BACKGROUND:The low output of seed cells and long cycle of traditional ceils culture methods in tendon animal models(rabbits and chicken) restrict the researches of tendon tissue engineering study.OBJECTIVE:To establish an ideal culture protocol of tail tendon in SD rats,to get more seed cells within less time for subsequent engineered tendon construction research.DESIGN,TIME AND SETTING:Controlled observation was performed in the National Key Laboratory of Biotherapy,Department of Stem Cells and Tissue Engineering,Sichuan University between February and November in 2006.MATERIALS:Rat tail tendon cells were harvested from 2 SD rats,aged 7-10 days;human prepuce fibroblasts were offered by National Key Laboratory of Biotherapy,Department of Stem Cells and Tissue Engineering,Sichuan University.METHODS:Tail tendon of SD rats was draw off and cut into pieces,which were then cultured in 10% fetal bovine serum+DFculture medium for getting primary tendoncyte by using suspension tissue culture method. The third generation cells were processed into immuocytochemistry stain with collagen type Ⅰ and Ⅲ,while human prepuce fibroblasts served as controls.Absorbance of stain result was measured by image-pro plus 5.02 for statistical analysis.MAIN OUTCOME MEASURES:Immuocytochemistry stain and absorbance measurement of SD rat tail tendon cells.RESULTS:The second generation of SD rat tail tendon cells were positive for type Ⅰ collagen stain,and negative for type Ⅲ collagen stain;human fibroblast were positive for both Ⅰ and Ⅲ collagen. In the rat tail tendon cells and human flbroblasts,the absorbance value of type Ⅰ collagen expression was dramatically higher than of type Ⅲ collagen(P＜0.05). There was no significant differences addressing the absorbance of type Ⅲ collagen expression between type Ⅰ and Ⅲ collagen of SD rat tail tendon cells and blank control group (P＞0.05).CONCLUSION:Cells cultured from SD rat tail tendon have biological characteristic of tendon cells. Tissue piece suspensionculture can obtain a quantity of primary or subcuitured cells of rat tail tendon within a short time.

Objective To clone human vacuolar protein sorting 4A gene(hVPS4A)and to construct its eukaryotic expressive plasmid.Methods Primers were designed to amplify the full length hVPS4A by PCR using cDNA of Huh7 cell as a template,then the target DNA was inserted into the eukaryotic vector pRK5.The recombinant plasmid was confirmed by PCR,restriction enzyme digestion and DNA sequencing.Results A 1 300 bp fragment was successfully amplified by PCR from the cDNA of Huh7 cells.Af-ter recycled,purified and ligated with the vector pRK5,the recombinant plasmid was transformed into E.coli DH5α.The positive re-combinant plasmid identified by PCR was selectred and digested by EcoRⅠto get a 5 900 bp fragment;and two fragments including 4 600 bp and 1 350 bp were obtained using EcoRⅠand HindⅢ digestion;the size of these two fragments were consistent with the pRK5 target fragment and the inserted hVPS4A as expected.Moreover,DNA sequencing results confirmed that the inserted frag-ment was in accordance with the hVPS4A reference sequence.Conclusion The eukaryotic expression vector containing hVPS4A gene is constructed successfully,which provides the condition for further study on the hVPS4A biological functions.

Ninety rats were divided into 8 groups. Group 1-5 were fed vitamin A free ration and each group was divided into 2 subgroups at the beginning and 12, 23, 36, 43, days after the experiment respectively. Group 6-8 were fed the ration containing 66, 132, and 198?g vitamin A per 100g diet respectively fer 30 days, and each group was also divided into 2 subgroups. The results showed that RBPR% was better than the serum vitamin A content in evaluating vitamin A nutritional status. It was suggested that the vitamin A nutritional status could be graded as follows: deficient(abore 70%)) mar-ginal(60-70%),normal (20-60%), and sufficient (under 20%)by RBPR%.

An improved microfluorimetric method for determination of serum vitamin A was presented. In this method, the excitation wavelength 345nm and emission maximum 480nm were selected, and cyclohexane as solvent for extraction of saponified vitamin A in the serum samples to be tested was used. The sensitivity was sufficient for measurement of vitamin A content in 20*l serum taken from normal fasting blood without elimination of interference. The recovery averaged 99.0% (93.0-105.5).When this method was applied to laboratory study or field work, practically, the results showed that the values of vitamin A obtained seemed to correlate with its nutritional status of experimental animals or subjects investigated. The procedure was simple, reproducible and easily acceptable, espicially in the field work.

35 rats were divided into 5 groups. Group 1-3 were fed rations containing 20, 12 and 6% caseien respectively, group 4 containing 12% mixed protein of soybean and gluten. The food of group 5 was the same as that of group 1 but the amount was only half of that of group 1. Every animal was given 8?g vitamin A by stomach tube daily for 3 weeks and then 25?g for 2 weeks. Afterward serum vitamin A values were determined in all animals, then 132?g vitamin A was administrated by stomach tube. After 3.5 hours, the vitamin A values in serum and liver were determind. The results showed that in group 3 and 5 serum vitamin A contents decreased and the liver values rose. It was suggested that the release of vitamin A from liver had been interfered by semistarvation or low protein intake. There is no difference between group 2 and 4.

The ingredients of five kinds of Zhejiang's yellow Chrysanthemum morifolium with different flower blossoming stages were comparatively analyzed. Polysaccharides, total flavonoids, volatile oil, alcohol extract, water extract, chlorogenic acid, luteolin, 3,5-O-dicaffeoyl quinic acid and fingerprint of the ingredient were determined as indicators. During flower blossoming stages, the ingredients of Ch. morifolium showed a big difference with a certain variation. At the early opening stage, the contents of flavonoids and volatile oil were higher, the content of chlorogenic acid, luteolin, 3,5-O-dicaffeoyl quinic acid were higher in the middle of the flowers 50% -80% fowers blossoming degree is the optimal time for harvest.
China ; Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; Chrysanthemum ; chemistry ; growth & development ; Drugs, Chinese Herbal ; analysis ; Flavonoids ; analysis ; Flowers ; chemistry ; growth & development ; Quality Control ; Quinic Acid ; analysis

Objective To study the clinical characteristics and curative effects of pancreatic cystade- nocarcinoma in order to improve its diagnostic and therapeutic accuracy.Methods A retrospective analysis was done on the clinical materials of 13 cases of pancreatic cystadenocarcinoma hospitalized in Shanxi Cancer Hospital from 1990 to 2006.Results The preoperative diagnosis were as follows:pancreatic cystadenocarci- noma 6 cases,pancreatic cystadenoma 2 cases,pancreatic cancer 1 case,pancreatic pseudocyst 4 cases.The misdiagnosis rate was 53.8 %.Surgical operation was done on the 13 cases,and 10 of them were treated by radical operation.A 5-year follow-up was done on 6 still alive cases,and 1 of them lived over 11 years.3 cases were treated by palliative operation,and all of them died within 3 years.Conclusion Since there is no specific clinical manifestations of pancreatic cystadenocarcinoma,it is very difficult to get an accurate preop- erative diagnosis.Radical operation is the most effective therapeutic methods.

Background Researches demonstrated that ciliary neurotrophic factor (CNTF) can enhance survival and promote differentiation of neutron.Meanwhile,CNTF also is thought to play an important role in the development of visual pathway.But,less studies are reported in the relationship of CNTF and form deprivation amblyopia.Objective This study was to investigate the expressions of CNTF in visual cortex area 17 in form deprivation amblyopia model.Methods Twelve 4-week-old cats were randomized into normal group and form deprivation amblyopia group.Monocular form deprivation amblyopic models were established in 6 cats by eyelids suture method.Pattern visual evoked potential(P-VEP) was recorded to evaluate the amblyopic models 16 weeks later following the eyelids suturing.Then,bilateral visual cortex tissue was incised at a vertical in sagittal axis fashion to prepare the section.Nissl staining was used to detect the morphologies of neurons.Expression of CNTF in Ⅰ-Ⅵ layers of visual cortex area 17 was located and quantified by immunochemistry.The positive cell number and gray scale for CNTF were calculated and compared between two groups.The use of the animals complied with Regulations for the Administration of Affairs Coucerning Experimental Animals by State Science and Technology Commission.Results Compared with the normal group,P-VEP amplitude was significantly reduced (6.11 ±1.56 μV vs.11.42±t.92 μV) and latency was significantly prolonged(75.77±9.83 ms vs.58.56±7.17 ms) in the form deprivation amblyopia group (t=5.272,3.464,P＜0.05).Nissl staining showed that the number of neurons in the form deprivation amblyopia group was less than that in the normal group.In the form deprivation amblyopia group,neurons became shrinkage and turned round,cytoplasmic processes get shortened,and the nucleus became small.The number of Nissl bodies was decreased.lmmunochemistry showed the positive neutrons for CNTF in Ⅰ-Ⅵ layers of visual cortex area 17 in hoth normal cats and model cats with the more positive cells in Ⅱ-Ⅳ layers.Compared with the normal group,the positive cell number for CNTF was significantly reduced in Ⅱ-Ⅳ layers of visual cortex area 17 in the form deprivation amblyopia group (Ⅱ layer:95.93±8.24 vs.116.25±6.52;I layer:102.65±7.45 vs.125.23±8.21;Ⅳ layer:l10.65±6.85 vs.139.54±4.26) (t=4.737,4.989,8.773,P＜0.05).In addition,the gray scale of CNTF positive cells was significantly lower in Ⅱ-Ⅳ layers of visual cortex area 17 in the form deprivation amblyopia group than that the normal group (Ⅱ layer:106.98 ± 8.86 vs.138.65 ± 6.38 ; Ⅲ layer:109.56 ± 8.69 vs.149.59 ±8.55;Ⅳ layer:l16.65 ±9.52 vs.155.76±9.87) (t=7.105,8.043,6.986,P＜0.05).Both CNTF positive cell number and gray scale in Ⅰ,Ⅴ,Ⅵ layers of visual cortex area 17 had no significant differences between two groups (P＞0.05).Conclusions Form deprivation in critical period of a new born animal may lead to distributing abnormality of CNTF in visual cortex,which maybe play a role in the development of form deprived amblyopia.

AIM: To screen TIGR/myocilin gene (MYOC) mutation in high myopic Chinese children with family history.METHODS: Gene sequencing was performed in exon 3 of the TIGR gene in high myopic Chinese Children. The coding sequence of TIGR exon 3 was screened by capillary electrophoresis sequencing. The sequence alterations were analyzed by bioinformatics.RESULTS: TIGR gene mutation was not found in high myopic patients and normal controls group.CONCLUSION: No identified gene mutation is found in TIGR gene in high myopic Chinese children.

Objective To study the association between clinical symptoms,psychosomatic factors and autonomic nervous function in patients with gastroesophageal reflux disease (GERD).Methods Thirty-four patients with GERD diagnosed by reflux disease questionnaire (RDQ) and endoscopy and 15 healthy control subjects were enrolled in this study.All the subjects were divided into two groups,one with normal scores of Zung's self-rating anxiety scale (SAS) and Zung's depression scale (SDS) as GERD (-) and the other with abnormal scores of SAS and SDS as GERD (+).Reflux symptom score was recorded for both groups at the same time.Autonomic nervous function was assessed by their heart rate variability (HRV) on dynamic electrocardiogram (DCG).The time domain parameters analyzed included standard deviation (SD) of average R-R interval during 24 hours (SDNN),SD of average 5-minute sinus heart rate (SDANN),mean square root of the difference of adjoining two R-R interval (rMSSD),and proportion of the heart beats with difference of R-R interval more than 50 ms from the total heart beats (PNN 50),and the frequency domain parameters analyzed included low-frequency (LF),high-frequency (HF) and ratio of LF to HF.Results Average scores of SAS and SDS were significantly higher in patients with GERD than those in healthy controls (48?9 vs 38?6 and 48?11 vs 41?6,respectively,P