BACKGROUND: Bone morphogenetic proteins (BMPs) are involved in the formation of various tissues including bone, cartilage, tendon, and ligament. Vascular endothelial growth factor (VEGF) promotes angiogenesis by increasing the permeability and migration of endothelial cells.OBJECTIVE: To construct a co-expressing vector of human bone morphogenetic protein 2 (BMP2) and vascular endothelial growth factor 165 (VEGF165), and observe the expression of BMP2 and VEGF165 in human bone marrow mesenchymal stem cells (hBMSCs).DESIGN: Observation control trail.SETTING: Department of Plastic Surgery, Affiliated Hospital of Luzhou Medical College and Plastic Surgery Hospital of Peking Union Medical College, Chinese Academy of Medical Sciences.MATERIALS: The MSCs derived from the healthy adult volunteers of marrow donors. pIRES-EGFP-hVEGF165 containing total length of cDNA sequence of human VEGF165 gene was provided by Dr. Cheng Ting from Plastic Surgery Hospital of Peking Union Medical College. pSP65-hBMP2 containing total length of cDNA sequence of human BMP2 gene was provided by Dr. Guo Ximin from the Academy of Military Medical Sciences. Enkaryotic expression vector pIRESneo (Clontech Company) Pyrobest DNA Polymerae, restriction enzyme, DNA ligase and plasmid extraction kit, DNA Fragment Purification Kit (TaKaRa Company), LiorfectamineTM liposome transfection kit, DMEM medium, trypase, TRIzoIRNA extraction kit (Gibco BRL), Omniscript RT kit (Qiagen), TaqplusDNA polymerase (Promega), PMSF, leupeptin, aprotinin, chymostatin (Sigma), protease inhibitor, PVDF membrane (Amersham-Pharmacia Biotech), rabbit anti-human BMP2 antibody and VEGF monoclonal antibody (Santa Cruz Company), goat anti-rabbit lgG-peroxydase (Wuhan Boster), G418 (Ameresco Company in U.S).METHODS: The experiment was conducted in the Affiliated Hospital of Luzhou Medical College and the Plastic Surgery Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences between June 2005 and April 2006.hBMP2 and hVEGF165 cDNA were directional cloned into multiple clone sites of the eukaryotic expression vector pIRESneo. The recombinant plasmid was identified by restrictive enzyme Xho Ⅰ/Bgl Ⅱ digestion analysis and DNA sequencing. Liposome-mediated gene transfer method was used to transfect hBMSCs. For observation, the transfected cells were divided into IRES-hBMP2-VEGF165 group, pIRES-hBMP2 group, pIRES-VEGF165 group and empty vector group, which were transfected with pIRES-hBMP2-VEGF165, pIRES-hBMP2, pIRES-VEGF165 and pIRES-neo. Meanwhile, the same number nontransfected cells were selected as blank control group. The reverse transcription polymerase chain reaction (RT-PCR) and Weszern blot were employed to observe the expression and secretion of hBMP2 and hVEGF165 gene and protein.expression vector hBMP2 and hVEGF165 gene sequence were the same as reported after restrictive enzyme EcoRI and Bgl Ⅱ digestion analysis and pIRES-BMP2 gene sequencing, which showed that the recombinant plasmid pIRES-hBMP2-VEGF165 highly expressed hBMP2-mRNA and VEGF165-mRNA, but the non-transfected or transfected with pIRES-hBMP2-VEGF165 or pIRES-hBMP2 secreted a great quantity of hBMP2, but that non-transfected or transfected with pIRES-VEGF165 or empty vector secreted only little.CONCLUSION: The co-expressing vector of hBMP2 and hVEGF165 can be expressed stably in hBMSCs.

Phosphoinositide 3-kinase/serine-threonine kinase (PI3K/AKT)has been found playing an important role in the pathogenesis of severe acute pancreatitis (SAP)in recent years,but the underlying mechanism has not been clarified.Aims:To investigate the role of PI3K/AKT in regulating the inflammatory response in SAP by evaluating the effect of insulin-like growth factor-Ⅰ (IGF-Ⅰ)and wortmannin,the agonist and inhibitor of PI3K/AKT on Toll-like receptor 4 (TLR4)signaling pathway in macrophage cell line RAW264.7.Methods:RAW264.7 cells were treated with different concentrations of lipopolysaccharide (LPS ), IGF-Ⅰ and wortmannin, respectively, and cell viability was determined by CCK-8 assay.RAW264.7 cells were divided into blank control group (no treatment),LPS group (LPS 1 μg/mL),IGF-Ⅰ group (IGF-Ⅰ 100 ng/mL +LPS 1 μg/mL),wortmannin group (wortmannin 100 nmol/L +LPS 1 μg/mL)and IGF-Ⅰ +wortmannin group (wortmannin 100 nmol/L +IGF-Ⅰ 100 ng/mL +LPS 1 μg/mL).Protein expressions of tumor necrosis factor-α(TNF-α)and interleukin-6 (IL-6)were detected by ELISA;mRNA expressions of TLR4,myeloid differentiation factor 88 (MyD88),AKT,PI3K,p38 mitogen-activated protein kinase (p38MAPK)and nuclear factor-κB (NF-κB)were determined by real-time PCR.Results:After treated with LPS,IGF-Ⅰand wortmannin, respectively,no differences in cell viability of RAW264.7 cells were found between different concentrations groups (P>0.05).Protein expressions of TNF-αand IL-6 in LPS,IGF-Ⅰ,wortmannin and IGF-Ⅰ +wortmannin groups were significantly higher than those in blank control group (P<0.05 ).Protein expressions of TNF-αand IL-6 in wortmannin group were significantly lower than those in LPS and IGF-Ⅰ groups (P<0.05),and those in IGF-Ⅰ+wortmannin group were significantly lower than those in IGF-Ⅰ group (P<0.05).In LPS group,mRNA expressions of AKT and PI3K as well as TLR4 and its downstream molecules MyD88,p38MAPK and NF-κB were significantly higher than those in blank control group (P <0.05 ).Expressions of all above-mentioned mRNAs in IGF-Ⅰ group were further increased and significantly higher than those in LPS group (P<0.05).Expressions of all above-mentioned mRNAs in wortmannin group were significantly lower than those in LPS and IGF-Ⅰ groups (P<0.05 ),and those in IGF-Ⅰ+wortmannin group were significantly higher than those in wortmannin group (P<0.05),but significantly lower than those in IGF-Ⅰ group (P<0.05).Conclusions:PI3K/AKT might regulate TLR4 signaling pathway and its downstream molecules in macrophages, thereby affects the expressions of inflammatory cytokines and being involved in the pathogenesis of inflammatory response in SAP.

BACKGROUND: Enophthalmos deformity is the most common complication caused by orbital blow-out fractures, and others are diplopia and worsening of visual acuity. Since the therapeutic result of orbital blow-out is not satisfactory and many complications exist after operation, it is still a dispute to select implantation materials and therapeutic regimens.OBJECTIVE: To observe the therapeutic effect and assess the improvement of visual function by surgical reconstruction with porous high-density polyethylene (Medpor) for the correction of enophthalmos deformity caused by orbital blow-out fractures.DESIGN: A pre-and postoperative controlled study.SETTING: Beauty Center for Trauma Repair,Plastic Surgery Hospital,Peking Union Medical College, Chinese Academy of Medical Science .PARTICIPANTS: Totally 56 patients with orbital blow-out fractures who had enophthalmos deformity caused by fists or traffic accidents, treated at Beauty Center for Trauma Repair,Plastic Surgery Hospital, Peking Union Medical College, Chinese Academy of Medical Science, were selected in this study from December 1996 to March 2004. Final diagnosis were made with case history, X-ray film, two-demensional and three-dimensional CT before operation. 24 cases were accompanied with other areas fractures such as zygoma and nasal bone, 34 cases with diplopia, 35 cases with visual acuity worsening after injured.METHODS: ①Material implantation: Exposure of the orbital floor, inferior and medial walls could be performed through a 2 mm inferior subciliary incision of 3 cm long. To approach the orbital rim via a dissection plane anterior to the orbital septum, sub-periosteal dissection was then performed over the orbital rim, and along the orbital floor to the orbital apex. Mobilized the soft tissue from the bone throughout the entire area of fractures and re-position it to its proper position. Took Medpor (Type 6331) sheets as the implantation materials, trimmed Medpor sheets according to the radian and anatomic form, and 2 mm larger than the defect rim was needed.If other operations were needed during the operation, they could be done.Mannitol and dexamethasone should be used just postoperatively to decrease edema of the orbital contents and reduce inner orbital excessive pressure. ②Functional evaluation standard: Diplopia: completely disappear meant recovered, less diplopia residual meant improvement, no improvement meant inefficacy. Enophthalmos: marked improvement meant the degree of enophthalmos stabilizated at below 2 mm, less improvement meant stabilizated at above 2 mm.MAIN OUTCOME MEASURES: ①Improvement of enophthalmos; ②Improvement of diplopia ; ③Improvement of visual acuity.RESULTS: ①All 56 cases of enophthalmos deformities caused by orbital blow-out fractures improved greatly. ② Of all the 34 patients with diplopia, 27 recovered. ③ 9 patients' visual acuity of 35 improved with different degrees. No diplopia or visual acuity worsening occurred. With a follow-up ranging from 2 months to 5 years, the degree of enophthalmos stabilizated at below 2 mm, and no relapse and other complications occurred.CONCLUTION: Medpor has such advantages as better histocompatibility,fewer complications and better visual function improvement, so it is the preferred implantation material for correcting enophthalmos deformity caused by orbital blow-out fractures.

BACKGROUND: Fibular fixation and tantalum rod implantation are two commonly used methods for the treatment of early osteonecrosis of the femoral head (ONFH), both of which can effectively delay or even reverse the progress of ONFH. However, further comparative evaluation on their mechanical properties and therapeutic efficacy is required.OBJECTIVE: To compare the clinical efficacy of fibular fixation and tantalum rod implantation on ONFH at early stage.METHODS: Fifty-eight patients (81 hips) suffered from ONFH with ARCO stage 1 and stage 2, and underwent fibular fixation (30 cases, 41 hips) or tantalum rod implantation (28 cases, 40 hips). Postoperatively, both groups were followed up for over 2 years. The Harris scores of the hip were compared between two groups before and after treatment. With femoral head collapse and the collapse distance > 4 mm as observation points, the survival rate of the femoral head was compared between two groups.RESULTS AND CONCLUSION: The postoperative Harris scores of the two groups were significantly improved than before (P < 0.05). With the appearance of femoral head collapse as the observation point, the Kaplan-Meier survival curve showed that the overall survival rate of the hip was 83% in the fibular fixation group and 65% in the tantalum rod implantation group. After examined by log-rank (Mantel-Cox), there was a significant difference in the survival rate of the hip at Stage IIC between two groups (P=0.0431). With > 4 mm collapse as the observation point, the Kaplan-Meier survival curve showed that overall survival rate of the hip was 95% in the fibular fixation group and 83% in the tantalum rod implantation group. After examined by log-rank (Mantel-Cox), there was a significant difference in the survival rate of the hip at Stage IIC between two groups (P=0.0418). To conclude, both fibular fixation and tantalum rods implantation applied to ONFH at early stage can effectively improve the hip function, and the survival rate of the hip at ARCO Stage IIC is better in patients undergoing fibular fixation than tantalum rod implantation.

OBJECTIVETo study the combination of Mandibular distraction and orthognathic techniques for the reconstruction of adult hemifacial microsomia.

METHODSThe three-dimensional CT reconstruction data was used with Mimics for preoperation design. The osteotomy location, distraction vector, distraction distance were decided before operation with a surgical guider. At the first stage, internal distractor was implanted after ostetomy through an extra-oral approach. The distraction begun 5-7 days after operation with a frequency of 1 mm/day. After distraction, the distractor was maintained for 3-6 months. At the second stage, the distractor was removed. Le Fort I osteotomy was performed in order to correct the cross-bite and improve the facial contour. Usually, bone graft was inserted into the gap after Le Fort I osteotomy. The genioplasty was also performed if necessary.

RESULTS9 cases of adult hemifacial microsomia with severe mandibular deviation were treated. The facial asymmetry were improved greatly. 1 patient suffered an wound infection in the maxillary region after Le Fort I osteotomy and healed uneventfully with wound irrigation.

CONCLUSIONSMandibular distraction combined with orthognathic surgery is an effective procedure for adult hemifacial microsomia with complicated mandibular hypoplasia.

Adult ; Aged ; Bone Transplantation ; Facial Asymmetry ; surgery ; Goldenhar Syndrome ; surgery ; Humans ; Mandible ; surgery ; Osteogenesis, Distraction ; methods ; Osteotomy, Le Fort ; methods

Objective to investigate the efficacy of moxibustion treatment to rheumatoid arthritis(RA) patients and the impact to blood levels of rheumatoid factor(RF) ,C reactive protein(CRP) ,and interleukin-6(IL-6) .Methods RA patients were divided in-to treatment group and comfort group .The two groups were treated by moxibustion and sham-moxibustion for 2 courses ,respec-tievely .Results Overall clinical efficacy rate was higher in treatment group than in comfort group(P<0 .01) .The arthralgia ,ar-throcele ,tenderness and its degrees ,and morning stiffness time were significant improved in both groups(P<0 .05) ,and the indexes were better in treatment group than comfort group(P<0 .05) .The blood levels of RF ,IL-6 and CRP were reduced after treatment in both groups(P<0 .05) ,and which of treatment group reduced more than those of comfort group(P<0 .05) .Conclusion Moxi-bustion treatment can significantly improve clinical outcomes of RA and markedly reduce the blood levels of RF ,IL-6 and CRP of RA patients .

Humans ; Male ; Middle Aged ; Positron-Emission Tomography ; Situs Inversus ; complications ; pathology ; Thymoma ; diagnosis ; pathology

A method for quantitative detection of florfenicol by colloidal gold lateral flow immunoassay was developed.The experimental conditions including pH value, concentrations of antibody in the process of conjugation between the colloidal gold and antibody, amount of gold-labeled antibody, concentration of the antigen sprayed on test lines (T line), and detection time were optimized.With a colloidal gold strip reader, the signal intensity of T lines and control lines (C line) on lateral flow strips was recorded.The T/C ratio of negative control and positive samples was defined as B0 and Bx, and the standard curve was established by plotting the Bx/B0 ratio against the concentration of florfenicol.This assay showed a good linear range from 0.1 to 1.5 ng/mL with the limit of detection of 0.08 ng/mL, while the result could be obtained within 15 min.The result showed that this quantitative method was convenient and rapid, and could be used in screening a large amount of samples on site.

BACKGROUNDThe expanded endonasal approach (EEA) is used sparingly by surgeons for resection of lesions in the ventrocranial base. Herein, we examined the anatomy of the ventrocranial base by endoscopy and comment on the use of EEA in clinical practice.

METHODSTwenty artery-injected adult cadaveric heads were studied under surgical conditions using the endoscopic EEA. The extent of the surgical exposure, the endoscopic anatomic view and the maneuverability of surgical instruments about the suprasellar region were studied by the endoscopic EEA.

RESULTSThe EEA by endoscope can reach the suprasellar region. In this approach, the optocarotid recess, supra and infra-optic chiasm interspace, the ophthalmic artery and others were important anatomical landmarks for identification of the suprasellar region.

CONCLUSIONSThe endoscopic EEA can be used to remove many types of lesions in the ventrocranial base. The microanatomy observed using the endoscope provides important anatomical information on the suprasellar region for neurosurgeons.

Endoscopy ; Humans ; Nasal Cavity ; anatomy & histology ; Sphenoid Sinus ; anatomy & histology

OBJECTIVETo study the extent and incidence of sensorineural hearing loss (SNHL) after radiotherapy (RT).

METHODSTwenty-eight patients with diagnosed nasopharyngeal carcinoma (NPC) were selected. The pure-tone audiography, auditory brain stem evoked response (ABR), impedance audiometry and evoked otoacoustic emissions (EOAE) recordings were performed before RT, 1 month, 1, 2 and 5 years after RT.

RESULTSAt 1 month after RT, there were 7.1 and 25.7 dB increased mean bone conduction (BC) thresholds at speech (0.5 - 4.0 kHz) and at high frequency (8.0 kHz), and their BC thresholds were statistically significant increase than those before RT, respectively (P < 0.001). At 1 year after RT, there were 17.6 and 28.1 dB increased respectively, and their thresholds were statistically significant increase than those at pre-irradiation (P < 0.001). There were also significant increases in thresholds than those at 1 month of post-irradiation (P <0.001 or P < 0.05). At 2 years after RT, 21 and 27.4 dB were increased at respective those two frequencies, and there was a statistically difference only at speech frequencies when compared with those at 1 year after RT (P < 0.05). At 5 years after RT, 26.7 and 35.8 dB were increased at these two frequencies, and there were significant increases in threshold than those before, 1 month, 1 and 2 years after RT, respectively (P < 0.001). From 1 month to 5 years after RT, 37. 5% to 94. 7% of ears had a BC hearing threshold of at least 15 dB losses at speech frequency, whereas the percentage at high frequency was 85.4 to 97.4%. Up to 63.2% and 73.7% of ears had 30 dB SNHL at least at speech and high frequency, respectively. Furthermore, the degree of mean threshold loss was greater at high frequency than at speech frequency. The mean value of wave I, III and V latency, and I -V interpeak latency intervals of ABR had no significant difference between at 1 month after RT and before RT (P > 0.05). The wave I , III and V latency, and I - V interpeak latency intervals at 1 year and 2 years were significantly prolonged when compared with those before and 1 month after RT (P < 0.05), but there were no significant difference between 1 year and 2 years after RT (P > 0.05). The wave I, III and V latency, and I -V interpeak latency intervals at 5 years after RT were also significantly longer than those before RT (P < 0.001). There were significant difference in wave I , III and V latency (P < 0.05), and no significant difference in wave I - V interpeak latency intervals (P > 0. 05) between 5 years after RT and 1 year or 2 years after RT. Seven of 10 ears at 1 year after RT and 4 of 7 ears at 5 years after RT had normal EOAE, but they all had abnormal ABR response.

CONCLUSIONSSNHL in NPC patients start soon after completion of RT, especially more commonly in high frequency. The incidence and the extent of hearing loss are increased with time of follow-up. The hearing impairment could occur in the cochlea and/or the retrocochlear auditory pathway, which show that the sensitivity of radiation damage may be different in different patient and anatomic site of auditory system.

Adult ; Audiometry ; Evoked Potentials, Auditory, Brain Stem ; Female ; Hearing Loss, Sensorineural ; etiology ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; radiotherapy ; Otoacoustic Emissions, Spontaneous ; Radiotherapy ; adverse effects