Objective To research Dengue virus type 2 (DENV-2) to human umbilical vein endothelial cells (HUVEC) permeability. Methods The titration of DENV-2 was detected and HUVEC infection DENV-2 layer of cells. At different time points after infection (30 min, 1 h, 3 h, 6 h, 12 h, 30 h,36 h, 24 h, 42 h, 48 h,72 h), the permeability in HUVEC were detected after Dengue virus infection. The virus load in HUVEC was detected by quantitative real-time PCR. And ultrastructure in HUVEC was observed by electron microscope. Results The permeability in HUVEC was changed after Dengue virus infection by time lasting. The most permeability in HUVEC was changed after Dengue virus infection at 30 min and 42 h. And at the same time the virus load were most value in all of time. The results of the cell membrane were changed by electron microscope. The deciduous cell microvillus and dissd endochylema were founded. And some of nuclear membrane blank was wided by Dengue virus. Even the leakage was founded in cell nucleus. The other change included that disappearance mitochondrial and vacuolization crista, elder pith change mitochondria. In addition, the complete of cell membrane were dissolved. Conclusion The permeability of HUVEC was changed by DENV-2. To explore the pathogenesis of Dengue virus provide certain theoretical basis.

Objective:To silence ERCC2 (excision repair cross-complementing rodent repair deficiency,complementatin group 2,ERCC2) expression in esophageal cancer KYSE150 cells by small interfering RNA (siRNA) and observe the altered sensitivity of KYSE150 cells to paclitaxel (PTX) and elucidate the mechanism underlying the reversion of the PTX resistance of KYSE150 cells. Methods:ERCC2-targeted siRNA was synthesized in vitro and transiently transfected into ERCC2 overexpressing KYSE150 cells via Lipofectamine mediation. The mRNA and protein expression levels of ERCC2 were determined by using RT-PCR and FCM method, respectively. The sensitivity of KYSE150 cells to PTX was measured by MTT assay before and after siRNA transfection. Results:RT-PCR results suggested that the specific bands of ERCC2 mRNA were not detected in si-ERCC2 group at 24, 48 and 72 h post transfection. FCM results indicated that the expression levels of ERCC2 protein gradually decreased by 31.2%, 51.6% and 60.0% at 24, 48 and 72 h respectively(P<0.01). The IC_(50) value of PTX for ERCC2-silencing KYSE150 cells was (6.32±0.87) μg/mL, lower than that for control cells (P<0.01). Conclusion:siRNA successfully silenced the expressions of target gene ERCC2 at both the transcription and translation levels. Silencing ERCC2 expression partly reversed the resistance of KYSE150 cells to PTX.

BACKGROUND:Twin block appliance has been shown to effectively promote mandibular bone growth of Class Ⅱ malocclusion patients and improve lateral appearance of mandibular retrusion.The other improvement of Twin block appliance on hard-soft tissues remains unclear.OBJECTIVE:To assess the hard-soft tissue changes following Twinblock appliance treatment.METHODS:Atotal of 48 adolescent patients,30 males and 18 females,aged 11.6 years (range 9.58-12.37) with Class Ⅱ malocclusion were selected from Department of Orthodontics,Affiliated Stomatology Hospital of Hebei Medical University.Of them,27 were treated with Twinblock appliance as the treatment group,and 21 adolescent patients without treatments were selected as the control group.Cephalonetric analysis was used to compare the changes before and after treatment.RESULTS AND CONCLUSLON:Comparative analysis showed that Twinblock could effectively adjust or improve the effect of the natural growth,reduced upper lip convexity and mentolabial furrow angle.Moreover,Twinblock inhibited facial vertical distance reduction and facial convexity increase to decrease the potential differences between maxillary and mandibular bones in sagittal plance.Results show that Twinblock could effectively improve the profile of soft-tissues of Class Ⅱ malocclusion cases,

Aim To study mRNA and protein expression of eNOS in pulmonary tissue of chronic hypoxic pulmonary hypertension(HPH)rats and chronic hypoxic rats treated with novel KATP opener iptakalim.Methods sixty Sprague-Dawley(SD)male rats were randomly divided into control group, hypoxic group, low dose iptakalim group(0.75 mg·kg~(-1)·d~(-1)), and high dose iptakalim group(1.5 mg·kg~(-1)·d~(-1)).Except the first group, the other three groups were put into hypoxic and normobaric chamber (10%±0.5% O_2,8 h/day and 6 day/week) to establish chronic hypoxic model. After four weeks, the mean pulmonary arterial pressure(mPAP), RV/(LV+S)and the plasma concentration of NO were measured. RT-PCR was performed to analyze the mRNA expression of eNOS in pulmonary tissue. Western blot was performed to analyze the protein expression of eNOS, iNOS in pulmonary tissue. Results ① The level of mPAP and RV/ ( LV + S) were significantly higher in the hypoxic group than those in control group ( P < 0. 05 ) , Low dose iptakalim groupandhighdoseiptakalimgroupdecreased the level of mPAP and RV/( LV + S) significantly (P <0. 05). ② The level of NO was significantly lower in the hypoxic group than those in control group (P<0. 05). Low dose iptakalim group and high dose iptakalim group increased the level of NO significantly (P < 0. 05 ). ③ The mRNA and protein expression of eNOS in the hypoxic group were significantly lower than those in the control group (P < 0. 05 ). Low dose iptakalim group and high dose iptakalim group increased the expression of eNOS significantly ( P < 0. 05). High dose iptakalim group was more significant. Conclusion Pulmonary vascular endothelial dysfunction is induced by chronic hypoxia,and the level of NO, the mRNA and protein expression of eNOS are decreased. Iptakalim can improve the vascular endothelial dysfunction, increase the expression of eNOS and the level of NO and reverse hypoxic pulmonary hypertension.

Objective To investigate the CT radiological features of pancreatic cystadenomas and cystadenocarcinomas. Methods The CT scans from 39 patients with pathologically proven cystic pancreatic tumors (21 cases of serous cystadenomas, 12 cases of mucinous cystadenomas, and 6 cases of mucinous cystadenocarcinomas) were retrospectively analyzed. Tumor location, the number of cyst (polycystic or not) , diameter of the largest cyst, features of the cyst wall, partition within cyst, border of tumor, and the relationship between tumors and pancreatic duct were recorded. Results In 21 patients with serous cystadenomas, tumors were located at the pancreatic head and neck areas in 17 cases, at the pancreatic body and tail areas in 5 cases, and 1 case was multiple. All 21 cases were polycystic; the median diameter of the largest cyst was 1.77 cm; calcification was seen in cyst wall or partition in 4 cases and soft tissue was seen in 7 cases; mild pancreatic duct dilatation was found in 10 cases. In 12 patients with mucinous cystadenomas, tumors were located at the pancreatic head and neck areas in 6 cases, at the pancreatic body and tail areas in6 cases; 4 cases were polycystic; the median diameter of the largest cyst was 4.88cm; calcification was seen in 1 case and soft tissue was seen in 6 cases; pancreatic duct dilatation was found in 2 cases and mild duct dilatation was found in 3 cases. In 6 patients with mucinous cystadenocarcinomas, tumors were located at the pancreatic body and tail areas in 5 cases, tumor was located at the pancreatic head and neck areas in 1 case; 4 cases were polycystic; the median diameter of the largest cyst was 5.09 cm; calcification was seen in 1 case and soft tissue was seen in 5 cases; duct dilatation was found in 1 case. In all cases, there was no pancreatic duct communication. After enhancement, the soft tissue and partition of lesion was enhanced to gome extent. Conclusions There are CT radiological features of pancreatic cystadenomas and cystadenocarcinomas. However, there are still some atypical CT appearances that may challenge the diagnosis.

OBJECTIVETo evaluate the clinical results of using end plate rings in preventing subsidence of titanium cage in anterior cervical corpectomy and fusion (ACCF) surgery.

METHODSThe clinical data of 71 patients with cervical spondylotic myelopathy underwent ACCF in single segment from February 2008 to February 2011 were retrospectively analyzed. There were 38 males and 33 females, aged from 39 to 74 years old with a mean of 53.8 years. Thirty-three were used end plate rings and thirty-eight were not used (end plate rings group and no end plate ring group, respectively). The Japanese Orthopaedic Association (JOA) score, Odom's scale, imaging data were used to evaluate the clinical effects. Imaging data including Cobb angle of fusion segment, intervertebral height of anterior border (Da) and posterior border (Dp), the mean intervertebral height (Dm).

RESULTSAll patients were followed up from 13 to 34 months with an average of 19.5 months. Between two groups, there was no significant difference in Cobb angle of fusion segment and the mean intervertebral height (Dm) before surgery and one week after surgery. Whereas, one year after surgery, the Cobb angle of end plate ring group was (9.4 ± 3.8) degrees, and contral group was (7.5 ± 3.9) degrees, which was significantly lower than that of end plate ring group. Meanwhile, the Dm of end plate ring group was (57.3 ± 2.2) mm, and no end ring group was (55.2 ± 2.6) mm which was significantly lower than that of end plate ring group. The subsidence in end plate ring group was 57.6%, and was 78.9% in no end plate ring group. There was no significant difference in JOA score before and after surgery between two groups. At 1 year after operation, 90.9% (30/33) got excellent or good results in end plate ring group, 89.5% (33/38) got excellent or good results in contral group.

CONCLUSIONThe use of end plate rings could not completely prevent the subsidence of titanium cage, however, which can decrease the occurrence rate of the subsidence and lessen its degree.

Adult ; Aged ; Bone Plates ; Cervical Vertebrae ; pathology ; surgery ; Female ; Humans ; Male ; Middle Aged ; Orthopedic Fixation Devices ; adverse effects ; Spinal Fusion ; instrumentation ; methods ; Spondylosis ; pathology ; surgery ; Titanium

OBJECTIVETo explore the mechanism of miR-124 inhibiting the proliferative activity of prostate cancer PC3 cells.

METHODSLuciferase reporter gene assay was used to examine the specific binding ability of miR-124 to PKM2 mRNA 3'-UTR. After miR-124 was transfected mimic to PC3 cells, the expression levels of PKM2 mRNA and protein were detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, respectively. The effects of miR-124 mimic and PKM2 siRNA on the proliferative activity of the PC3 cells were determined by MTT assay.

RESULTSThe expressions of PKM2 mRNA and protein were upregulated (5.12 +/- 0.35) times and (4.05 +/- 0.20) times respectively in the PC3 cells as compared with those in the RWPE-1 cells (P < 0.05). Luciferase reporter gene assay demonstrated that miR-124 targeted PKM2 3'-UTR. At 24 hours after transfection with miR-124 mimic, the PKM2 protein expression in the PC3 cells was downregulated (0.16 +/- 0.04) times (P < 0.05), while the PKM2 mRNA level was not changed significantly (P > 0.05), as compared with the control group. MTT assay showed that both miRNA-124 mimic and PKM2 siRNA could inhibit the proliferation of the PC3 cells, but the former exhibited a greater inhibitory effect than the latter. After transfection with miR-124 mimic and PKM2 siRNA, the cell growth rates were (66.20 +/- 5.10)% vs (82.10 +/- 6.35)% at 24 hours (P < 0.05) and (49.34 +/- 2.37)% vs (70.10 +/- 5.80)% at 48 hours (P < 0.05).

CONCLUSIONmiR-124 can suppress the proliferation of PC3 cells by regulating the PKM2 gene.

Carrier Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; genetics ; Humans ; Male ; Membrane Proteins ; genetics ; metabolism ; MicroRNAs ; genetics ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; Thyroid Hormones ; genetics ; metabolism ; Transfection

Adult ; Female ; Follow-Up Studies ; Hepatitis B Surface Antigens ; blood ; Humans ; Liver Transplantation ; Male ; Middle Aged ; Tissue Donors

Objective:To investigate the feasibility of detecting excision repair cross-complementing 1(ERCC1)and ERCC2 in peripheral venous blood instead of cancer tissues from esophageal squamous cell carcinoma patients. Methods:The expressions of ERCC1 and ERCC2 mRNA were detected by using RT-PCR in 39 cases of peripheral venous blood samples, esophageal squamous cell carcinoma tissues, and adjacent normal tissues. ELISA was used to determine the levels of ERCC1 and ERCC2 proteins in serum. The periphe-ral blood from 10 healthy volunteers was used as control. Results:Expression levels of ERCC1 and ERCC2 mRNA and protein were significantly higher in peripheral blood from healthy control than those in esophageal carcinoma patients (P<0.05). There was a positive correlation between the expression of ERCC1 and ERCC2 mRNA in peripheral blood and cancer tissues (P<0.01). Conclusion:The expression levels of ERCC1 and ERCC2 mRNA in peripheral blood can indirectly reflect their expression levels in human esophageal squamous cell carcinoma tissues.

OBJECTIVETo evaluate the modified Stoppa approach in treatment of pelvic and acetabular fractures.

METHODSFrom March 2010 to May 2012,16 patients with pelvic fractures and 7 patients with acetabutar fractures were treated by open reduction and internal fixation through the modified Stoppa approach,involving 18 males and 5 females with an average age of 39 years ranging from 17 to 65. By Tile classification, 16 cases of pelvic fractures included 1 case of B1, 2 of B2, 3 of B3, 4 of C1-1, 2 of C1-2, 2 of C1-3, and 1 of C2. By Letournel classification, 7 cases of acetabular fractures included 1 case of anterior column fractures, 1 of transverse fractures, 2 of type T, 1 of anterior column plus posterior transverse fractures, and 2 cases of both columns fractures. For 16 pelvic fractures, the modified Stoppa approach was used exclusively in 9 cases,in combination with the iliac fossa approach in 6 cases, and in combination with the posterior approach in 1 case. For 7 acetabular fractures, the modified Stoppa approach was used exclusively in 4 cases, in combination with the Kocher-Langenbeck approach in 2 cases, and in combination with the Kocher-Langenbeck and iliac fossa approaches in 1 case of both columns fractures.

RESULTSThe average operation time was 130 min (50 to 350 min) and the blood loss averaged 320 ml (100 to 1200 ml). There were no operative complications. The reductions of the pelvic and acetabular fractures were all excellent and good. Twenty-one patients were followed-up from 4 to 24 months (averaged 8 months). The fractures were all healed,the fracture healing time was 2.5 to 5 months (means 3.2 months). Among them, 1 case occurrenced screw loosening, 1 case had mild limited of hip flexion, no case had plate breakage and lateral ventral syndrome.

CONCLUSIONThe modified Stoppa approach can be used to treat pelvic and acetabular fractures effectively, and it has advantages of easy manipulation and a low complication rate.

Acetabulum ; injuries ; surgery ; Adolescent ; Adult ; Aged ; Female ; Fracture Fixation, Internal ; adverse effects ; methods ; Fracture Healing ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Pelvic Bones ; injuries ; surgery