Objective To evaluate the possibility and efficiency of nanoparticles as a new vector in vascular endothelial growth factor (VEGF) gene transfection. Methods Nanoparticle-VEGF (Np/VEGF)complex was prepared with poly (D, L-lactide-co-glycolide) (PLGA) loading VEGF165 gene using the multiple emulsion (w/o/w) technique. The envelopment efficiency and size of the complex were determined. Rat myocardial cells were cultured in vitro, and the Np/VEGF was transfected into the cultured myocardial cells. Then RT-PCR and ELISA were used to evaluate whether the Np/VEGF increased the level of gene expression. Four New Zealand rabbits were used, the suspension of Np/VEGF was injected into myocardial tissue of rabbits after thoracotomy. 96h after the operation, the tissue sections of the implant sites were observed with transmission electron microscope (TEM) to determine the process of nanoparticles as vectors for gene transfer to cardiac myocytes. Results The envelopment efficiency and size of the Np/VEGF complex thus prepared were 1.87% and 25-300nm respectively. RT-PCR and ELISA showed that VEGF gene could be successfully transfected into myocardial cells by nanoparticle, and NP/VEGF significantly enhanced gene transfection efficiency, and it was more effective than plasmid. 96h after the operation, a great number of nanoparticles were observed in myocardial cytoplasm and nucleus with TEM, and many nanoparticles began to dissolve and degrade, suggesting that the DNA was released slowly from the nanoparticles localized in the cytoplasmic compartment, and was then transferred into the nucleus. Conclusions NP/VEGF can act as a vector to transfect VEGF gene in vitro and in vivo, it significantly enhanced gene transfection efficiency, and it was more effective than plasmid.

Objective To evaluate the clinical results using modified Richards nail treating femoral intertrochanteric comminuted fractures.Methods From March 2002 to March 2005,69 patients suffered from femoral intertrochanteric comminuted fractures belong to Evan Ⅲ type or Ⅳ type were randomly divided into experimental group and control group.The experimental group was treated with modified Richards nail,including 34 patients,18 males and 16 females.The patients were from 61 to 78 years old,and the mean age was 67.5 years old.In this group,21 patients were involved in left femoral intertrochanteric comminuted fractures,13 patients were involved in right.The pattern of the fractures included EvanⅢ type in 24 cases and Ⅳ type in 10 cases.The control group including 35 patients was treated with DHS,and there were 19 males and 16 females with an average age of 65.9 years(ranged 61 to 80 years).In this group,19 patients were involved in left femoral intertrochanteric comminuted fractures,16 patients were involved in right.The pattern of the fractures included EvanⅢ type in 25 cases and Ⅳ type in 10 cases.Results The experimental group were followed-up 9-23 months with a mean of 11.60?3.78 months.The control group were followed-up 9-24 months with a mean of 11.40?4.12 months.In experimental group,the average fracture healing time was 3.65?0.97 months.While in the control group,the average fracture healing time was 4.32?1.38 months.In experimental group,the leg length measured at the 9th month post-operatively was averagely(0.82?0.36)cm shorter in the diseased side than in the healthy side,while in control group the leg length was averagely(1.08?0.51)cm shorter.There was significant difference between two groups.Compared to the control group,the curative effect was better in the experimental group.Conclusion Treating femoral intertrochanteric comminuted fractures with the modified Richards nail shows a good result,which can effectively diminish the non-union or mal-union of the fractures.

Acupuncture as an effective complementary and alternative therapy can effectively relieve inflammatory and neurogenic pain, and a proper animal model is a key link to ensure the quality of research in acupuncture analgesia. This article makes some suggestions for improving model making methods, unifying model assessing criteria, and selecting and locating acupoints by analyzing present commonly used animal models of acupuncture analgesia and the present situation of their application and identifying some problems about model making methods, evaluation indices, acupoint selection and clinical transformation existing in animal models of acupuncture analgesia in order to promote the development of basic research in acupuncture.

Objective To establish a real‐time quantitative PCR method for the detection of cytokine receptor‐like factor 2 (CRLF2) expression .Methods Specific primers amplification target gene CRLF2 and housekeeping genes ABL were designed ,the purified PCR products were performed the TA cloning .After bacterial colony PCR screening and sequencing ,then the recombinant plasmids DNA was extracted and measured by using UV spectrophotometer and converted to copies/mL concentration .Finally it was diluted for preparing the plasmid standard substance ,then the standard curve was drawn for observing the sensitivity and linear rang ,meanwhile the stability of the plasmid DNA was evaluated .This method was initially applied to detect the CRLF2 level of bone marrow mononuclear cells in 10 cases of healthy children and 10 cases of newly diagnosed acute lymphoblastic leukemia (ALL) .Results CRLF2 PCR product had a single specific melting curve;the linear detection range of the standard substance was 103 - 108 copies /ml;the plasmid standard substance by freeze‐thawing for 3 times remained stable;the CRLF2 level of clinical sample was within the linear detection range of standard substance .Conclusion The real‐time quantitative PCR method for CRLF2 established by our laboratory has good specificity ,linearity range and stability ,which can be applied to the quantitative detection of CRLF2 gene in clinical ALL children .

AIM:To study the differences of basic fibroblast growth factor(bFGF)gene expression in lens epithelial cells (LECs) between fetuses and cataract patients. METHODS: In situ hybridization was used to detect bFGF mRNA in the LECs that were cultured and in tissue sections from fetuses and in the LECs from the anterior capsule of cataract patients. Image analysis was used for the relative quantitative analysis of bFGF mRNA. RESULTS: bFGF gene existed in the LECs that were cultured and in tissue sections from fetuses and in the LECs from the anterior capsule of cataract patients. The integral absorbance for the fetal cultured cells, the fetal tissue sections and the capsule membrane cells of cataract patients were 627.1±268.7, 131.5±42.8 and 79.2±26.3 respectively. The integral absorbance of fetal cultured LECs was significantly higher than that of fetal section LECs (P<0.01). The integral absorbance of cataract LECs was significantly lower than that of fetal LECs (P<0.01). CONCLUSION: The in vitro culture of LECs can improve bFGF gene expression. The bFGF gene expression in fetal LECs is significantly higher than that in cataract LECs.

Catheter Ablation ; methods ; Humans ; Hypertension ; surgery ; Renal Artery ; Sympathectomy ; methods

Objective:To investigate the expression of wild type estrogen receptor(wER)and the ex-on-5 deleted ER(variant ER,vER)in human hepatocellular carcinoma(HCC)samples,and thereafteranalyze the possibility of HCC treatment by endocrine therapy.Methods:The mRNA expressions of wERand vER were analysed from 28 cases of HCC by RT-PCR.The expression of ER at the protein level wasdetected by immunohistochemistry(IHC).Results:IHC results showed that 39.3% of the HCC speci-mens expressed ER.The mRNA of wER was detected in 89.3%(25/28)of the HCC specimens whilethat of vER was detected in 96.4%(27/28).Twenty four out of 28 HCC cases(85.7%)expressedboth wER and vER.One out of 28 patients(3.5%)expressed only wER whereas 3 patients out of 28(10.7%)expressed vER only.Conclusion:Ninety six percent(27/28)of the HCC patients expressedvER,which suggests that the expression of vER is an important event in the development of HCC.

Acupuncture Therapy ; Adult ; Aged ; Chronic Disease ; therapy ; Female ; Humans ; Male ; Middle Aged ; Pharyngitis ; therapy ; Young Adult

OBJECTIVE To define the regularity of survival ability of HIV in natural environment,and prevent(infection) through contacting with positive body fluids during daily life or medical work.METHODS Having been diluted by sterile water or 10% serum RPMI 1640 medium,HIV was exposed to 4℃,room temperature(20-26℃) or 37℃ for different period of time.TCID_(50) of these samples was detected.Non-pathological samples were blind passaged for three generations.RESULTS HIV infective ability persisted more than 35 days both in(water) and medium at 4℃;whereas it persisted 7-14 days in water,14-21 days in medium at room temperature and 37℃.CONCLUSIONS HIV has higher resistance in natural environment.To prevent accidental spreading of HIV,HIV positive liquids and contaminants staffs should be treated carefully.

Objective To study the efficacy and safety of tacalcitol combined with monochromatic excimer light (MEL) 308 nm vs MEL 308 nm monotherapy in treating vitiligo.Methods Thirty-eight pa- tients with vitiligo were enrolled in the single-blind clinical trial,using plabebo-treated lesions in the same patient as controls.Contralateral or nearby lesions were randomly selected to be treated by either tacalcitol or placebo.All lesions were treated weekly with MEL 308 nm,for a total of 12 sessions.Patients were ex- amined at monthly intervals.The mean number of sessions and the cumulative dosage for initial and excel- lent repigrnentation were calculated.Results Thirty-five patients were evaluated.The mean?SEM cumu- lative dose and number of MEL exposures for initial repigmentation,respectively,were 4.27?3.59 J/cm~2 and 4.89?3.16 on tacalcitol-treated site,5.36?4.12 J/cm~2 and 5.69?3.29 on placebo-treated site,re- spectively (both P＜0.05).For excellent repigrnentation,the cumulative dose and number of exposures were 7.72?5.64 J/cm~2 and 7.79?4.70 respectively on tacalcitol-treated site,and 8.18?4.87 J/cm~2 and 8.4?3.92 respectively on placebo-treated site (both P＞0.05).Treatment with tacalcitol resulted in a sig- nificantly higher percentage (71.4% vs 54.3%) of repigmentation than that with placebo.Conclusions Our results show that MEL 308 nm is safe and effective for the treatment of vitiligo.Additionally,concur- rent topical tacalcitol potentiates the efficacy of MEL 308 nm in the treatment of vitiligo;this combination achieves more rapid pigmentation with a lower total MEL dosage.