Objective To investigate the effect of mixed-skin grafting with autologous microskin and allogenetic acellular dermal matrix microskin on wound healing in rats,and to make a further study on the related mechanism.Methods Wistar rats were served as a allogenetic acellular dermal matrix donor rats,and SD rats as acceptors with mould of full thickness skin defects on their back.The ninety SD rats were divided into 5 groups with 18 rats in each group.Group 1 was transplanted with autologous microskin,and group 2 with allogenetic acellular dermal matrix microskin.Groups 3,4 and 5 were grafted with mixed-skin ratio between autologous microskin and allogenetic acellular dermal matrix microskin 1 ∶ 1,1 ∶ 0.5 and 1 ∶ 0.25,repectively.The rate of wound healing was measured,wound samples collected,hematoxylin and eosin stain carried out,fibronectin (FN) and laminin (LN)detected,and intergroup comparison made,respectively,2,3 and 4 weeks after skin grafting.Results The wound healing rates and FN and LN expression of mixed-skin grafting groups were higher than those of the group with autologous microskin grafting.The group of 1 ∶ 0.25 obviously increased (P＜0.05 or P＜0.01).Conclusions The wound healing rate with mixed-skin grafting is higher than that with autologous microskin grafting.The best effect is achieved when the skin ratio between autologous microskin and allogenetic acellular dermal matrix microskin is 1 ∶ 0.25.It is possibly due to the increase of FN and LN on wound skin surface.

OBJECTIVETo evaluate therapeutic effect of the needle-knife closed solution combined with minor adjusting of spine for treatment of neck-shoulder syndrome.

METHODSFrom April 2010 to August 2011,120 patients with neck-shoulder syndrome were treated with the needle-knife closed solution combined with minor adjusting of spine, and included 45 males and 75 females and aged from 40 to 68 years old. The disease course was from 3 days to 10 years. After the operation, all patients taken the medicine of activating blood circulation herbs. At the 3rd, 7th, 10th day after operation, spinal rotation massage was performed on these patients. After the healing of the needle points, traditional Chinese medicine herb fumigation was applied on the needle points, and the patients were directed to do the cervical spine exercise. Therapeutic effect of the patients was evaluated by the neck disability index (NDI).

RESULTSAll patients were followed up after 3 weeks' treatment. The pain of neck-shoulder was relieved, and the range of motion was improved,the NDI score lowered from 49.30 +/- 1.35 before treatment to 10.15 +/- 1.18 at 3 weeks after treatment (t = 2.116, P < 0.05).

CONCLUSIONThe needle-knife closed solution combined with minor adjusting of spine for the treatment of neck-shoulder syndrome can relieve the pain in the neck-shoulder and improved the motion of the neck. The key for the effect is accurate location before operation, sufficient adhesion solution during the operation and spinal minor adjusting after operation.

Adult ; Aged ; Female ; Humans ; Male ; Manipulation, Spinal ; methods ; Middle Aged ; Myofascial Pain Syndromes ; therapy ; Neck Pain ; therapy ; Shoulder Pain ; therapy

Objective:To evaluate therapeutic effect of the needle knife closed solution combined with minor adjusting of spine for treatment of neck shoulder syndrome. Methods:From April 2010 to August 2011,120 patients with neck shoulder syndrome were treated with the needle knife closed solution combined with minor adjusting of spine ,and included 45 males and 75 females and aged from 40 to 68 years old. The disease course was from 3 days to 10 years. After the operation ,all pa-tients taken the medicine of activating blood circulation herbs. At the 3rd,7th,10th day after operation,spinal rotation massage was performed on these patients. After the healing of the needle points ,traditional Chinese medicine herb fumigation was ap-plied on the needle points,and the patients were directed to do the cervical spine exercise. Therapeutic effect of the patients was evaluated by the neck disability index (NDI). Results:All patients were followed up after 3 weeks'treatment. The pain of neck shoulder was relieved,and the range of motion was improved,the NDI score lowered from 49.30±1.35 before treatment to 10.15±1.18 at 3 weeks after treatment (t=2.116,P<0.05). Conclusion:The needle knife closed solution combined with minor adjusting of spine for the treatment of neck shoulder syndrome can relieve the pain in the neck shoulder and improved the mo-tion of the neck. The key for the effect is accurate location before operation ,sufficient adhesion solution during the operation and spinal minor adjusting after operation.

The aims of microbiology experiment teaching are not only to cultivate the students’ capacity of basic operation,but also to expanse their knowledge scope.We applied the bacterial diversity on the teach-ing class to make the students understand the progress of microorganism genomics research.It is helpful to cultivate the students’ innovative spirit and ability.Easy to work,clear result and low cost facilitated the spread of this experiment in the university.

Objective To establish a TaqMan real-time fluorescent quantitative PCR to detect Vibrio vulnificus based on hemolysin gene(vvhA)that coding cytolysin.Method By using software Primer Express, the PCR primers and TaqMan probe,which located in the conserved region of vvhA gene sequence,were designed for establishment of a TaqMan real-time fluorescent quantitative PCR to detect 100 bp amplicon from V.vulnificus DNA.A recombinant plasmid pMD19-vvhA100 as a positive control during detection was constructed using gene cloning technique.Minimal amplification cycles(Ct value)and fluorescence intensity enhancement (△Rn value)were used as observing index to optimize the reaction conditions of the TaqMan real-time fluorescent quantitative PCR.The DNAs with different concentrations from V.vulnificus and other eight bacteria and pMD19- vvhA100 were applied as templates to determine the specificity,sensitivity and reappearance of the TaqMan real- time fluorescent quantitative PCR.ICR mice were intraperitoneally,subcutaneously and orally infected with V. vulnificus,respectively.The detection effect of the TaqMan real-time fluorescent quantitative PCR was measured using the specimens of peripheral blood,subcutaneous tissue and intestinal content collected from the infected mice.Results The established TaqMan real-time fluorescent quantitative PCR showed positive results only for V. vulnificus DNA and pMD19-vvhA100.The detection effectiveness of the TaqMan real-time fluorescent quantitative PCR was as high as 0.01 ng of V.vulnificus DNA or 103 copies of pMD19-vvhA100.The SD values of the detection results repeated for three times using pMD19-vvhA100 with different concentrations were lease than 0.79. The detection results of TaqMan real-time fluorescent quantitative PCR were positive for all the specimens of peripheral blood and subcutaneous tissue.Conclusions The TaqMan real-time fluorescent quantitative PCR established in this study for V.vulnificus vvhA gene detection has advantages such as quickness,stability, sensitivity and specificity,indicating this method can be used for clinical laboratory diagnosis of septicemia and wound infection caused by V.vulnificus.

AIM: Tyrosinase gene was transfected into HEK293 cell as a reporter gene, it's property of synthesizing melanin, which can be examined by magnetic resonance imaging(MRI), is used to evaluate the tyrosinase gene's expression. The aim of this study was to search a way to evaluate the results of gene expression by MRI in vitro .METHODS: The plasmid of pcDNA3tyr which carried the full-length cDNA of tyrosinase gene was transfected into HEK293 cell by lipofectin. To observe the MRI signals of expressed melanin,the transfected cells were scanned by MRI sequences of T 1WI, T 1WI/SPIR and T 2WI. On the other hand, fontana stain was used to search for melanin granules in transfected cells, RT-PCR method was used to search for cDNA of tyrosinase gene. RESULTS: (1) Plasmids of pcDNA3tyr could be transfected into HEK293 cells and could synthesize a large amount of melanin. The synthetic melanins of 10 6 cells, which had been transfected 5?g, 10?g, 20?g plasmids of pcDNA3tyr separately, were all sufficient to be detected by MRI and appeared high signal in MRI T 1WI、T 1WI/SPIR、T 2WI sequences. The signal intensities of MRI imaging were related to the amounts of transfected plasmids positively. (2) The melanin granules could be found in HEK293 cells by Fontana stain. (3) The cDNA fragment of tyrosinase gene could be detected in transfected HEK293 cells by RT-PCR. CONCLUSION: The fact that MRI could detect the synthetic melanin of HEK293 cells, which controlled by expression of exogenous gene, demonstrates that medical imaging connecting with molecular biology technology can evaluate the result of gene expression in vitro .

Objective The purpose of our study was to investigate three dimensional dynamic contrast enhanced MR angiography(3D DCE M RA) in the detection of portosystemic collateral circulation in the falciform ligament in patients with portal hypertension. Methods From April 2003 to July 2008, 53 portal hypertension patients with varices in the falciform ligament were evaluated with 3D DCE MRA.Two radiologists independently assessed the number, diameter, location and drainages of the portosystemic collateral circulation in the falciform ligament according to the information on the 3D DCE MRA.Results The veins in the falciform ligament were classified into the superior and inferior groups, and both groups arise from the left trunk of the portal vein.In our study, the number of varices detected on 3D DCE MRA images varied from 1 to 3, and the diameters of these vessels varied from 0.4 to 2.6 cm.The inferior group consisted of paraumbilical/umbilical veins (47 cases), which flowed toward umbilicus and then drained upwards (n = 16) including deep superior epigastric veins (n = 7), superficial superior epigastric veins (n = 9), downwards (n = 40) including deep inferior epigastric veins (n = 7), superficial inferior epigastric veins (n = 33), or upwards and downwards at the same time (n = 9).The superior group of vessels in the falciform ligament were directly anastomosed with the internal thoracic vessels (n = 6).Conclusion In patients with portal hypertension, 3D DCE MRA can optimally demonstrate the portosystemic collateral circulation in the falciform ligament, which includes the superior and inferior drainage groups.

Objective To investigate the clinical value of three dimensional dynamic contrast enhanced MRA(3D DCE MRA)on esophageal and gastric varices compared with endoscopy.Methods From April 2003 to June 2008,153 patients with portal hypertension who underwent both 3D DCE MRA and endoscopy were reviewed retrospectively.All the patients were divided into bleeding group and non-bleeding group according to the clinical symptoms.The location and degree of the esophagogastric varices on 3D DCE MRA were assessed with postprocessing images,including subtraction,MIP and thin-slab maximum intensity projection(thin-MIP),and were compared with the results of endoscopy.The maximum,minimum and mean diameters of esophagogastric varices inside and outside of the wall were measured on the reformed images.The correlation between the findings of 3D DCE MRA and endoscopy were analyzed with Spearman rank correlation coefficient test The rates of esophagogastric varices outside of the wall in bleeding and non-bleeding group were compared by means of Chi-square test Results In bleeding group,severe esophageal varices were documented in 59 patients,moderate in 6 patients,mild in 5 patients;in non-bleeding group,severe esophageal varices were documented in 32 patients,moderate in 4 patients,mild in 5 patients.Severe,moderate,and mild gastric varices were documented in 28,34 and 16 in bleeding group,while they were 7,12 and 9 in non-bleeding group.Esophageal and gastric varices can be wholly presented on MIP images after subtraction,while the esophagogastric varices inside and outside of the wall can be differentiated on thin-MIP images.The location and degree of esophagogastric varices on 3D DCE MRA were correlated with the findings of the endoscopy.The range of r was from 0.544 to 0.878(P＜0.01).In 91 patients with severe esophageal varices,27 patients presented outside esophageal varices in bleeding group(n=59)and 24 patients presented outside esophageal varices in non-bleeding group(n=32).There was a significant difference in ratio of outside esophageal varices between bleeding group and nonbleeding group(X~2=7.199,P＜0.01).In 35 patients with severe gastric varices,22 patients showed adventitial gastric varices in bleeding group(n=28)and 4 patients showed adventitial gastric varices in non-bleeding group(n=7).The ratio of adventitial gastric varices in bleeding group was not significantly different from that of non-bleeding group(P=0.340).Conclusions 3D DCE MRA can display and differentiate the esophagogastric varices both inside and outside of the wall by three dimensional reconstruction.The results of 3D DCE MRA and endoscopy have good correlation.It is of importance in evaluating the esophageal varices outside of the wall,because they may indicate decreased risk of haemorrhage in patients with severe degree of esophageal varices.

Objective To probe into the clinical curative effect of laparoscopic high ligation of spermatic vein in the treatment of critical varicocele.Methods The clinical data of the 26 eases of laparoscopic high ligation of apermatic vein in the treatment of critical varicocde were reviewed and analyzed in the last two years in this hospi- tal.Results All the 26 cases had been conducted smoothly with the operation time 25～50min,an average of(28?3)min.After 3～24 months being followed-up,all the symptoms and signs disappeared with no relapse and testicle a- trophy.Eight wives of the patients who had been operated on got pregnant.Conclusion With soon recovery and small wound,it was safe to adopt laparoscopic high ligation of spermatic vein in the treatment of critical varicocele, especially for critical two-sided varicocde.

Objective To study the proliferation of in vitro cultured mouse chondrocytes exposed to different doses of fluoride.Methods The third generation of primary cultured chondrocytes were exposed to the concentrations of 0,5,10,20,40 mg/L fluoride for 10 days to observe the morphological changes under light microscope and electron microscope to counter the numbers of ehondrocytes and proliferating rote with the growth curve and MTT.Results After exposed to fluoride for 10 days,the proliferation was present in the chondrocytes of the 5,10,20 mg/L groups,and shrinked chromatine and apoptosed ehondrocytes were seen in 40 mg/L group.The absorbance was not significantly different between all groups(F=2.313,P＞0.05);after exposed to fluoride for 48 and 72 hours,there was a significant difference of proliferating ability among 0 mg/L(the contr01)group[(23.5±4.6)%,(29.9±1.7)%],5 mg/L group[(34.6±4.7)%,(45.3±5.9)%],10 mg/L group[(39.9±4.8)%.(56.8±5.5)%],20 mg/L group[(31.8±4.1)%,(38.3±6.5)%]and 40 mg/L group[(28.3±4.3)%,(33.4±4.8)%](F=11.401,25.671,P＜0.05).There wss a significant difference compared with the control group(P＜0.05)with that of 5 and 10 mg/L groups higher than that of 40 mg/L groups(P＜0.05).Conclusions Lower doses of fluoride improve the proliferation of in vitro mouse chondrocyte in a short exposing time,higher doses result in the opposite.