Objective To study the inhibition of tumor necrosis factor-alpha(TNF-α)cytokine expression of BV-2 cells induced by hypoxia-reoxygenation injury by siRNA targeting TLR4 gene via the RNAi mechanisms.Method BV-2 mouse microglial cell line was cultured in six-well plates and randomly divided into group N(nor-mal group),group H(hypoxia-reoxygenation),group T(hypoxia-reoxygenation+TLR4-siRNA transfected group),group C(hypoxia-reoxygenation+pEGFP-H1/control-siRNA transfected group)and group B(hypox-ia-reoxygenation+pEGFP-H1 transfected group).Group H,group T,group C and group B were cultured in hy-poxia condition for 3 h followed by reoxygenation for 24 h.The plasma was transfected into BV-2 cells mediated by lipofectamine 2000.The efficiency of transfection were detected by flow cytometry to observe the expression of EGFP.RT-PCR method was used to detect the level of mRNA of TLR4 or NF-кB p65.Westem blot methed was used to test the expression of TLR4 protein.and ELISA was used to test the level of TNF-α in the supernatants.Analysis of variances was used for statistical analysis.Results The expression of EGFP gene waa;(67.58±7.16)% after transfection by flow cytometry analysis.Compared to group N,the TLR4 mRNA,NF-кB p65 mR-NA,TLR4 protein level and the TNF-α quantity in group H,group T,group C and group B increased after the hy-poxia-reoxygenation treatment(P＜0.01).While the expression of the TLR4 mRNA,NF-кB p65 mRNA,TLR4 protein level and the TNF-α quantity in the group T down-regulated compared to group H,group C and group B(P＜0.01).And there were no changes in group C,group B and group H about observation index(P＞0.05).Conclusions The siRNA targeting TLR4 mRNA could inhibit the inflammatory reaction released by BV-2 cells in-duced by hypoxia-reoxygenation stimulation.

Objective To analyse the clinical characteristics,therapeutic modalities and prognosis of pulmonary large cell neuroendocrine carcinoma(LCNEC). Methods The clinical data of 14 patients with pulmonary LCNEC confirmed by surgery were retrospectively reviewed. Results No case was correctely diagnosed before surgery.The immunohistochemical amalysis of the specimens revealed the characters of endocrine carcinoma.Twelve cases received adjuvant chemotherapy of platinum agents,but recurrence or metastasis was found in 8 of them several months after surgery.The median survival time was 19 months.The 1-year,2-year and 3-year survival rate were 85.7%(12/14),21.4%(3/14) and not more than 14.3%(2/14),respectively.The statistical analysis showed that the stage,lymph node metastasis and postoperative adjuvant chemotherapy may have impacts on the prognosis of pulmonary LCNEC. Conclusion Pulmonary LCNEC is a carcinoma with poor prognosis,high tendency of invasion and metastasis.The stage of disease,lymph node metastasis,and adjuvant chemotherapy may be related to the prognosis.

Objective To investigate the effects of enteral nutrition support with different doses of glutamine on nutritional status and gastrointestinal function in elderly critically ill patients.Methods Totally 90 critically ill patients were randomly divided into three group:the control group without glutamine (group A),enteral nutrition with low glutamine [0.3 g/(kg· d)] (group B),and high glutamine [0.6 g/(kg· d)] groups (group C).Fasting blood samples were collected on days 1,7,and 14 after the initiation of study,and then the indicators including serum diamine oxidase,glutamine,transferin (TF),prealbumin (PA),hemoglobin (HGB),albunin,and nitrogen balance were determined.In addition,the gastrointestinal complications including bloating,diarrhea,and gastric retention were recorded.Results On the 7th and 14th days,the serum glutamine levels in groups B and C were (1 886.8 ±420.5),(2 228.6 ±440.2) μg/L;(1 889.6 ± 436.1),(2 174.3 ± 440.8) μg/L,respectively,which were significantly higher than those in group A [(1 612.0 ±493.5),(1 869.7 ±559.8) μg/L] (P =0.027,P =0.008).The serum diamine oxidase levels were also significantly lower in group B [(2 310 ± 1 271),(1 602.5 ± 1 137.9) U/L] and group C [(2 076.3 ±567),(1 586.3 ±530.9) U/L] than in group A [(3 250 ±923),(2 476 ±862) U/L] (P =0.000,P =0.000).The serum TF and PA levels were significantly increased on the 7th day (P =0.023,P =0.047),and the HGB was significantly higher on 14th day (P =0.003),The number of patients who had undergone successful implementation of enteral nutrition were 4 and 3 in groups B and C,and the number of patients who had achieved the target volume of enterral nutrition was 25 and 27 in groups B and C,which were significantly larger than those in group A (12 and 13,respectively) (P =0.008,P =0.000).The gastrointestinal complication was also significantly decreased after the administration of enterral nutrition with glutamine.Conclusions Enteral nutrition with glutamine can improve intestinal mueosa barrier and nutritional status in elderly critically ill patients.Thus,it helps to reduce gastrointestinal complications and transition time of total enteral nutrition.

Adult ; Bone Nails ; Female ; Femoral Fractures ; surgery ; Femoral Neck Fractures ; surgery ; Femur ; injuries ; surgery ; Fracture Fixation, Intramedullary ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo explore the regulatory effect of Suifukang (SFK), a compound Chinese patent drug on the change of free radical in tissue of spinal cord and blood caused by experimental spinal injury.

METHODSSeventy male adult SD rats were randomly divided into 4 groups, the SFK group (n = 20), the hormone group (n = 20), the control group (n = 20) and the normal group (n = 10). Rats in the former three groups were made into the right hemitransected spinal cord (T12) injury. SFK were given to the rats in the SFK group 3 days before and 2 hrs after modeling via gastrogavage, saline was given to rats in the control group at the same time. As for the rats in the hormone group, methyl-prednisolone 30 mg/kg was given once immediately by peritoneal injection. The rats were sacrificed in two batches (10 of each group in each batch) at 8 hrs and 24 hrs after modeling to obtain the serum and injured spinal cord tissue for determining superoxide dismutase (SOD) activity and malondialdehyde (MDA) content.

RESULTS(1) As compared with the normal group, SOD activity in serum and spinal tissue of the control group was lower significantly both at 8 hrs and 24 hrs after modeling, but the changes of SOD activity in the SFK and the hormone group were insignificant; (2) Content of MDA in serum and spinal tissue of rats elevated after modeling, which in the control group at 8 hrs after modeling were higher than that in the normal group, also higher than that in the SFK group and the hormone group; but at 24 hrs after modeling, it lowered significantly in both treated groups. The changes of MDA content in serum were similar to those in the spinal tissue.

CONCLUSIONSFK could effectively eliminate the excessive free radical in serum and injured spinal tissue, and raise the capability of antioxidation of organism.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Free Radical Scavengers ; pharmacology ; therapeutic use ; Male ; Malondialdehyde ; metabolism ; Neuroprotective Agents ; pharmacology ; therapeutic use ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; drug therapy ; metabolism ; Superoxide Dismutase ; metabolism

OBJECTIVETo compare the effects and complications of subinguinal microscopic ligation, laparoscopic transperitoneal varicocelectomy, laparoscopic retroperitoneal varicocelectomy, open retroperitoneal high ligation, and interventional embolotherapy in the treatment of varicocele.

METHODSWe conducted a retrospective study that included 632 varicocele patients treated by subinguinal microscopic ligation (group A, n = 79), laparoscopic transperitoneal varicocelectomy (group B, n = 120), laparoscopic retroperitoneal varicocelectomy (group C, n =137), open retroperitoneal high ligation (group D, n = 283), and interventional embolotherapy (group E, n = 13). We compared the baseline and 3-month postoperative semen parameters, postoperative complications, and pregnancy rate among the five groups of patients.

RESULTSThe operation time was longer in groups A ([2.02 ± 1.25] h) and E ([2.17 ± 1.02] h) than in the other three groups, while the postoperative hospital stay was the shortest in group E ([1.1 ± 0.1] d). Intestinal injury or incision bleeding occurred intraoperatively in 2 cases in group B and 1 case in group E. Postoperative scrotal edema developed in 3.7, 17, 10, and 19% of the patients in groups A, B, C, and D, respectively, but not in group E. The rate of 1-year recurrence was the lowest in group A (1.6%) and highest in group E (22%). Sperm concentration and the percentages of progressively motile sperm and morphologically normal sperm were improved postoperatively in all the patients (P < 0. 05), but there were no statistically significant differences among the five groups either in the above three parameters or in the postoperative pregnancy rate (P > 0. 05).

CONCLUSIONIn the surgical treatment of varicocele, laparoscopic retroperitoneal approach involves short operation time and few complications, subinguinal microscopic ligation has the advantages of little injury, rapid recovery, and few complications but requires specialized microsurgical techniques, and interventional embolotherapy leaves no incision scar and needs only local anesthesia and 1-day postoperative hospital stay, which is uitable for those with a contraindication to anesthesia.

Embolization, Therapeutic ; adverse effects ; methods ; Female ; Humans ; Laparoscopy ; adverse effects ; methods ; Length of Stay ; Ligation ; adverse effects ; methods ; Male ; Operative Time ; Postoperative Complications ; Postoperative Hemorrhage ; Pregnancy ; Pregnancy Rate ; Recurrence ; Retroperitoneal Space ; Retrospective Studies ; Sperm Count ; Urogenital Surgical Procedures ; adverse effects ; Varicocele ; surgery ; Vascular Surgical Procedures ; adverse effects ; methods

OBJECTIVETo compare the biomechanics of three internal fixations for distal femoral fractures,and to choose suitable clinical internal fixation according to experimental date.

METHODSThe fracture models of femoral condydle were made on 12 fresh cadaver femurs (C1 type of AO classification). The fractures were fixed by supracondylar intramedullary interlocking nails (A), L-shaped condyle plates (B) and compressed plates (C) respectively. The straining, displacement, rigidity and intensity of femoral supracondyla of the three internal fixations of distal femoral fractures were observed and analyzed by biomechanics. These three different internal fixation methods were compared by experimental stress analysis. These results were further confirmed by theoretical analysis using finite element method (FFM).

RESULTSUnder the same load condition, supracondylar intramedullary interlocking nail fixation was the best on the intensity and rigidity,while for compressed plate and L-shaped condyle plate, the femoral condyle intensity and rigidity were worse (P < 0.05). There were distinct differences in stress and displacement among three methods (P < 0.01).

CONCLUSIONSupracondylar intermedullary interlocking nail is the best method among the three internal fixation methods. It has advantage such as screw location, solid fixation and less complication. It is a good internal fixation method for distal femoral fracture.

Adult ; Biomechanical Phenomena ; Femoral Fractures ; physiopathology ; surgery ; Finite Element Analysis ; Fracture Fixation, Internal ; methods ; Humans ; Male

OBJECTIVETo identify radiographic predictors of residual low back pain (LBP) after laminectomy for lumbar canal stenosis (LCS).

METHODSClinical results and radiographic findings in 69 patients who underwent single level laminectomy for LCS were retrospectively reviewed. Patients who had an improvement in LBP scores evaluated by Japanese Orthopaedic Association (JOA) scoring system during the follow-up periods were classified as the recovery group, and others were classified as the non-recovery group. Patients'clinical data and radiographic parameters like lordosis angle, range of motion and intervertebral rotational angle were analyzed using binary logistic regression analysis to detect factors significantly related with the occurrence of residual LBP.

RESULTSThe average preoperative JOA score of 14.8+/-5.05 improved to 21.59+/-5.51 at the final follow-up. Binary logistic regression analysis revealed that significant predictors of residual LBP were preoperative lumbar lordosis angle and range of motion.

CONCLUSIONSOur results suggest that patients with flat back and limited lumbar mobility before surgery tend to have poor results in terms of LBP. Therefore, these sagittal radiographic parameters should be taken into account when choosing laminectomy as the surgical option for LCS.

Aged ; Female ; Humans ; Laminectomy ; Low Back Pain ; diagnostic imaging ; Lumbosacral Region ; Male ; Prognosis ; Radiography ; Retrospective Studies ; Spinal Stenosis ; surgery ; Spine ; diagnostic imaging ; Treatment Outcome

E2 gene of classical swine fever virus (CSFV) was cloned into secretory pPIC9K Pichia pastoris expression vector. After being linearized by digestion, the vector was transformed into Pichia pastoris by electroporation to integrate with the genome, the transformants with high copies were screened by G418 and were induced to express with methonal. The results of SDS-PAGE and Western blot demonstrated that the supernatant of the induced P. pastoris culture contained protein E2. The results of the study on the immunological activity indicated that the protein E2 expressed in P. pastoris can elicit animal bodies to produce antibodies against protein E2.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Classical swine fever virus ; genetics ; immunology ; Cloning, Molecular ; Gene Expression ; Pichia ; Rabbits ; Swine ; Viral Envelope Proteins ; genetics ; immunology