equal volume of physiological saline (PS) was intra-abdominally given immediately and 12 h after CLP, respectively (2.5 mg/kg of BPI each time). Plasma endotoxin levels were determined with limulus amebocyte chromogenic assay. RESULTS: (1) The survival time in BPI group was significantly higher than that in PS group. (2) The values of the mean arterial pressure (MAP), the left ventricular systolic pressure (LVSP), the isovolumic ventricular pressure (IP), and the maximal change of left intraventricular pressure (+/-dp/dtmax) in BPI group, although decreasing, were markedly higher than those in PS group. (3) Plasma glutamic-pyruvic transaminase (GPT) and urea nitrogen levels in BPI group, though increasing, were obviously lower than those in PS group. (4) There was no significant change of plasma endotoxin levels in BPI group, while plasma endotoxin levels markedly increased in PS group. CONCLUSIONS: BPI has obvious protective effect on sepsis induced by intra-abdominal infection, which might be related to its neutralization of endotoxin.

OBJECTIVE: To explore the effects of c-Met-siRNA on the proliferation, movement and invasion of laryngeal carcinoma Hep-2 cells in vitro. METHOD: Firstly, the pSilencer 2.0/c-Met-shRNA recombinant plasmid was transfected into laryngeal carcinoma Hep-2 cells with transfecting agent of cationic liposome Lipofectamine 2000. Secondly,the transfection efficacy was tested by RT-PCR and Western-Blot, then the most inhibitive c-Met-siRNA sequence was elected. Cell proliferation, movement and invasion were detected with MTT, cell migration assay and cell invasion assay, respectively. RESULT: After the transfection of pSilencer 2.0/c-Met-shRNA recombinant plasmid into laryngeal carcinoma Hep-2 cells, the expression of mRNA and protein of c-Met decreased significantly in Hep-2 cells, and ability of the proliferation, movement and invasion of laryngeal carcinoma Hep-2 cells were also inhibited. CONCLUSION The results indicated that c-Met-siRNA can down-regulated the expression of c-Met and markedly inhibited laryngeal carcinoma Hep-2 cell proliferation, movement and invasion. It may have the potential as a therapeutic modality to treat human laryngeal carcinoma.
Apoptosis ; genetics ; Carcinoma, Squamous Cell ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Laryngeal Neoplasms ; genetics ; pathology ; Liposomes ; Proto-Oncogene Proteins c-met ; genetics ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transfection

A novel coronavirus pneumonia (NCP) epidemic has occurred in Wuhan, Hubei Province since December 2019, caused by a novel coronavirus (2019-nCoV) never been seen previously in human. China has imposed the strictest quarantine and closed management measures in history to control the spreading of the disease. However, severe trauma can still occur in the NCP patients. In order to standardize the emergency treatment and the infection prevention and control of severe trauma patients with hidden infection, suspected or confirmed infection of 2019-nCoV, Trauma Surgery Branch of Chinese Medical Doctors' Association organized this expert consensus. The consensus illustrated the classification of the NCP patients, severe trauma patients in need of emergency surgery, emergency surgery type, hierarchical protection for medical personnel and treatment places. Meanwhile, the consensus standardized the screening, injury severity evaluation, emergency surgical treatment strategy and postoperative management strategy of severe trauma patients during the epidemic period of NCP, providing a basis for the clinical treatment of such kind of patients.