AIM: To quantitatively detect the expression level of transforming growth factor-β1 (TGF-β1) and transforming growth factor-β2 (TGF-β2) genes in the retina of normal rat in order to determine the expression difference of TGF-β1 and TGF-β2 in retina.METHODS: The total RNA was isolated from which the first strand of cDNA was prepared. The mRNA levels of TGF-β1 and TGF-β2 were detected quantitatively by real time polymerase chain reaction (PCR).RESULTS: The mRNA levels of TGF-β1 and TGF-β2 were 0.0008±0.0003 and 0.0378±0.009, respectively. Expression of TGF-β2 was obviously higher than that of TGF-β1 in rat retina with statistical significance (t=12.37, P<0.001). The ratio of TGF-β2/TG-β1 was 55.00±26.61.CONCLUSION: QRT-PCR could specifically and accurately detect gene expression level in rat retina. In retina the TGF-β2 gene was expressed more abundantly than TGF-β1. It is suggested that TGF-β2 play an important role in retina diseases.

AIM: To quantitatively investigate transforming growth factor-β type Ⅰ receptor (TβRⅠ) and transforming growth factor-β type Ⅱ receptor (TβRⅡ) gene expressions in rat retina.METHODS: Sprague-Dawley rats were chosen in this research. Gene expression was detected quantitatively by reverse transcription polymerase chain reaction (RT-PCR) analysis. RESULTS: The expression level of TβRⅠ and TβRⅡ were 0.00034±0.00013 and 0.0001±0.00005, respectively. The expression level of TβRⅠ was obviously higher than that of TβRⅡ in the rat retina with statistical significance (P<0.01). The ratio of TβRⅠ/TβRⅡ was 3.9±1.7.CONCLUSION: Real time quantitative RT-PCR is an effective method to detect differential expression genes in retina. The change of TβRⅠ/TβRⅡ expression may play an important role in the pathogenesis of retinopathy, which could be further investigated in its significance in the development of proliferation retinopathy.

AIM: To detect the gene expression of TGF-β2 in retinas of diabetic rats at different stages, to observe and analyze the effect of TGF-β2 on the retinas of diabetic rats, to explore the role of TGF-β2 in pathogenesis of diabetic retinopathy (DR), and to provide experiment data and experience for further clinic studies.METHODS: Sprague-Dawley (SD) rats were used and retinas were dissected. The total RNA was isolated from which the first strand of cDNA was prepared. Diabetes mellitus was induced by a single intraperitoneal injection of 60mg/kg streptozotocin (STZ) and the rats were held without insulin treatment until sacrifice. Besides, agematched rats treated with saline were used as controls. Tail vein blood glucose was measured after 2 days and rats were considered hyperglycemic if blood glucose reading>16.7mmol/L. Animals with blood glucose level<16.7mmol/L were excluded from the study. The rats were killed at the 4th, 8th, 12th, 16th, 20th and 24th week respectively after hyperglycemic models were established. The retinas were separated and preserved in liquid nitrogen. The expressions of TGF-β2 gene mRNA were detected by reverse transcription PCR (RT-PCR).RESULTS: The RNA of rat retina was integrative enough to be used to further carry out PCR analysis. Compared with control groups, the expression of TGF-β2 mRNA in retinas of diabetic rats was up-regulated at the 4th week, but there was no statistical difference (P>0.05); it was down-regulated at the 8th week, and there was statistical difference (P<0.05); it was also down-regulated at the 12th week, and there was statistical difference (P<0.05); at the 16th week there was no statistical difference (P>0.05); it was up-regulated at the 20th week, but there was no statistical difference (P>0.05); it continued to be up-regulated at the 24th week, and there was statistical difference (P<0.05).CONCLUSION: Since the expression of TGF-β2 mRNA in retinas of diabetic rats was down-regulated at the 8th week and 12th week statistically, up-regulated at the 24th week statistically, it has obviously shown that TGF-β2 was down-and up-regulated through the period of DR. That is, its changes are diphasic with time. It may confirm that TGF-β2, with the characteristic of diphasic regulation, played an important role in DR. It is necessary to study it furthermore.

Objective To explore the efficient way to purify and to enrich umbilical cord mesenchymal stem cells which have abilities to differentiate into chondrocytes. Methods Umbilical cord was acquired from new born infants and chopped into 1 mm2 cubics, then they were pretreated with colleganase and trypsin so as to get the mixed cell suspension. CD45(-),CD90(+) cells were acquired from the mixed cell suspension via FCM sorting and were cultured in the medium as umbilical cord mesenchymal stem PO cells. The PO cells were detached and passaged by multiplex collagenase NB4 and the positive rate of CD90 and CD45 of the P1-P3 cells were observeed via FCM ; the P3 cells were induced to differentiated into chondrocytes. Results FCM results showed that the positive rate of CD90 on P1, P2, F3 was (80.86 ± 7.85)%,(95.86± 3.28)%,(97.15± 1.43)%, while the positive rate of CD45 on P1, P2, P3 cells was (2.53 ±0.28)% and (0.97 ± 0.48)% and (0.05 ± 0.01)% respectively; P3 cells had differenciated into chondrocytes. Conclusion FCM presorting and Multiplex collagenase detachment procedures can swiftly acquire highly purified human umbilical cord mesenchymal stem cells which have the ability to differentiate into chondroeytes.

ObjectiveTo observe the effection of different interventions of group B streptococcus (GBS) infection in gestation period.MethodsOne hundred and seventeen cases with GBS infection were obtained from 1885 pregnant women,who were got routine prenatal examination at 34 to 37 weeks of pregnancy,and drug seeitivity test of secretions which were taken from under paragraph 1/3 of vagina,and divided into treatment group (91 cases) and untreatment group (26 cases).The treatment group was divided into treatment group one (47 cases) and treatment group two (44 cases).Treatment group one was treated with oral antibiotics for 7 days after diagnosis,treatment group one and two were treated with postpartum antibiotics intravenous infusion once every 4 hours in labor.Comparison of maternal and fetal outcomes.ResultsThe GBS infection rate was 6.2％ (117/1885).The morbidity of premature delivery,premature rupture of membrane and neonatal infection of treatment group [ 5.5％ ( 5/91 ),13.2％ ( 12/91 ),5.5％ (5/91 ) ]were lower than those of untreatment group [ 19.2％(5/26),30.8％(8/26),23.1％(6/26) ](P ＜ 0.05).The morbidity of premature dehvery and premature rupture of membrane of treatment group one[ 0,6.4％ (3/47)]were lower than those of treatment group two[ 11.4％ (5/44),20.5％ (9/44)](P ＜ 0.05).Conclusion Anti-GBS treatment can improve the outcomes of mothers and infants,especially early anti-GBS treatment during the period of pregnancy.

· AIM: To quantitatively detect gene expression level of transforming growth factor-β type 1 receptor (Tβ R 1) and transforming growth factor-β type 2 receptor (Tβ R 2) in different stage of diabetic rats' retina. To observe and analyze the effect of transforming growth factor-βreceptors on the retina of rat diabetic animal model.· METHODS: 28 healthy adult Sprague-Dawley rats were chosen and randomly divided into two groups of normal control (CON) and diabetes mellitus (DM). Diabetes was induced by streptozotocin (STZ) intraperitoneal injection.Gene expression was detected quantitatively with real-time fluorescence quantitative reverse transcription polymerase chain reaction (QRT-PCR).· RESULTS: The mRNA level of Tβ R 1 and Tβ R 2 was 0.000493± 0.000133 and 0.000166± 0.000057 at 4wk. The mRNA level of Tβ R 1 and Tβ R 2 was 0.000608± 0.000232 and 0.000113± 0.000049 at 12wk. Tβ R 1 expression was gradually elevated during the progression of diabetic retinopathy, Tβ R 2 expression was up-regulated at 4wk,but down-regulated at 12wk.Tβ R 2) may play important role in the pathogenesis of diabetic retinopathy.

Objective To develop a liquid chromatography-tandem mass spectrometry (LC-MS/MS) screen-ing method for 45 poisonous alkaloids in blood. Methods Identification was based on the compound’s retention time and two precursor-to-production transitions. The method involved a liquid-liquid extraction (LLE) followed by LC-MS/MS with multiple-reaction monitoring (MRM). When 1 mL of blood was ex-tracted with diethyl ether at pH=9.2 with SKF525A as the internal standard, the target compounds were an-alyzed with LC-MS/MS in the positive ionization mode. Results The target alkaloids had good linearity (r>0.995 1), both the intra-day precision and inter-day precision being less than 14.77%. The limits of detection ranged from 0.05 to 25 ng/mL in blood. Conclusion The method is selective and sensitive in detecting poisonous alkaloids with a total running time of 12 minutes; therefore it was successfully ap-plied to some actual cases of suspected alkaloids poisoning.

Objective To investigate the effects of sulfur dioxide(SO2) pollution in different concentrations on activity of SOD,levels of GSH and MDA in heart and lung tissue of exercised rats,in order to elucidate the toxicological mechanism of SO2 pollution.Methods Forty-eight SD rats were randomly divided into eight groups:a rest group(RG),an exercise group(EG),a low pollution exercise group(LEG),a low pollution rest group(LRG),a moderate pollution exercise group(MEG),a moderate pollution rest group(MRG),a high pollution exercise group(HEG) and a high pollution rest group(HRG).All groups,except RG and EG,were exposed to SO2 with different concentrations(5 mg/m3,10 mg/m3,15 mg/m3),meanwhile exercised rats run in a motor-driven wheel at a speed of 8 m/min,2 h/d for 10 days.Rats were sacrificed at 24 h after treatment and the anti-oxidative enzymes activity and lipid peroxidation(LPO) levels were measured.Results Compared with RG,SOD activity of lung tissue significantly decreased in HEG,MEG,LEG(P

Objective To evaluate the effect of ImmunoCyt on the early diagnosis of bladder cancer by comparing the result of cystoscopy with that of cytology. Methods Eighty-six consecutive patients with bladder cancer (mean age,63 years) were included in this group.Among them 38 had undergone transurethral resection of superficial transitional cell carcinoma and had been followed up and 48 had hematuria.These patients were evaluated with standard cytology and ImmunoCyt test.Then all the patients underwent cystoscopy and suspected lesions were biopsied. Results Transitional cell carcinoma of bladder was confirmed pathologically in 28 cases.The sensitivity of ImmunoCyt (82%) was superior to that of cytology (39%, P

Objective To review the clinical and laboratory characteristics of the hemohemocytopenia diseases in children ,in or-der to improve the understanding of the related diseases and the level of diagnosis and treatment .Methods A total of 217 children with hemocytopenia diseases with one ,two or three series of cell reducing ,and the degree of blood cells reduce with anemia and common diseases in children .Results In 185 cases with the hemocytopenia diseases ,1 series reduce were mainly in idiopathic thrombocytopenic purpura(ITP ,37 .2% )、thrombocytopenia (19 .8% ) ,granulocytopenia or agranulocytosis (9 .1% ) .2 series re-duces mainly in acute leukemia (43 .6% ) ,the IT P (21 .7% ) and aplastic anemia (6 .5% ) .3 series reduce mainly in acute leukemia (38 .9% ) and aplastic anemia(33 .6% ) .Common cause of anemia in children with 180 cases of hemohemocytopenia diseases were a-cute leukemia (20 .0% ) ,the ITP(19 .4% ) and no clinical diagnosis of anemia (16 .1% ) .Conclusion After analyzing the character-istics of the patient′s clinical and laboratory in 217 cases of hemocytopenia diseases ,improve the clinical blood disease related knowl-edge ,help understand the laboratory data and proportion .