Objective To explore the effects and mechanisms of hepatitis B virus-X protein (HBx) on invasion and migration of hepatocellular carcinoma (HCC) cells.Methods The retrospective cohort study was conducted.The clinicopathological data of 30 patients with liver tumor (20 with HCC and 10 with benign tumor of liver) who were admitted to the Affiliated Hospital of Xuzhou Medical College between July 2014 and July 2015 were collected.HCC tissues of 20 patients with HCC (with history of HBV infection) were collected by surgical resection and peritumoral normal tissues (outside of tumor capsule) of 10 patients with benign tumor of liver (without history of HBV infection) were collected.The expressions of epidermal growth factor receptor 3 (ErbB3)in HCC tissues and peritumoral normal tissues were detected by immunohistochemistry (IHC).The relative expressions of ErbB3 and HBx in HCC tissues and peritumoral normal tissues were detected by Western blot,and relative expressions of ErbB3 in HepG2 of which green fluorescent protein (GFP) and GFP-HBx were respectively transfected were detected.The relative expressions of ErbB3 mRNA in HepG2 transfected by GFP and GFP-HBx were detected by real-time polymerase chain reaction (RT-PCR).The migration and invasion of HepG2 were respectively detected by Transwell assay with and without matrix.The measurement data with normal distribution were represented as $± s.The comparisons between groups were evaluated with the independent-sample t test.Correlation analysis was done by the Pearson test.Results (1) The expressions of ErbB3 were detected by IHC:relative value of mean optical density (MOD) of ErbB3 in HCC tissues of 20 patients with HCC and peritumoral normal tissues of 10 patients with benign tumor of liver were 2.54± 1.33 and O.99±0.29,respectively,with a statistically significant difference (t =6.542,P ＜ 0.05).(2) The relative expressions of ErbB3 and HBx were detected by Western blot:relative expressions of ErbB3 and HBx were respectively 0.79±0.13,1.10±0.28 in HCC tissues of 10 patients with HCC and 1.07±0.17,0 in peritumoral normal tissues of 10 patients with benign tumor of liver,with statistically significant differences (t =3.229,19.486,P＜0.05).The results of Pearson test showed that there was a positive correlation of expression between ErbB3 and HBx in HCC tissues (r=O.637,P＜ 0.05).(3) The relative expressions and transcriptional levels of ErbB3 were detected by Western blot and RT-PCR:relative expressions of ErbB3 in HepG2 of which GFP and GFP-HBx were respectively transfected were O.75±0.11 and 1.10±0.10,respectively,with a statistically significant difference (t=4.291,P＜0.05).The relative expressions of ErbB3 mRNA in HepG2 of which GFP and GFP-HBx were respectively transfected were O.38±0.03 and O.94±0.07,respectively,with a statistically significant difference (t=11.703,P＜O.05).(4) The effects of ErbB3 on migration and invasion of HepG2:numbers of transmenbrane cell in HepG2 of which His and His-ErbB3 were respectively transfected by Transwell assay with matrix were respectively 271± 18 and 463± 31,respectively,with a statistically significant difference (t =8.202,P＜0.05).Numbers of transmenbrane cell in HepG2 of which His and His-ErbB3 were respectively transfected by Transwell assay without matrix were respectively 315±38 and 549±34,respectively,with a statistically significant difference (t =8.310,P＜0.05).Conclusion HBx protein can promote the invasion and migration of hepatocellular carcinoma cells through up-regulating expressions of ErbB3 protein.

In this paper, we summarize the major problems existing in establishing gastric cancer patient-derived xenograft ( PDX) models and its solutions, and introduce its advantages on screening targeted drugs. As many previous re?search emphasized on individual characteristics too much, we argue that we should pay more attention to the generality of what we are studying in order to analyze the genotype of PDX models before taking a targeted therapy.

OBJECTIVETo investigate the clinical application, feasibility and value of 3 T whole-heart contrast enhanced free-breathing navigator-gated three-dimensional coronary magnetic resonance angiography (CE-CMRA).

METHODS3 T CE-CMRA was used to examine patients with suspected coronary heart disease (CAD). Gd-BOPTA (0.2 mmol/kg) was injected intravenously with slow infusion rate (0.3 ml/s) to perform enhancement. Data were post-processed to obtain principal branches of coronary artery and picture quality was evaluated. According to results of selective coronary arteriography (SCAG), the diagnostic accuracy of CE-CMRA for diagnosing CAD was judged by means of detecting significant stenosis (> 50%) of the principal branches based on the 9 segments of coronary artery.

RESULTSTwenty-three out of 26 patients successfully completed the examination. The mean scanning time was (10.4 ± 2.1) minutes, 178 out of 202 (88.1%) SCAG demonstrated segments could be evaluated by CE-CMRA. The imaging quality was superior in proximal and middle segments of coronary artery principal branches than in distal segments. Based on patient-level, there were 9 positive cases and 14 negative cases examined by CE-CMRA compared with 11 positive cases and 12 negative cases examined by SCAG, respectively. The whole diagnose accordance rate of CE-CMRA was 91.3% (21/23) compared with SCAG. The sensitivity, specificity and negative predictive values were 81.8% (9/11), 88.5% (169/191) and 98.8% (9/31) respectively.

CONCLUSIONS3 T CE-CMRA is a feasible non-invasive imaging modality for diagnosing CAD, especially to detect significant stenosis in proximal and middle segments of coronary artery principal branches. However, the detecting efficacy is limited in assessing stenosis of distal segment and small branches of coronary artery.

Aged ; Coronary Angiography ; methods ; Coronary Vessels ; pathology ; Female ; Heart ; diagnostic imaging ; Humans ; Imaging, Three-Dimensional ; Magnetic Resonance Angiography ; methods ; Male ; Middle Aged

OBJECTIVETo evaluate the result of free transmidline bi-lobed scapular skin flap transplantation to repair massive soft tissue defects in upper and lower extremities.

METHODSFree transmidline bi-lobed scapular skin flap was designed according to the characteristics of scapular skin and its vasculature, and they were used to repair massive soft tissue defects in upper and lower extremities as a result of hot crush injury and avulsion injury in 9 patients. The survival of the flap, the blood supply, the color and elasticity of the flap, as well as scar contraction and impairment in function were observed after operation.

RESULTSThe largest flap formed with the horizontal branch and descending branch circumflex scapular artery were 31 x 14 cm and 22 x 16 cm, respectively. The horizontal branch went across the spinal midline for 10 cm. All skin flaps survived with good elasticity and without necrosis. The grafted skin was excellent in the appearance and elasticity, with no scar contracture , and function of the injured extremities recovered well 2 to14 months after the operation.

CONCLUSIONFree transmidline bi-lobed scapular skin flap is an ideal procedure for the repair of massive soft tissue defects in upper and lower extremities.

Adolescent ; Adult ; Female ; Foot Injuries ; surgery ; Hand Injuries ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Surgical Flaps

OBJECTIVETo evaluate the effectiveness of paraumbilical skin flaps for the repair of the severe burn wounds.

METHODSPedicled, island, or free paraumbilical skin flaps, as well as combined transplantation of several kinds of flaps were used to repair the wounds of 39 patients with severe burns. The largest paraumbilical skin flap was 55 x 27 cm(2) in area, with the end approaching the armpit midline of the axilla on the same side, and extending 10cm from the abdominal midline. The survival, color, elasticity of the flaps, degree of deformity resulted from scar contracture, and the condition of donor area were observed.

RESULTSAll the paraumbilical skin flaps survived, without obvious infection subsequent to complete debridement of the wounds. The flaps exhibited good color and elasticity, without scar contracture after 1 to 46 months in 33 patients. No obvious scar formation was observed in the donor area.

CONCLUSIONParaumbilical skin flap is an ideal one for the early repair of deep burn wounds, because it can cover a large area, and can be transferred in different ways according to the area and the position of the wounds.

Adolescent ; Adult ; Burns ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; Skin Transplantation ; methods ; Surgical Flaps ; Thorax ; Umbilicus ; Wound Healing

OBJECTIVETo assess if the modulating effect of platelet-derived growth factor (PDGF)-BB on p21(WAF1) was mediated by upregulating transforming growth factor (TGF)-beta(1) expression in vascular smooth muscle cells (VSMC).

METHODSTGF-beta(1) mRNA and protein expressions were measured by reverse transcription-PCR and ELISA, the protein expressions of p21(WAF1) and the downstream TGF-beta signalling including TGF-beta type I receptor (ALK-5 in VSMC), Smurf2, pSmad2/3, Smad4, Smad7 were detected by Western blot.

RESULTSPDGF-BB significantly upregulated the expressions of TGF-beta(1) at mRNA (0.79-fold) and protein (1.98-fold) levels in VSMC, significantly inhibited the expression of p21(WAF1) (-67 +/- 12)%, and enhanced the expressions of ALK-5, pSmad2/3, Smad4, Smurf2 protein by 1.21-fold, 0.95-fold, 0.69-fold and 2.55-fold respectively, inhibited Smad7 expression (-65 +/- 9)%, these alterations were partially restored by anti-TGF-beta(1) neutralizing antibody.

CONCLUSIONSThese findings suggested that PDGF-BB inhibited p21(WAF1) expression in VSMC partially through upregulating TGF-beta(1) expression via PDGF-BB and TGF-beta signalling pathways.

Animals ; Cell Division ; Cells ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Male ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Platelet-Derived Growth Factor ; pharmacology ; Proto-Oncogene Proteins c-sis ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Transforming Growth Factor beta1 ; metabolism

To investigate the adjuvant effect of plasmid DNA encoding superantigen SEA (D227A) (pmSEA) on immune responses induced by HBV DNA vaccine containing HBV preS2 and S antigen in BABL/c (H-2d). BALB/c mice were immunized intramuscular injection with HBV DNA vaccine (pHBVS2S) mixed with or without pmSEA plasmid. Antibodies againat HBV PreS2 and S antigen in the sera were accessed by Anti-HBs ELISA, and the HBsAg specific cytotoxic T lymphocytes (CTLs) activity was determined by 5 Chromium Release Assay. The HBs peptide-specific IFN-gamma secreting T cells were detected by ELISPOT. Anti-HBs antibody titers and CTLs activity in mice immunized with pmSEA + pHBVS2S group were significant higher (P < 0.05) than pHBVS2S DNA vaccine group. The ratio of IgG1/IgG2a (0.282) was apparently different from the group immunized with peptide (10). Mice immunized with HBV DNA vaccine plus adjuvant produce higher titer of IgG1 and IgG2a antibodies against HBV S antigen 1.36 and 1.73 time higher than that without adjuvant respectively. HBs peptide--specific IFN-gamma secreting T cells increased 2 - 3 times by the pmSEA adjuvant, compared to DNA vaccine group. HBV DNA vaccine (pHBVS2S) induces humoral and cellular immuno-responses in BALB/c mice, and the responses could be significantly boasted by the plasmid encoding mSEA. Therefore the pmSEA was a potential adjuvant for DNA vaccines.
Adjuvants, Immunologic ; Animals ; Enterotoxins ; immunology ; Hepatitis B ; immunology ; prevention & control ; therapy ; Hepatitis B Antibodies ; blood ; Hepatitis B Vaccines ; immunology ; Interferon-gamma ; secretion ; Mice ; Mice, Inbred BALB C ; Staphylococcus aureus ; immunology ; Superantigens ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccination ; Vaccines, DNA ; immunology

OBJECTIVETo explore the therapeutic strategy of hot press injury complicating with blood vessel injury in the upper extremity.

METHODSDecompression procedure was carried out in 8 patients with hot press injury complicating with blood vessel injury in the upper extremities, but the effect was not of satisfactory because there were injuries to brachial, radial and ulnar arteries and also injury to the superficial or deep palmar arch. The blood vessels were repaired with direct anastomosis, anastomosis with flexion of the limb, or transplantation with great saphenous vein, and the wounds were covered with intermediate split-thickness skin graft or pedicled thoraco-abdominal skin flap.

RESULTSOne patient with repair of the brachial artery and intermediate split thickness skin graft received amputation 4 weeks after operation because of lack of soft tissue coverage. The blood supply recovered completely in the other patients, so the affected limbs were saved.

CONCLUSIONPrompt exploration and repair of blood vessel, and coverage of the wound with healthy soft tissue are key procedures for the management of hot press injury complicating with blood vessel injury in the upper extremities.

Adolescent ; Adult ; Arm Injuries ; surgery ; Brachial Artery ; injuries ; Burns ; pathology ; surgery ; Crush Syndrome ; surgery ; Decompression, Surgical ; Female ; Humans ; Male ; Saphenous Vein ; transplantation ; Skin Transplantation ; Wound Healing ; Young Adult

Objective To explore the clinical application value of electrohydraulic shock wave lithotripsy combined with laparoscope and choledochoscope in treatment of intra-and extrahepatic cholangiolithiasis.Methods The clinical data of 42 patients with intra-and extrahepatic cholangiolithiasis treated by laparoscope and choledochoscope(control group) in hepatobiliary surgery department of affiliated hospital of Xuzhou medical university from May 2012 to December 2015,and patients with intra-and extrahepatic cholangiolithiasis treated by electrohydraulic shock wave lithotripsy combined with laparoscope and choledochoscope (combined group) from January 2016 to February 2017,were retrospectively analyzed.The rate of transferring to laparotomy (transfer to laparotomy due to inflammatory adhesion or difficulty to remove),residual rate of choledocholithiasis,incidence of bile leakage and incidence of common bile duct stricture between two groups were compared.Results All patients in both groups were recovered.In the control group,there were 6 cases(6/42) transferred to laparotomy,among which 2 cases were transferred to laparotomy due to severe abdominal inflammatory adhesion and 4 cases were due to the difficulty to remove the calculus,without bile leakage or common bile duct stricture.There were 3 cases with residual choledocholithiasis,but without bile leakage or common bile duct stricture.In the combined group,there were 2 cases(2/42) transferred to laparotomy due to severe abdominal inflammatory adhesion and 5 cases used the technique of electrohydraulic shock wave lithotripsy without post-operation residual choledocholithiasis,bile leakage or bile duct stricture.In the control group,the residual choledocholithiasis was removed with T tube fistula choledochoscope.Conclusion In the process of minimally invasive surgery to treat intra-and extrahepatic cholangiolithiasis,electrohydraulic shock wave lithotripsy under laparoscope and choledochoscope can decrease the incidence of transferring to laparotomy due to the difficulty to remove calculus and the residual rate of calculus.It can be applied safely and efficiently to patients with intra-and extrahepatic cholangiolithiasis but with laparoscopic surgery indications.

BACKGROUND:Icariin can promote bone formation and inhibit bone resorption,but it is difficult to dissolve in water,and its bioavailability is very low in vivo.Therefore,a suitable carrier is essential to fully utilize the biological activity of icariin.OBJECTIVE:To prepare an icariin/β-tricalcium phosphate (β-TCP) composite scaffold and to characterize its biological characteristics.METHODS:Three-dimensional printing technology was utilized to prepare porous β-TCP scaffolds carrying nano zinc oxide,and the compressive strength of porous β-TCP scaffolds was detected before and after addition of nano zinc oxide.Water absorption and porosity of the porous β-TCP scaffolds were also measured.Ultrasonic emulsification solvent dialysis was performed to prepare icariin/poly(lactic-co-glycolic acid) (PLGA) microspheres,followed by the detection of water absorption and porosity.The porous β-TCP scaffolds and icariin/PLGA microsphere suspension were mixed to prepare icariin/β-TCP composite scaffolds.Microstructure observation of the composite scaffolds was done by scanning electron microscope,and meanwhile,water absorption and porosity were detected.The composite scaffold was then immersed into PBS,and icariin concentration in the supematant was measured at corresponding time points,based on which Icariin cumulative release curve was drawn.RESULTS AND CONCLUSION:(1) The porous microstructure of the porous β-TCP scaffold was regular,well distributed,and the connectivity was good.The pore spacing was about 600 μm.After addition of nano zinc oxide particles,the surface structure of the scaffold was more compact and the crystallinity was higher.(2) PLGA microspheres were spherical with a diameter of 1-4 μm,and the microspheres were uniform in size.(3) The maximum compressive strength of the porous beta tricalcium phosphate scaffold was (2.98±0.78) MPa,and increased to (8.95±0.29) MPa after addition of nano zinc oxide.(4) The water absorption rate and porosity were (25.09±0.96)％ and (66.93±2.84)％ for the porous β-TCP scaffold,(28.46±1.85)％ and (32.65±3.32)％ for the icariin/β-TCP composite scaffold,respectively.(5)The average encapsulation efficiency of the PLGA microspheres was (78.87±2.31)％,and the drug loading was (6.04±1)％.(6) The release amount of icariin could reach 52％ of the total amount at 16 days and 60％ of the total amount at 32 days.These findings indicate that the icariin/β-TCP composite scaffold has good mechanical properties and sustained-release performance.