Ear ; surgery ; Humans ; Parotid Neoplasms ; surgery ; Surgical Procedures, Operative ; methods

OBJECTIVETo explore the effect of electroporation mediated gene therapy on bone mineral density and strength of new-formed bone in mandibular distraction gap, so as to enhance the osteogenesis and shorten the distraction term.

METHODSNew-Zealand rabbits were employed. The distraction began after 3 days of latency period at the rate of 0. 8 mm per day for 7 days. After distraction, the rabbits were randomly divided into 5 groups to receive injection in the distraction gap with recombinant plasmid 2 microg (0.1 microg/microl) pIRES-hVEGF165-hBMP2 in group A, with recombinant plasmid pIRES-hBMP2 in group B, with recombinant plasmid pIRES-hVEGF165 in group C, with pIRES in group D, and with normal saline (NS) in group E. After injection, electroporation was performed in all the groups. After 1 week, 2 weeks, 4 weeks and 8 weeks of consolidation, all the animals underwent X-ray and quantitative computed tomography (QCT). The new-formed bone in distraction gap was selected as regions of interest (ROI) to measure the bone mineral density(BMD). Then the rabbits were sacrificed and the new-formed bone samples were harvested to detect 3-point crushing strength.

RESULTSBMD of newly formed bone in group A, B and C was markedly higher than that in group D and E (P < 0.01). After 2 weeks of consolidation, BMD in group A was much higher than that in the other groups, but there was no difference between group B and C. After 4 weeks of consolidation, BMD in group A and B was markedly higher than that in group C, D and E (P < 0.01). After 8 weeks of consolidation, BMD in group A was markedly higher than that in the other groups. While the BMD was not significantly different between group B and C, but the BMD in group B and C was higher than that in group D and E (P < 0.01). After 4 weeks of consolidation, the 3-point crushing strength of newly formed bone in group A was markedly higher than that in group B,C, D and E (P < 0.01), which was still the same after 8 weeks of consolidation. And the crushing strength in group B was higher than that in group C, D and E (P < 0. 05).

CONCLUSIONSElectroporation-mediated transfection of recombinant plasmid pIRES-hVEGF165-hBMP2 could greatly enhance osteogenesis and calcification. A combination of VEGF and BMP may promote osteogenesis and angiogenesis simultaneously, so as to magnify the effect of each growth factor, resulting a synergetic effect.

Animals ; Bone Density ; Bone Regeneration ; Electroporation ; Genetic Therapy ; Mandible ; physiology ; surgery ; Osteogenesis, Distraction ; Rabbits

OBJECTIVETo investigate the effect of gene transfection at different time on bone mineral density and strength of newly formed bone in mandibular distraction gap in rabbit, so as to explore the optimal time for gene therapy and enhance the therapeutic effect.

METHODS48 New-Zealand rabbits were employed to receive mandibular osteotomy and implantation of distraction devices bilaterly. Then the rabbits were randomly divided into 4 groups as group A, B and C and D. The animals in group A, B, and C were transfected with recombinant plasmids pIRES-hBMP2-hVEGF165 via electroporation-mediated approach at latency period, distraction period, consolidation period respectively. Group D was used as control group without gene transfection. After 3 days of latency period, the distraction devices were activated at the rate of 0.8 mm per day for 10 days. Three rabbits in each group were sacrificed at 1 wk, 2 wk, 4 wk and 8 wk of consolidation respectively. The mandibles were harvested and the left mandible received X-ray examination for bone healing, and quantitative computed tomography (QCT) dectection for the bone mineral density (BMD) of newly formed bone in the distraction gap. The biomechanical properties of the new generation bone at 4 th and 8 th week of consolidation in each group were detected by three point bending test.

RESULTSThe bone mineral density and the biomechanical strength of newly formed bone increased along the length of consolidation in each group. After 1 week of consolidation, there was no significant difference in BMD among group A (83.43 +/- 9.96), group B (92.29 +/- 11.25), group C (89.93 +/- 14.15), P > 0.05. But the BMD of group A, B and C was higher than that of group D (70. 31 +/- 3.30), P < 0.05. After 2wk, 4 wk and 8 wk of consolidation, the BMD of group B (137.54 +/- 7.20,492.93 +/- 17.57, 790.48 +/- 12.19) was significantly higher than those of group A (121.44 +/- 9.27, 396.15 +/- 15.70, 603.39 +/- 16.46), C (125.06 +/- 7.24, 464.15 +/- 15.45, 764.15 +/- 17.28), and D (98.86 +/- 8.13, 336.45 +/- 11.95, 577.89 +/- 18.43), P < 0.05. The biomechanical parameters were also higher in group B than those of group A, C and D after four and eight weeks of consolidation (P < 0.05).

CONCLUSIONIt is better to transfect gene at the beginning of distraction (distraction period) than at other stages of DO. In this way, more remarkable effect could be obtained on new bone formation. It suggests that the distraction stage is the optimal time for gene therapy.

Animals ; Bone Density ; genetics ; physiology ; Electroporation ; Genetic Therapy ; Mandible ; physiology ; surgery ; Osteogenesis, Distraction ; Osteotomy ; Rabbits ; Time Factors ; Transfection

[Objective]To study the intervention effect of mild hypotherrnia on cerebral edema in rats after cerebral infarction.[Methods]Twelve male SD rats were randomized into control group (n--6) and mild hypotherma treatment group (n=6).The rat models of focal cerebral infarction were established with obstructing the middle cerebral artery of rats by suture emboli.Rats of the mild hypotherma treatment group were treated by mild hypotherm for 3 h after the models were established.MRI was performed at 6 h,and 1,3,5 and 7 d after the success of model making.Cerebral infarction volume,absorption rate of edema,and signal intensity ratios (SIRs) ofTIWl,T2WI,FLAIR and DWI sequencas in the infarction zones,and relative variance ratio of SIR (△SIR) were measured and calculated.[Results] In the mild hypotherma treatment group,the cerebral infarction volumes 3,5 and 7 d after the models were established were significantly decreased as compared with those in the control group (P＜0.05);the absorption rate of edema (the 7th d against the 3th d of infarction) in the the mild hypotherma treatment group was significantly higher than that in the control group (P＜0.05).Three,5 and 7 d after infarction,the SIRs of TIWI were all obviously higher than those in the control group,and the SIRs of T2WI and FLAIR were all significantly lower than those in the control group (P＜0.05).The SIR of DWI in the mild hypotherma treatment group was obviously lower than that in the control group 7 d after infarction (P＜0.05).The △SIRs of T2Wl,FLAIR and DWI in the mild hypotherma treatment group were significantly higher than those in the control group (P＜0.05).[Conclusion] Mild hypothermia has a significant inhibition effect on cerebral edema in rats after cerebral infarction.

To solve the issues of costly planting of facility cultivation method and inferior efficacy than wild herbs of Dendrobium officinale, the cliff epiphytic cultivation method was studied. To research the growth, agronomic traits, yield, polysaccharide and alcohol-soluble extract contents were measured on the D. officinale from different water regulation and cliff slope gradients treatments. The results showed that D. officinale epiphytic at 85 degrees-90 degrees cliff and sprayed water 1-2 h x d(-1) at the growing season can get better growth and obtain high yield, and the morphology has no different from wild cliff D. officinale, even in the environments without shade. The contents of polysaccharide and alcohol-soluble extract are closely related to the physiological ages, but significantly higher than the facility cultivation. It is possible that environmental stresses benefit the accumulation of polysaccharides, alcohol-soluble extract and other efficient ingredients.
Agriculture ; methods ; Dendrobium ; chemistry ; growth & development ; Drugs, Chinese Herbal ; analysis ; Polysaccharides ; analysis ; Water ; analysis

OBJECTIVETo explore the effect of compound qizhu granule (CQG) on cellular immunity of chronic hepatitis B (CHB) patients.

METHODSTotally 103 CHB patients treated with lamivudin (LAM) for 6 months, who had partial virological response (HBeAg positive) were randomly assigned to two groups, 50 in the treatment group and 53 in the control group. All patients took LAM 100 mg (once a day) plus ADV 10 mg (once a day). Patients in the treatment group additionally took CQG, one dose per day. After one-year treatment hepatitis B virus (HBV) DNA negative rates, HBeAg seroconversion, levels of HBV specific cytotoxic T lymphocyte (CTL), non-specific CTL and natural killing (NK) cells were compared between the two groups.

RESULTSAfter 1-year treatment, HBV DNA negative rate of the treatment group was 88: 0% in 44 cases, slightly higher than that of the control group (41 cases, 77.4%), but with no statistical difference (P >0.05). HBeAg seroconversion of the treatment group was 32.0% in 16 cases, higher than that of the control group (8 cases, 15.1%), with statistical difference (P <0.05). Levels of HBV specific CTL (0.79%±0. 07%), non-specific CTL (19.4%±1.8%) and NK cells (14. 1%± 1.5%) of the treatment group were higher than those of the control group (0.58% ± 0.08%, 17.5% ± 1.7%, and 11.1%±1.5%, respectively; allP <0.01).

CONCLUSIONTreating CHB patients with partial virological response by ADV plus CQG could improve specific and non-specific cellular immunity, thereby elevating HBeAg seroconversion rate.

Drugs, Chinese Herbal ; therapeutic use ; Hepatitis B e Antigens ; immunology ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Immunity, Cellular ; immunology ; T-Lymphocytes, Cytotoxic ; drug effects

OBJECTIVETo evaluate therapeutic effects of Wallis interspinous dynamic stabilization in treating ASD after lumbar spinal fusion.

METHODSTotally 40 patients (included 16 males and 24 females, aged 25 to 60 years old) with degenerative disc disease were treated with posterior interbody fusion. Among them, 20 cases (treatment group) were treated with posterior interbody fusion combined with Wallis interspinous dynamic stabilization, while other 20 cases (control group) only treated with posterior interbody fusion. JOA score and VAS score were compared after inserted Wallis interspinous dynamic stabilization at 1 month and 3 years, and changes of intervertebral disc height of adjacent segment and cross-sectional area of the canal were tested and compared.

RESULTSAll patients were followed up from 3 to 5 years with an average of 3.6 years. All injuries were healed at stage I and the pain were released after treatment. There were no significant meaning in JOA score and VAS score at 1 month after treatment between two groups (P>0.05), while had meaning at 3 years (P<0.05). There were no statistical significane in intervertebral disc height of adjacent segment and cross-sectional area of the canal at 1 month after treatment (P>0.05), while had statistical meaning at 3 years (P<0.05).

CONCLUSIONThere is no difference in immediate effects between two groups. Both of them can obtain good results for effective decompression. Medial-term effectiveness of treatment group is obviously better than control group, which depends on Wallis interspinous dynamic stabilization to plays good biology effects and effective accelerate adjacent degeneration caused by lumbar fusion.

Adult ; Decompression, Surgical ; Female ; Humans ; Intervertebral Disc Degeneration ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Fusion ; Treatment Outcome

OBJECTIVETo establish an artificial somatic-autonomic reflex arc in rats and observe the following distributive changes of neural fibers in the bladder.

METHODSAdult Sprague-Dawley rats were randomly divided into three groups: control group, spinal cord injury (SCI) group, and reinnervation group. DiI retrograde tracing was used to verify establishment of the model and to investigate the transport function of the regenerated efferent axons in the new reflex arc. Choline acetyltransferase (ChAT) in the DiI-labeled neurons was detected by immunohistochemistry. Distribution of neural fibers in the bladder was observed by acetylcholine esterase staining.

RESULTSDiI-labeled neurons distributed mainly in the left ventral horn from L3 to L5, and some of them were also ChAT-positive. The neural fibers in the bladder detrusor reduced remarkably in the SCI group compared with the control (P < 0.05). After establishment of the somatic-autonomic reflex arc in the reinnervation group, the number of ipsilateral fibers in the bladder increased markedly compared with the SCI group (P < 0.05), though still much less than that in the control (P < 0.05).

CONCLUSIONThe efferent branches of the somatic nerves may grow and replace the parasympathetic preganglionic axons through axonal regeneration. Acetylcholine is still the major neurotransmitter of the new reflex arc. The controllability of detrusor may be promoted when it is reinnervated by the pelvic ganglia efferent somatic motor fibers from the postganglionic axons.

Acetylcholinesterase ; biosynthesis ; Anastomosis, Surgical ; Animals ; Autonomic Fibers, Preganglionic ; physiology ; Cholinergic Fibers ; metabolism ; Immunohistochemistry ; Motor Neurons ; cytology ; metabolism ; Nerve Regeneration ; physiology ; Neural Pathways ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reflex ; physiology ; Spinal Cord Injuries ; physiopathology ; Spinal Nerve Roots ; surgery ; Urinary Bladder ; innervation ; physiology ; surgery ; Urinary Bladder, Neurogenic ; surgery

OBJECTIVETo investigate flap survival after transfection using gene encoding vascular endothelial growth factor (VEGF).

METHODSIn 30 Sprague-Dawley rats, the anterior abdominal skin flap supplied by the epigastric vessels was created. The animals were divided into three groups, with ten of each. The first group was treated with a mixture of liposomes and the cDNA encoding the 165-amino acid isoform of VEGF; the second group was treated with control blank plasmid DNA and liposome transfection medium; the third group was treated with physiological saline. Four days after injection, the epigastric artery and vein were ligated and the blood flow in the flap was evaluated by intraperitoneal injection of fluorescence solution. Seven days later, the survival area of the flap was measured by planimetry. After the animals were killed, specimens were harvested from the anterior abdomen skin flap for immunohistological evidence of VEGF expression and for hematoxylin and eosin staining of microvascular growth.

RESULTS30 minutes after pedicle ligation the average fluorescence staining planimetry of the three groups (PCD-VEGF165, PCD and physiological saline) was 60.64%, 30.15% and 29.89% respectively. Tissue survival planimetry of the three groups was 92.3%, 30.5%, 31.8%. There was significant difference between the first group and the latter two (P < 0.05). Immunohistochemical staining documented increased deposition of VEGF cDNA in the first group compared to the control groups (P < 0.05). Normal staining documented that the average vessel number of the three groups was 101.72, 91.35 and 89.85 (P < 0.05), the average vessel lumen diameter was 26 microns, 31.09 microns and 32.51 microns(P < 0.05).

CONCLUSIONThrough administration, PCD-VEGF165 can transfect the anterior abdominal skin flap and enhance its survival. There was express of VEGF protein in the treated flap.

Animals ; Biomarkers ; metabolism ; DNA, Complementary ; administration & dosage ; Epigastric Arteries ; Genetic Therapy ; Graft Survival ; physiology ; Liposomes ; Plasmids ; Rats ; Rats, Sprague-Dawley ; Surgical Flaps ; blood supply ; physiology ; Time Factors ; Transfection ; methods ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Objective: To evaluate the immunogenicity of HIV-1 clade C/B’ vaccine based on modified vaccinia virus Ankara (MVA) vector in mice. Methods: Mice were inoculated with 3-dose HIV vaccine by intramuscular injection. Blood sample were collected every second week, and then the antibodies against HIV were detected. At week 6, mice were killed and cellular immune responses were examined by ELISPOT. Result: The number of spot forming cells in the 107 pfu/ml -dose group was more than those of 105 pfu/ml -dose and 106 pfu/ml -dose groups significantly. HIV specific antibodies emerged at week 2 and elevated rapidly at week 4 and week 6. The level of specific IgG in the 107 pfu/ml -dose group was more than those of 105 pfu/ml -dose and 106 pfu/ml -dose groups significantly. Conclusion: The ADMVA induces both humoral immunoresponse and cellular immune responses.