Abundant experiences have already been accumulated in treatment of stroke with acupuncture. Development of rehabilitation theory also brings unprecedented opportunity and challenge to acupuncture. Combined with the modern rehabilitation theory and practice, it is very helpful to deepen the understanding on treatment of acupuncture for cerebral apoplexy and enhance the therapeutic effect in clinic by studying the mechanism of acupuncture treatment, opportunity of intervention, selection of acupoints, needling manipulations and quantity of stimulations etc. Through analysis on the necessity and the way of combination of acupuncture and modern rehabilitation, it is concluded that rehabilitation evaluation, rehabilitation phases and obstacle analysis should be taken as references by acupuncturists to reinforce the therapeutic effect and creditability of acupuncture treatment on cerebral apoplexy.
Acupuncture Therapy ; methods ; Hemiplegia ; rehabilitation ; therapy ; Humans ; Stroke ; therapy ; Stroke Rehabilitation

Objective To establish a rapid, sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of acyclovir (the metabolite of valacyclovir hydrochloride) in human plasma. Methods After addition of ganciclovir as internal standard (IS), plasma samples were prepared by one-step protein precipitation using acetonitrile as precipitant, followed by an isocratic elution with 0.1% formic acid 3.5μm) column. Detection was performed on a triple-quadrupole mass spectrometer utilizing electrospray ionization (ESI) interface operating in positive ion and selected reaction monitoring (SRM) mode with the precursor to product ion transitions m/z 226.2→152.1 for acyclovir and m/z 256.2→152.1 for the IS. Results The analytical results demonstrated a good linearity over the ranges from 0.005 to 4μg/mL (r=0.9999) for valacyclovir hydrochloride. The relative standard deviations (RSD) of intra-batch and inter-batch were less than 4.06% and 9.23%, respectively. The limit of detection and lower limit of quantification in human plasma were 2ng/mL and 5ng/mL, respectively. Conclusion The method was simple, sensitive, accurate and reproducible and has been successfully applied to a bioequivalence study of valacyclovir hydrochloride capsules in Chinese healthy male volunteers.

Objective To study the value of the diameters of fetal umbilical artery and umbilical vein in early prenatal screening program for thalassemia by ultrasound.Methods A total of 120 cases of singleton pregnant with thalassemia at 12- 28 gestational weeks were detected by ultrasound,328 cases of normal singleton pregnant at 12 - 28 gestational weeks were selected as controls.The diameters of fetal umbilical artery and umbilical vein were tested and compared between the two groups.The diagnostic values were evaluated by ROC curve.Results At 12 - 15+6 gestational weeks,the diameters of umbilical artery and umbilical vein of thalassemia group were larger than those of control group(P ＜0.05).At 16 - 19+6 gestational weeks, the diameter of umbilical vein of thalassemia group was larger than that of control group,the differences were significant (P ＜0.05).The cutoff threshold of the diameter of fetal umbilical artery was 1.80 mm,the diameter of fetal umbilical vein was 2.95 mm at 12 - 15+6 gestational weeks,the diameter of fetal umbilical vein was 3.80 mm at 16- 19+6 gestational weeks,which were obtained by ROC analysis.The sensitivity of this cutoff threshold to identify thalassemia were 83.3％, 83.3％, 47.8％,respectively, while the specificity were 90.5 ％, 95.2 ％, 85.4％, respectively.Conclusions The diameters of fetal umbilical artery and umbilical vein detected by ultrasound have great value on early prenatal screening of thalassemia, these indexes can be used as an important factors in early prenatal screening of thalassemia.

Background To optimize the culture method of human retinal microvascular endothelial cells is very important for the study of retinal angiogenesis disease.Human retinal microvascular endothelial cells have been successfully cultured in previous studies,but further improvement of the culture method to harvest higher yields and purity cells is still needed.Objective This study was to design a modified method to isolate and purify human retinal microvascular endothelial cells much easily and quickly,and to compare the expression of specific markers of vascular endothelial cells,factor Ⅷ and CD31/CD34 in the cells.Methods The use of human donor eyeballs was approved by the Ethic Commission of Zhongshan Ophthalmic Center of Sun Yat-sen University.The retina tissue from healthy donor was isolated and digested by the two-step digestion method with 2％ trypsin and 0.133％ collagenase Ⅳ.Human retinal microvascular endothelial cells were collected and plated in 60 mm dishes coated by 0.1％ fibronectin and cultured in endothelial cell-specialized medium supplemented with 10％ fetal bovine serum,0.3 mg/L β-endothelial cell growth factor (ECGF) and 100 ng/L sodium heparin.During the culturing,the growth situation of the cells was monitored by morphological observation,and immunohistochemical staining was performed to probe vascular endothelial cell-specific membrane protein CD31,CD34 and factor Ⅷ for identification of the cell purity.Results Human retinal microvascular endothelial cells were isolated successfully from the retina by the twostep digestion method.The primary cultured cells adhered to well 72 hours later and achieved confluence with the typical cobblestone appearance 9 to 10 days after cultured.The cells exhibited the blue nuclei and reddish cytoplasm by regular haematoxylin and eosin stain and showed a strong positive response for CD31,CD34 and factor Ⅷ by immunohistochemistry.The positive dye of CD31 and CD34 was lower than Ⅷ factor in both endothelial cells.Conclusions Modified culture method of human retinal microvascular endothelial cells can improve cell culture result and purify target cells.

Objectives To investigate clinical features and the risk factors for 30-day death in elderly chest pain patients.Methods In the prospective study,514 patients with acute chest pain leading to emergency department visit were selected from March 2012-August 2010 and grouped into elderly group (aged≥65 years,n=309) and non-elderly group (aged< 65 years,n=205).The patient's clinical data during 30-day follow-up period were recorded for analysis and comparison.Multivariate regression analysis was used to investigate the risk factors of death.Results Among 514 cases with acute chest pain,30(5.8%)patients with all-cause death included 24 cases in group of 309 (7.8%) elderly patients and 6 (2.9%) cases in group of 205 non-elderly patients during 30 day follow-up period.Univariate regression analysis showed that female,low SBP,Killips' classification ≥ Ⅱ,high level of serum troponin T and creatinine,coronary artery ischemia were more likely to died during 30 day follow-up period.And female and Killips' classification ≥ Ⅱwere the independent factor for 30-day death in the elderly[OR:3.55 (95%CI:1.00-12.59) and 5.90 (95%CI:1.31-26.63)]respectively.Conclusions Elderly patients with acute chest pain for first emergency department visit are at high risk for 30-day death.Female and cardiac function Killips' classification ≥ Ⅱ,high levels of serum troponin T and creatinine and coronary artery ischemia are associated with 30-day death in patients with acute chest pain for first emergency visit.Female and Killips' classification ≥ Ⅱare the independent risk factor for 30-day death.

Aim To monitor the whole blood trough concentration of cyclosporine A (CsA) in renaltransplant recipients reciving triple therapy with mycophenolate mofetil, cyclosporine andprednisone and to establish an optimal therapeutic window of CsA. Methods Sampleswere measured by specific fluorescence polarization immunoassay. According to the timeafter operation and different therapy plan, the whole blood trough concentration of CsA ineach group was compared with that in control group.Results The optimal therapeuticwindow of CsA with MMF plan was 150~300 ?g? L-1 (less than one month after op-eration), 120~260?g?L-1 (1~

Objective To determine the expression level of deleted in liver cancer-1 (DLC-1)gene in human pituitary tumors, and assess the correlation between DLC-1 expression and clinicopathological parameters in these tumors. Methods DLC-1 and Ki-67 protein expressions were analyzed by immunohistochemistry and DLC-1 mRNA expression by quantitative RT-PCR in 84 pituitary adcnomas and 6 normal human adenohypophyses. Results Immunohistochemical results suggested that DLC-1 expression decreased and Ki-67 expression increased in the pituitary adenomas; Pearson correlation analysis showed that they were negatively correlated (r=-0.764,P=0.001).RT-PCR indicated that DLC-1 mRNA expression decreased in the pituitary adenomas; significantly decreased DLC-1 expression in invasive pituitary adenomas was noted as compared with that in non-invasive ones (P＜0.05). Conclusion Down-regulation of DLC-1 expression may contribute to the tumorigenesis of pituitary adenomas and DLC-1 can be the new target of molecular therapy.

Objective To evaluate whether 18F-fluorothymidine(FLT) can be used to monitor early response to irradiation in colorectal cancer (CRC).Methods SW480 cells were cultured and irradiated with 0, 10, and 20 Gy.Twenty-four hours later, morphological changes, apoptosis, necrosis, proliferation,and cell cycle phases were observed.Uptake of 18F-FLT was measured in these tumors in vitro from 24 h to 72 h after irradiation.The one-way analysis of variance was used to analyze the data.Results Apoptotic and necrotic cells were detected 24 h after radiotherapy.SW480 cells proliferation was significantly delayed after irradiation in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylte-trazolium bromide (MTI) assay.Cell cycle analysis showed that SW480 cells had a decreased fraction of cells in S phase( from 33.23% to 9.24%,then to 5.43% ) and an arrested fraction in G0-G1.After SW480 cells were cultured for60 min, the uptake of 18F-FLT was (5.21 ± 1.60) %; and 24 h after irradiation of 10 Gy, the uptake decreased significantly to (4.27±0.48)% (F=8.253, P=0.009).And 72 h after irradiation, the uptake further decreased significantly to (3.39 ± 0.59) % ( F = 36.715, P＜0.001 ).In tumor tissue, the uptake of 18F-FLT reduced significantly 72 h after radiotherapy (10 Gy:F = 12.388, P = 0.007; 20 Gy:F = 16.744, P = 0.004) and the attenuation degree increased with the radiation dose.Conclusion The uptake of 18F-FLT in SW480 cells or in CRC could reflect the changes of SW480 cells in proliferation, cell cycle re-distribution, cell apoptosis and necrosis.The results suggest that 18F-FLT may be used for monitoring early response to irradiation of CRC.

Objective To evaluate the predictive value of 18F-fluorodeoxyglucose (FDG) and 18F-fluorothymidine(FLT) PET in monitoring the metastatic potential of human colorectal cancer (CRC).Methods Human CRC cell lines SW480 and SW620 were cultured and implanted into nude mice to create CRC models. Tumor growth,metastatic status and survival were assessed in CRC bearing mice. Uptake of 18F-FDG and 18F-FLT in SW480 and SW620 cells was detected In vitro at 0,30,60,90,120 min after incubation. PET images of both tracers were acquired for SW480 and SW620 tumor-bearing mice using the small animal PET at 60 min after tracer injection. Region of interest (ROI) was drawn using Image J software on reconstructed PET images. Immunocytochemistry and Western blot analysis of the tumor tissue were performed. The correlation between tracer uptake and tumor marker expression was evaluated using linear regression. Results Compared with SW620 tumor-bearing mice,SW480 induced tumor grew much faster ( t = - 3.332,P = 0.004),the tumor-bearing mice had more serious dyscrasia ( t = 2.240,P = 0.038 ),shorter survival and higher metastatic rate. In vitro study,the uptake of both 18F-FDG and 18F-FLT in SW620 cells was lower than that in SW480 cells. 18F-FLT uptake was higher than 18F-FDG uptake in both SW480 and SW620 cells. After incubation for 60 min,the uptake of 18F-FDG in SW480 and SW620 cells was ( 1.76 ± 0.87 )% and ( 1.14 ± 0.38 )%,respectively ( t = - 2.507,P = 0.021 ); while the uptake of 18F-FLT in SW480 and SW620 cells was (5.21 ± 1.60)% and (2.90 ± 1.82)%,respectively (t =3.497,P =0.002). In micro-PET study,the 18F-FDG radioactivity ratio of tumor to non-tumor (T/NT) in SW480 and SW620 tumors was 2.69 ± 0.98 and 3.09 ± 1.26 respectively (t =0.657,P =0.524); while T/NT of 18F-FLT in SW480 and SW620 tumors was 3.65 ±0.51 and 2.22 ±0.42 (t =6.491,P ＜0.001 ),respectively. In immunocytochemistry and western blot assay,heat shock protein(HSP) 27,Integrin β3,vascular endothelial growth factor receptor 2 ( VEGFR2 ) and Ki67 were all over expressed in two kinds of tumor cells with different intensities. HSP27 and Integrin β3 expression was higher in SW480 cells than that in SW620 cells. While VEGFR and Ki67 expression was lower in SW480 cells than that in SW620 cells. The uptake of 18F-FLT closely correlated with the expression of HSP27 ( r =0.924,P =0.004) and Integrin β3 (r=0.813,P =0.025). 18F-FDG uptake inversely correlated with the survival of tumor-bearing mice (r =0.500,P=0.017). Conclusions The uptake of 18F-FDGand 18F-FLT may reflect different biological characteritics of CRC. High 18F-FLT uptake in CRC on PET scan may predict high metastatic tendency.

OBJECTIVETo investigate the effects of schisandrin B (Sch-B) on expression of transforming growth factor-beta1 (TGF-beta1) and signal transduction molecule mRNA in rat lungs exposed to SiO2, and explore the intervention mechanism of Sch-B on pulmonary fibrosis induced by SiO2.

METHODSNinety six Wistar rats were randomly divided into control (normal saline) group, SiO2 group and SiO2 plus Sch-B group. The rats were exposed to SiO2 by direct tracheal instillation to establish the silicotic animal models. SiO2 group and SiO2 plus Sch-B group were treated with 1 ml SiO2 (50 mg/ml) for each rat From the first day after model establishment, SiO2 plus Sch-B group were orally given Sch-B (80 mg/kg) a day, control group and silica group were orally given olive oil. On the 3rd, 7th, 14th and 28th days after treatment, 8 rats in each group were sacrificed and samples were collected. The histo-pathological examination of lung was performed by HE staining. The expression levels of TGF-beta1, TGF-betaR II and Smad4 mRNA in the lung tissues were detected by RT-PCR.

RESULTSThe results of histo-pathological examination showed that in SiO2 group, lung tissues were injured obviously; the alveolar inflammation with alveolus interval edema and inflammation cell infiltration appeared on the 3rd and 7th days; the alveolus interval became thicker, became thicker, fibroblast and collagen matrix increased markedly on 14th day; the alveolar structure was damaged, alveolar wall thickened obviously, collagen aggravation and pulmonary fibrosis displayed on 28th day. The alveolar inflammation and pulmonary fibrosis in SiO2 plus Sch-B group were significantly less than those in SiO2 group. The expressions levels of TGF-beta1 TGF-betaR II and Smad4 mRNA (TGF-1beta: 1.03 +/- 0.31, 1.33 +/- 0.39,1.08 +/- 0.26, 0.82 +/- 0.16, TGF-betaR II: 0.65 +/- 0.11, 0.80 +/- 0.16, 0.83 +/- 0.24, 0.62 +/- 0.15, Smad4:0.87 +/- 0.15, 0.68 +/- 0.11, 0.78 +/- 0.19, 0.30 +/- 0.08) in SiO2 group were significantly higher than those in the control group (TGF-beta1:0.59 +/- 0.22, 0.55 +/- 0.25, 0.56 +/- 0.20, 0.55 +/- 0.12, TGR-betaR II :0.28 +/- 0.13, 0.31 +/- 0.15, 0.34 +/- 0.15, 0.27 +/- 0.09, Smad4:0.23 +/- 0.11, 0.40 +/- 0.12, 0.39 +/- 0.12, 0.18 +/- 0.06) (P < 0.01 or P < 0.05), but the expression level of TGF-beta1 mRNA was the highest on the 7th day. The expression levels of TGF-beta1 and Smad4 mRNA (TGF-beta1:0.68 +/- 0.28, 0.88 +/- 0.25, 0.75 +/- 0.11, 0.61 +/- 0.14,Smad4:0.25 +/- 0.12, 0.45 +/- 0.09, 0.44 +/- 0.07, 0.21 +/- 0.04) in SiO2 plus Sch-B group were significantly lower than those in SiO2 group (P < 0.01 or P < 0.05 ), but there were no significant differences of the TGFbetaR II mRNA expression levels between SiO2 group and SiO2 plus Sch-B group.

CONCLUSIONSch-B can reduce the pulmonary fibrosis induced by SiO2 through inhibition of the mRNA express of TGF-beta1 and Smad4 in the lung tissue, modulating the TGF-beta1/Smad4 signal transduction pathway and inhibiting the target gene activation.

Animals ; Cyclooctanes ; pharmacology ; Female ; Lignans ; pharmacology ; Lung ; drug effects ; metabolism ; pathology ; Male ; Polycyclic Compounds ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Signal Transduction ; drug effects ; Silicosis ; metabolism ; pathology ; Smad4 Protein ; genetics ; metabolism ; Transforming Growth Factor beta1 ; genetics ; metabolism