Insertion of foreign genes into the genome map of Chinese herb always produces the intended beneficial effects, but also the unintended effects. Analysis of genomic function and metabolic end products based on the principle of substantial can reveal the influence on the quality of herbs, which is able to provide valuable refer-ence for the following evaluation on pharmacology and toxicology.

Objective To investigate the etiological factors, clinical features and prognosis of non- traumatic rhabdomyolysis(RML). Methods The medical records of 13 non-traumatic RML patients hospital- ized between 1995-2006 were reviewed. The etiological, clinical, laboratory and therapeutic data were anal- ysed. Results Among 13 patients with non-traumatic RML, multiple factors were responsible for rhabdomyol- ysis in eight patients and single etiologic factor in 5 patients. Different etiological factors were identified, in- cluding 6 with excessive exercise, 3 with hyperpyrexia, 3 with drugs(including illicit drugs, fenofibrate, cy- closporine), 3 complicated with inflammatory myopathy and 2 with limbs compression. Nine patients had myal- gia and muscle weakness, 6 patients had abnormality in nervous system, 4 patients had hyperpyrexia, 3 pa- tients had digestive symptoms. Nine patients were complicated by coagulation disorders and 6 with acute renal failure(ARF). The serum levels of creatine kinase(CK)were decreased to normal within one month in 6 patients, the patient whose rhabdomyolysis was induced by fenofibrate with diabetes and chronic renal failure showed to inadequate decrease within 60 days. Three patients whose problem was induced by inflammatory myopathy, CK levels decreased within 4 months and 6 months in 2 patients, respectively, but CK level was not returned to normal level in one patient during the 80 follow-up days. Three patients died from multiple causes, such as ARF, coagulation disorders,electrolyte and metabolic disturbances. Conclusion Excessive exercise is the most common cause of non-traumatic RML, followed by drugs and inflammatory myopathy. The prognosis is poor in patients with multiple etiological factors and ARF.

OBJECTIVETo investigate the mechanism of transcription regulation of GLI3 gene in idiopathic congenital talipes equinovarus.

METHODSpGL3-Gli3 luciferase report vectors were constructed, and the activity of Gli3 promoter was explored. A P-Match software was used to analyze the sequence upstream of the transcription start site of rat Gli3 gene, which was subsequently verified with chromatin immunoprecipitation assay (CHIP) and electrophoretic mobility shift assay (EMSA). Expression of the Gli3 gene was analyzed in L6 cells transfected with Hoxd13 small interference RNA(siRNA) and Hoxd13 expression vectors.

RESULTSThe 5' region of rat Gli3 gene contains two potential binding sites for the Hoxd13 protein. CHIP and EMSA assays both confirmed that Hoxd13 can directly bind with site 2. As shown in L6 cells, expression of Gli3 may be enhanced with silencing of Hoxd13, whilst exogenous expression of Hoxd13 can down-regulate transcription of Gli3.

CONCLUSIONHoxd13 can directly regulate the expression of Gli3 gene through a Hoxd13 binding site in the limb of rat embryo.

Animals ; Base Sequence ; Clubfoot ; genetics ; Gene Expression Regulation ; Homeodomain Proteins ; genetics ; Kruppel-Like Transcription Factors ; genetics ; Molecular Sequence Data ; Rats ; Rats, Wistar ; Transcription Factors ; genetics ; Transcription, Genetic ; Zinc Finger Protein Gli3

OBJECTIVETo study the effects of sodium butyrate (SB) on growth, apoptosis and telomerase activity in Hep-2 cells.

METHODSGrowth inhibition effect of SB on Hep-2 cells was assessed by methyl thiazolyl tetrazolium (MTT) assay. Morphological alterations were observed by electronic microscope. Cell apoptosis was confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL) method, DNA fragmentation and flow cytometry (FCM). Cell cycle was analyzed by FCM. Telomerase activity was examined by telomeric repeat amplification protocol (TRAP)-silver staining. The expression status of telomerase subunits was analyzed by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSA time-and dose-dependent inhibition was detected in cells treated with SB. Typical morphological changes of apoptotic cells were observed under electronic microscopy. The characteristic DNA fragmentation of apoptotic cells was detected by agarose gel electrophoresis. Apoptosis and the changes of cell cycle were confirmed by TUNEL method and FCM. The apoptosis indexes of the cells before treatment and at 72 h after SB (2.5 mmol/L) treatment were 2.27 +/- 1.18 and 33.50 +/- 2.75 respectively, the apoptosis rates were 2. 86% and 31. 28% respectively, the proportion of the cells at G0/G1 stage were 50.38% and 70.88% respectively, the proportion of the cells at S stage were 27.40% and 8.20% respectively, and the proliferation indexes of the cells were 49.62% and 29.12% respectively. Telomerase activity and expression level of human telomerase reverse transcriptase (hTERT), the key subunit of telomerase, decreased after SB treatment. No significant changes were observed in the expression of human telomerase RNA (hTR) and human telomerase associated protein (hTP1), the other two subunit of telomerase.

CONCLUSIONSB could inhibit growth of Hep-2 cells and induce apoptosis in the cells, and inhibit telomerase activity by decrease expression level of hTERT.

Apoptosis ; drug effects ; Butyrates ; pharmacology ; Cell Cycle ; drug effects ; Hep G2 Cells ; Humans ; Sodium ; pharmacology ; Telomerase ; metabolism

Objective The aim of this study is to investigate the role of angiotensin-converting enzyme (ACE)gene susceptibility to systemic lupus erythematosus(SLE)by familial studies.Methods PCR-based re- striction fragment length polymorphism(RFLP)was applied to genotype single nucleotide polymorphism(SNP) G261T of the ACE gene.A total of 119 patients with SLE from 119 families were recruited.In addition,316 family members of these patients were also genotyped.A family-based association study was carried out to ex- plore the association between gene polymorphism and SLE.We studied the SNP encoding non-synonymous substitution in the ACE gene with respect to genetic susceptibility to SLE.Results Among 119 SLE patients. the frequency of ACEG261TG,T alleles was 44.8%.55.2% respectively,the frequency of ACEG261T GG,GT and TT genotypes was 13.9%,62.0%,24.1% respectively,Univariate(single-marker)family-based association test(FBAT)demonstrated that variant alleles at the SNP,rs4303,exon 5 of ACE gene were significantly asso- ciated with genetic susceptibility to SLE in Additive Model(Z=2.877,P=0.004),Dominant Model(Z=2.557, P=0.011).Recessive Model(Z=2.202,P=0.028).Transmission-disequilibrium test(TDT)and sib transmission -disequilibrium test(STDT)showed an excess of the allele of T from heterozygous parents to affected offspring or higher frequency of the allele of T in the patients than their normal siblings(X~2=11.66,P=0.001).Conclu- sion Our findings suggest that the ACE gene may he the susceptible gene to SLE in Chinese population,and the individuals carrying ACE-261T allele is significantly associated with susceptibility to SLE.

Objective To explore human umbilical cord blood mesenchymal stem cells(MSCs)colonization in hypoxic-ischemic encephalopathy(HIE)rats brain.Methods Models of 7-day-old newborn rats with HIE brain injury were established.Meanwhile,on the same day,MSCs were transplanted with Hoechst 33258 for 24 hours into rats models marked by flurescent nucleotide dye injected through caudal vein or with stereotactic instrument.After 15-30 days,then MSCs were detected with fluorescene microscope.Results With the improved rice methods,HIE animal model was successfully attained.Majority of MSCs were distributed in the cortex,hippocampus of the lesioned hemisphere,especially in the forehead.And abtained a good fusion with HIE rats brain tissue.Conclusion Human umbilical cord blood MSCs can be cultured,when transplanted into the HIE rats model,they can move into intracranial,and integer with rats brain tissue.

Objective To investigate the possibility of inducing mesenchymal stem cells(MSCs)from human umbilical cord Wharton's Jelly to differentiate into spermatogonia.Methods To isolate,culture and purify MSCs with adherent method,the growth and proliferation of human umbilical cord-derived MSCs were observed,and their immunophenotypes were determined by flow cytometry;MSCs of the third generation were divided into 2 groups to be induced and cultured,MSCs of the control group were cultured in basal medium,while those of the experimental group with conditional medium.The morphologic and ultrastructure changes of control group and experimental group cells were compared with phase contrast microscopy,electron microscopy(EM)and transmission electron microscope(TEM)respectively ;the spermatogonial cells differentiated were then evaluated by immunohistochemistry stained for CD117and CD49f ;the method of Western-blot was used to test if the cells induced could express CD49f.Results A population of MSCs were isolated from human umbilical Wharton's Jelly;they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling;After induction,the shape of MSCs changed greatly from the fibroblast to the round,even familiar to the tadpole;expressed the known molecular markers of spermatogonial cells,such as CD49f,CD117.Conclusion The induced MSCs not only undergo spfermatogonial-cell like morphologic changes,ultramicrostructure mature with increasing cell organs,but also express the spermatogonial cell markers,which show that human umbilical cord derived MSCs are capable of differentiating into spermatogonial cell.

Objective To observe clone ability of human umbilical cord mesenchymal stem cells (MSCs) into infertile mouse seminife-rous tubules and the effects of MSCs on reproductive function.Methods Busulfan was used to destroy endogenous spermatogenesis of the recipient mice.To isolate,culture and purify MSCs with adherent method before marked with Brdu and Hoechst 33258 respectively,and then transplanted into the seminiferous tubules by microinjection.The survival of MSCs in recipient testes were evaluated by immunohistochemistry stained for Brdu and Fluorescent microscopy for Hoechst 33258 observation at different times.The diameter of seminiferous tubules was detected with HMIAS-2000 high-definition colored analyzing system for medical pictures.SPSS 13.0 software was used to analyze the data.Results The dosage of Busulfan resulted in 15% death in the mice,the testis of survived mice showed only basilar membrane in seminiferous tubules after 4 weeks.A lot of purified MSCs were obtained at the third generation and transplantation them into mouse seminiferous tubules survive for at least 4 months and appear to migration.The average diameter in experimental groups were higher than those in controls not only on 26 days but also on 120 days(P

OBJECTIVETo observe the effect of gastric dynamics by transcutaneous electrical acupoint stimulation (TEAS) combined general anesthesia when controlled hypotension dropped to 60% of the mean arterial prenssure (MAP) baseline, and to provide experimental evidence for organ protection in clinical controlled hypotension.

METHODSEighteen male beagles were randomly divided into three groups, the general anesthesia group (blank), the general anesthesia induced controlled hypotension group (control), and the general anesthesia combined TEAS induced controlled hypotension group (experiment), 6 in each group. Controlled hypotension was performed in the latter two groups with isoflurane inhalation and intravenous injection of sodium nitroprusside (SNP). The mean arterial pressure (MAP) was lowered to 60% of the MAP baseline and kept for 60 min. Controlled hypotension was not performed in Beagles of the control group. For Beagles in the experiment group, TEAS [2/100 Hz, (4 ± 1) mA] was applied to bilateral Hegu (LI4), Quchi (LI11), Zusanli (ST36), and Sanyinjiao (SP6) from stable physiological conditions to the end of maintaining stages. Changes of EGG frequencies and EGG amplitudes were monitored. Serum levels of gastrin (GAS) and motilin (MTL) were also detected at corresponding time points during and after experiment.

RESULTSAs for the pressure control effect of TEAS combined general anesthesia in the controlled hypotension, during the process of controlled hypotension (T1-T4), MAP levels of two controlled pressure groups remained relatively stable, and were kept at 60% of the MAP baseline. When the blood pressure dropped to the target low MAP and maintained at 60 min (T1-T4), EGG amplitudes of Beagles in all the three groups showed decreasing tendency. But it was more obviously lower than its basic level in the control group (P <0.05), while it was not obviously decreased in the experiment group (P < 0.05). EGG frequencies of Beagles in all the three groups showed no obvious change during this stage. By the end of the MAP rising stage (T8), the EGG amplitude of the experimental group was significantly higher than that of the control group and the blank group (P < 0.05), while it didn' t show any obvious increase in the control group. During this period, EGG frequencies of the two controlled hypotension groups decreased more than those of the blank group. Two h after rising blood pressure (at T9), EGG amplitudes and frequencies in the two controlled hypotension groups basically restored to their respective baselines and levels of the blank group at T9. At 2 h (T9) after controlled hypotension, serum levels of GAS and MTL were lower than those of basic levels in the two controlled hypotension groups (P <0.05). However, serum levels of GAS and MTL had an increasing trend in the two controlled hypotension groups at 24-72 h (T10-T12). Besides, the increasing speed and amplitude was better in experiment group than in the control group at T10-T12. However, there was no statistical difference between the two groups (P > 0.05). At 72 h (T12) serum levels of GAS and MTL had basically restored to their basic levels in the two controlled hypotension groups and that of the blank control group.

CONCLUSIONEGG amplitudes could be effectively improved in TEAS combined general anesthesia for controlled hypotension at 60% of the MAP baseline, the recovery of the serum GAS level accelerated, gastric power improved and stomach protected.

Acupuncture Points ; Anesthesia ; methods ; Anesthesia, General ; Animals ; Arteries ; Dogs ; Gastrins ; Hypotension, Controlled ; Male ; Motilin ; Nitroprusside ; Transcutaneous Electric Nerve Stimulation

Objective To describe the prevalence,clinical subtypes and associated risk factors of stroke in urban and rural workers from Lhasa,Tibet,and provide the evidence of prevention and treatment.Methods A retrospective,population-based descriptive analysis of 81298 urban and rural workers in the Cities Staff Medical Insurance Registry was performed; stroke and myocardial infarction were defined by the International Classification of Disease(ICD)-10 coded from hospital records; the prevalence of these 2 dieases were compared,and clinical subtypes and associated risk factors of stroke in these stroke patients were analyzed.Results Stroke patients(n=165)were included in the study with a prevalence of 202.96/100,000(95％CI:170.14-231.32),and this number was significantly larger than that of patients with myocardial infarction(n=14),enjoying a prevalence of 17/100,000.The most common subtype was cerebral infarction,followed by cerebral hemorrhage and subaraclnoid hemorrhage; the prevalence of stroke was significantly different in patients with different ages(ranged from 30 to 89,P＜0.05),and the older of the population,the higher prevalence of the stroke; male population had an obviously higher prevalence of stroke than female population,and the population from Han nationality had an obviously higher prevalence of stroke than population from Tibetan nationality (P＜0.05).Conclusion Stroke is the primary fatal disease for the elderly population from Lhasa,and the primary subtype of stroke is cerebral ischemia; its associated risk factors include age,gender and nationality; and combined hypertension is the main risk factor.