Current therapies for myocardial infarction and congestive heart failure are limited in efficacy or in applicability. The plasticity of adult stem cells and cellular transplantation offer a novel therapeutic approach to improve cardiac function. This review describes the latest progress in research, summarizes recent studies of adult stem cells and their application in myocardial regenerative medicine in China and abroad, and discusses the future directions of cell transplantation as a new therapy to repair injured hearts. (J Geriatr Cardiol 2004;1(2) :77-82. )

AIM: To study the quality standard for Shuwei Capsule (Resina Ferulae, Rhizoma Corydalis, Radix Aucklandiae, etc.). METHODS: Radix Aucklandiae, Rhizoma Cyperi were identified by TLC, and ferulic acid was determined by HPLC. RESULTS: TLC spots developed were fairly clear, and the blank test showed no interference. Ferulic acid showed a good linear relationship in the concentration range of 0.007～0.056?g and the average recovery was up to 98.51%, RSD was 2.64%. CONCLUSION: The method is simple with strong specificity and good reproducibility, and can be used as the quality control of this preparation.

Objective To measure stress distribution of the main bone architecture of the normal adult cadaveric foot and discuss the effect of plantar ligament injury on stress distribution. Methods Seven fresh adult cadaveric feet were used and 10 strain gauges attached to the bones of the longitudinal foot arch,including the calcaneus,navicular,medial cuneiform,1-5 metatarsal trunk,the distal part of the tibia and fibular,respectively.After the loading Was added to 700 N by almighty test machine,resistance strainmeter was used to measure surface strain of these bones.The results were processed statistically. Results The strain was varied based on different bone segments attached and increased with loading.Tensile force was always found at the medial part of the navicular,the distal part of the tibia and fibular,while the others showed compression all the time.Peak strain was found at calcaneus.followed by the second and third metatarsal.Strain on the surface of the bone segments changed greatly with different ligament injury(P＜0.05).Conclusion The bone surface stress of the longitudinal foot arch changes significantly when the plantar ligament is injured.

Objective To establish several human umbilical vein endothelial cell ( HUVEC ) strains with over-ex-pression or low expression of receptor for activated C kinase 1 ( RACK1 ) , which will provide an effective tool for future studying the function of RACK1 in arrhythmia.Methods The full-length cDNA sequence of RACK1 gene was amplified and inserted into pIRES2-EGFP.At the same time, designed and synthesised complementary DNA sequences of 3 pairs of short hairpin structure and a pair of negative control sequence , then subcloned into the plas-mid pGenesil-1 .The HUVEC cells were transfected with these plasmids and screened by using G 418 .And the expression of RACK1 mRNA and protein in the cells were assayed by qRT-PCR and Western blot , respectively . Results RACK1 eukaryotic expression vector and siRNA expression vectors of RACK 1 were constructed success-fully.After a 48 h transfection of HUVEC cells with the recombinant vectors and G 418 selection, the positive cell clones were obtained .qRT-PCR and Western blot showed that over-expression vector and interference vectors could effectively enhanced and knocked-down RACK1 expression in HUVEC strains .Conclusions HUVEC cell strains with over-expression and low expression of RACK 1 have been successfully established .

Objective To investigate the potential of adult mesenchymal stem cells (MSCs) derived from human bone marrow to undergo cardiomyogenic differentiation after exposure to 5-azacytidine (5-aza) in vitro. Methods A small bone marrow aspirate was taken from the iliac crest of human volunteers, and hMSCs were isolated by 1.073g/mL Percoll and propagated in the right cell culturing medium as previously described. The phenotypes of hMSCs were characterized with the use of flow cytometry. The hMSCs were cultured in cell culture medium (as control) and medium mixed with 5-aza for cellular differentiation. We examined by immunohistochemistry at 21 days the inducement of desmin, cardiac-specific cardiac troponin I (cTnI), GATA 4 and connexin-43 respectively. Results The hMSCs are fibroblast-like morphology and express CD44+ CD29+ CD90+ / CD34- CD45- CD31- CD11a. After 5-aza treatment, 20-30% hMSCs connected with adjoining cells and coalesced into myotube structures after 14days. Twenty-one days after 5-aza treatment, immunofluorescence showed that some cells expressed desmin,GATA4, cTnI and connexin-43 in 5,10 μmol/L 5-aza groups, but no cardiac specific protein was found in neither 3μmol/L 5-aza group nor in the control group. The ratio of cTnI positively stained cells in 10 μmol/L group was higher than that in 5 μmol/L group (65.3 ± 4.7% vs 48.2 ± 5.4%, P ＜ 0.05). Electron microscopy revealed that myofilaments were formed. The induced cells expressed cardiac-myosin heavy chain (MyHC) gene by reverse transcription-polymerase chain reaction (RT-PCR). Conclusions Theses findings suggest that hMSCs from adult bone marrow can be differentiated into cardiac-like muscle cells with 5-aza inducement in vitro and the differentiation is in line with the 5-aza concentration. (J Geriatr Cardiol 2004;1(2) :101-107. )

Objective To observe the relationship between apoptosis of K562 cell induced by iron-deprivation and activation of Caspase-3.Methods K562 cells were treated with desferrioxamine(DFO) in different dosages were collected at different time points.K562 cells were labelled with Annexin V/PI,and then the rate of apoptosis was measured by flow cytometry;The activation of Caspase-3 were detected by colorimetric method with pAN labelled substrate;The active protein of Caspase-3 were analyzed by Western blot.Results When K562 cells were treated with different concentrations of DFO,the apoptosis rate and the activity of Caspase-3 increases gradually.When K562 cells were incubated with DFO(50 ?mol/L and 100 ?mol/L) 24 h later,the enzymatic activity of Caspase-3 increases dramatically more than that of control group,and the difference was significantly(P0.05).All those effect above can be counteracted by equal mole concentration of FeCl_3.Conclusion Iron-deprivation maybe induce the apoptosis of K562 cell by chelating intracellular iron and activing Caspase-3.

Traditional Chinese medicine (TCM) dispensing is the final step of TCM used for clinical treatment, the stability of TCM dispensing is the guarantee of good clinical effect. Establishment of effect-constituent equivalence for Chinese herbal pieces based on clinical efficacy, can not only guarantee the stability of TCM dispensing, but also relate to the precision of clinical effect. This study chose Coptidis Rhizoma as the model, established effect-constituent equivalence of Coptidis Rhizoma, based on the effect-constituent index already established by our research group, and taking into consideration of homogeneity of clinical dosage and compliance of decoction, the uniformity of dispensing for different specification of Coptidis Rhizoma decoction pieces was studied. This research model was then applied to guide the specification-optimization of Coptidis Rhizoma and its clinical dispensing. The result indicated, effective constituent equivalence could reflect the fluctuation of specification, dosage and decoction to the fluctuation of efficacy; Optimized Coptidis Rhizoma decoction pieces had the characteristic of high homogeneity as for clinical dispensing, good compliance as for decoction, and high effective constituent equivalence. In conclusion, effective constituent equivalence could improve relevance of methods of TCM dispensing control to clinical effect. Preparated Superior-standard Decoction Pieces based on effective constituent equivalence was featured by good quality and a good practice of adjustable dosage, which could promote the development of TCM decoction pieces toward precision medicine.
Chemistry, Pharmaceutical ; methods ; Coptis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Quality Control ; Rhizome ; chemistry ; Therapeutic Equivalency

Objective To construct and identify a recombinant baculovirus transfer vector of mouse conserved dopamine neurotrophic factor (mCDNF): pFastBacHTb-mCDNF. Methods Mouse total RNA was isolated by using Trizol reagent, and then, first-strand cDNAs were synthesized by reverse transcriptase. Overall length of CDNF (564 bp) was amplified by two rounds of PCR introducing appropriate restriction sites (BamH Ⅰ, Xho Ⅰ). The PCR products were cloned into pGEM-T vector and sequenced to confirm PCR fidelity. The mCDNF was sub-cloned into pFastBacHTb vector to create pFastBacHTb-mCDNF vector, then the vector was transferred into the E. coli DH5α competent cells. The clone was selected using amicillin resistance and then this vector was sequenced and identified by double digests. Results Agarose gel electrophoresis after RT-PCR showed a 564 bp band being consistent with the anticipation size. Positive clone of pGEM-T-CDNF was screened by blue/white and antibiotic resistance selection. Recombinant plasmid pGEM-T-mCDNF was identified by PCR and sequence.Recombinant plsmid pGEM-T-mCDNF and pFastBacHTb vector were cut by BamH Ⅰ and XhoⅠ restriction enzyme, and then, recombinant plasmid pFastBacHTb-mCDNF was constructed and successfully identified by double digestion of Xho Ⅰ and BamH Ⅰ restriction enzyme or single digestion of BamH Ⅰ, PCR and sequence. Conclusion We successfully constructe the recombinant baculovirus transfer vector pFastBacHTb-mCDNF, laying the foundation for further research of this neurotrophic factor.

Objective To investigate the diagnostic accuracies among laparoscopic ultrasonography (LUS),CT,MRCP and transabdominal ultrasonography in secondary choledocholithiasis,and to compare the procedural efficacy of LUS carried out by surgeons assisted by ultrasound physicians,and by surgeons alone.Methods Forty-two patients underwent laparoscopic transcystic common bile duct exploration (LTCBDE) in Beijing Luhe Hospital,Capital Medical University.All these patients underwent LUS examination.In 26 patients,LUS was carried out by surgeons alone while in 16 patients it was assisted by ultrasound physicians.The results of intraoperative choledochoscopy were used to verify the results in the two groups in scan time,and in its accuracies when compared with CT,MRCP and preoperative abdominal ultrasound.Results The accuracy of LUS was 92.9％,which was significantly better than that of CT (73.8％) and transabdominal ultrasonography (23.8％,P ＜0.05).It was also better than that of MRCP (89.7％),though the difference was not significant (P ＞ 0.05).Surgeons alone were faster than ultrasound physicians in performing LUS [(8.5 ± 3.0) min vs (13.2 ± 4.6) min,P ＜ 0.05].There were no statistically differences between the two groups in accuracy (92.3％ vs 93.8％,P ＞ 0.05).Conclusion LUS diagnosed common bile duct stones by surgeons who had adequate ultrasound training,with a high accuracy rate and good efficiency.

China still has more than 30,000 patients of advanced schistosomiasis while new cases being reported consistently. D-dimer is a fibrin degradation product. As ascites being the dominating symptom in advanced schistosomiasis, the present study aimed to explore a prediction model of ascites with D-dimer and other clinical easy-achievable indicators. A case-control study nested in a prospective cohort was conducted in schistosomiasis-endemic area of southern China. A total of 291 patients of advanced schistosomiasis were first investigated in 2013 and further followed in 2014. Information on clinical history, physical examination, and abdominal ultrasonography, including the symptom of ascites was repeatedly collected. Result showed 44 patients having ascites. Most of the patients' ascites were confined in the kidney area with median area of 20 mm². The level of plasma D-dimer and pertinent liver function indicators were measured at the initial investigation in 2013. Compared with those without ascites, cases with ascites had significantly higher levels of D-dimer (0.71±2.44 μg/L vs 0.48±2.12 μg/L, P=0.005), as well ALB (44.5 vs 46.2, g/L) and Type IV collagen (50.04 vs 44.50 μg/L). Receiver operating characteristic curve analyses indicated a moderate predictive value of D-dimer by its own area under curve (AUC) of 0.64 (95% CI: 0.54–0.73) and the cutoff value as 0.81 μg/L. Dichotomized by the cutoff level, D-dimer along with other categorical variables generated a prediction model with AUC of 0.76 (95% CI: 0.68–0.89). Risks of patients with specific characteristics in the prediction model were summarized. Our study suggests that the plasma D-dimer level is a reliable predictor for incident ascites in advanced schistosomiasis japonica patients.
Area Under Curve ; Ascites* ; Case-Control Studies ; China ; Cohort Studies ; Collagen Type IV ; Fibrin ; Humans ; Kidney ; Liver ; Physical Examination ; Plasma* ; Prospective Studies* ; ROC Curve ; Schistosoma japonicum ; Schistosomiasis japonica* ; Schistosomiasis* ; Ultrasonography