Chiropractic plus plum-blossom needling combined with flexibility training on attention deficit in mentally-retarded adolescents / 中国针灸

OBJECTIVE@#To observe the clinical efficacy of chiropractic plus plum-blossom needling combined with flexibility training for attention deficit in mentally-retarded adolescents.@*METHODS@#Thirty adolescents with mild mental retardation were randomly divided into a medical rehabilitation plus flexibility training group (10 cases, 2 cases dropped off), a flexibility training group (10 cases, 1 case dropped off) and a control group (10 cases). The patients in the flexibility training group received flexibility training, once every other day, 3 times a week for 12 weeks. The patients in the medical rehabilitation plus flexibility training group received chiropractic and plum-blossom needling at Baihui (GV 20) and Sishencong (EX-HN 1) on the basis of the treatment in the flexibility training group, once every other day, 3 times a week for 12 weeks. The patients in the control group did not receive any targeted physical training and medical rehabilitation. Tobii Pro Spectrum eye movement instrument was used to test the attention concentration (T), attention span (M), attention transfer (γ%) and attention distribution (η).@*RESULTS@#Compared before treatment, T and M in the medical rehabilitation plus flexibility training group and the flexibility training group were increased after treatment (P<0.01, P<0.05), and γ% in the medical rehabilitation plus flexibility training group was increased after treatment (P<0.05). The increasing range of T, M and γ% in the medical rehabilitation plus flexibility training group and the flexibility training group was greater than that in the control group (P<0.01), and the increasing range of T and γ% in the medical rehabilitation plus flexibility training group was greater than that in the flexibility training group (P<0.05).@*CONCLUSION@#The chiropractic plus plum blossom needling combined with flexibility training can improve the attention deficit in mentally-retarded adolescents.

Advances of targeted delivery vectors for inducing browning of white adipose tissue / 药学学报

Obesity has become an important inducer of many public diseases such as diabetes, endocrine disorders, and so on. Anti-obesity treatment has become a hot topic. Inhibiting fat synthesis and promoting fat decomposition are important ways of drug anti-obesity treatment. With the in-depth study of the distribution, morphology and function of adipose tissue, brown adipose tissue containing multi-compartment fat drops and rich mitochondria have attracted people's attention. Beige adipocytes which are similar to brown adipocytes in morphology and function have aroused great interest, such cells can be transformed from white adipocytes by external stimulation or browning agents. This process is called "white fat browning". The expression of promoting energy consumption proteins in these cells increase, so that the function of adipocytes changes from energy storage to energy consumption to increase excessive energy consumption in the body and reduce lipid accumulation. The browning of white adipose tissue has brought new ideas for obesity treatment, but the systemic administration of browning agent has the risk of adverse reactions to non-target tissues such as heart and central nervous system, which limits its application in inducing white fat browning. Browning agents to white adipose tissue can reduce its adverse reactions and improve its bioavailability by constructing a drug delivery system targeting white adipose tissue. In this review, the mechanism on browning of white adipose tissue, the commonly used browning agents and the targeted delivery carriers that induce browning of white adipose tissue are summarized.

Effect of PEG400 Concentration in Receiving Solution on Drug Transdermal Test in Vitro / 中国实验方剂学杂志

Objective: To investigate the effect of varying concentrations of polyethylene glycol(PEG)400 in receiving solution on in vitro transdermal test of drugs. Method: 5-Fluorouracil(5-FU) was selected as a model drug,by preparing different concentrations of PEG400-phosphate buffer solution(PBS) as the receiving solution,the receiving chamber did not add drug,the excised rat skins were treated with various additives for 12 h,then replaced by PBS and added the saturated model drug into the donor compartment to determine the transdermal parameters of the drug.Meanwhile,scanning electron microscopy(SEM) was employed to monitor the effect of PEG400 with different concentration on the stratum corneum of rat skin. Result: The 10%,15% and 40% PEG400-PBS groups had no significant effect on in vitro transdermal absorption parameters of the 5-FU.The steady transdermal rate and cumulative penetration rate of the drug in 20% and 30% PEG400-PBS groups were significantly higher than that in the PBS group(PPConclusion: In the rat skin transdermal test,the concentration of PEG400 in receiving solution should be controlled below 20%.

Preparation and quality evaluation of levocarnitine thermosensitive in situ gel / 药学学报

In this paper, a new type of preparation for treatment of initial dry eye disease, thermosensitive in situ gel, was prepared using levocarnitine as a model drug. Poloxamer 407 and poloxamer 188 were used as the gel matrix, and sodium hyaluronate and sodium carboxymethylcellulose were used as bioadhesive materials. Gelation temperature was determined by a rotor method and the prescription was optimized by central composite design-response surface methodology. The pH value, viscosity value and gelation temperature of the optimal prescription were measured. The release of the drug in vitro was examined by dialysis membrane permeation, and retention time of the thermosensitive in situ gel preparation on the rabbit's ocular surface was observed by a slit lamp microscope. The results showed that the dosage of the poloxamer 407 and poloxamer 188 were 20.81% and 3.46%, respectively, and sodium hyaluronate was 0.02%, sodium carboxymethyl cellulose was 0.10% of the optimal formulation of levocarnitine thermosensitive in situ gel. The pH value was 6.90 ± 0.06 at room temperature and the viscosity value started to rise sharply at 27 ℃ of the optimal formulation. The gelation temperature of the optimal preparation before and after dilution by simulated tear fluid were (26.37 ± 0.06) ℃ and (33.57 ± 0.21) ℃, respectively. In the first 240 min, in vitro release rate per unit area of levocarnitine thermosensitive in situ gel was lower than that of solution (P<0.05), and after 600 min, the cumulative release rate of levocarnitine thermosensitive in situ gel could reach more than 80%. The retention time of the thermosensitive in situ gel preparation on rabbit's ocular surface reached about 25 min, at least 5 times as much as that of the solution. The animal experiment was conducted following the National Institutes of Health Guidelines for the use of experimental animals, and approved by the Ethics Committee of the Experimental Animal Center of Beijing University of Chinese Medicine. The levocarnitine thermosensitive in situ gel showed good characteristics and sustained release property and significantly improved the retention time of the drug on the rabbit's ocular surface.

Protective effect of early intervention with lipoxin A4 on septic mice / 中国当代儿科杂志

OBJECTIVE@#To study the effect of early intervention with lipoxin A4 (LXA4) on septic mice.@*METHODS@#Healthy male Balb/c mice aged 6-8 weeks were randomly divided into sham-operation group, sepsis group, 1-hour intervention group (intervention at 1 hour after sepsis), and 6-hour intervention group (intervention at 6 hours after sepsis) (n=8 each). A sepsis model was prepared by cecal ligation and puncture. The intervention groups received LXA4 at 0.01 μg/g body weight 1 or 6 hours after the model was established. Blood was taken from eyeballs at 24 hours after operation. Peritoneal lavage fluid and liver and lung tissue samples were collected. The bacterial colonies of whole blood and peritoneal lavage fluid were counted by dilution plating. The serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1) were determined by cytometric bead array. The serum level of high mobility group box-1 (HGMB1) was determined using ELISA. The percentages of macrophages and neutrophils in peritoneal lavage fluid were determined by flow cytometry. Paraffin sectioning and hematoxylin-eosin staining were performed for the liver and lung tissue samples to observe pathological damage.@*RESULTS@#Compared with the sham-operation group, the sepsis group had a significantly decreased percentage of macrophages and a significantly increased percentage of neutrophils in peritoneal lavage fluid (P0.05). Compared with the 6-hour intervention group, the 1-hour intervention group had a significantly decreased serum level of HMGB1 (P0.05).@*CONCLUSIONS@#Early intervention with LXA4 may attenuate liver and lung injuries in septic mice, which may be explained by the decrease in serum levels of IL-6, TNF-α, MCP-1, and HMGB1, and it also may reduce the bacterial dissemination in the whole blood of septic mice, which may be explained by the increase in the percentage of peritoneal macrophages.

Effect of lipoxin A4 on the expression of Toll-like receptor 4 and TNF receptor-associated factor 6 in the liver of obese rats with sepsis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the protective effect of lipoxin A4 (LXA4) against sepsis induced by lipopolysaccharide (LPS) in rats with obesity and its effect on the expression of Toll-like receptor 4 (TLR4) and TNF receptor-associated factor 6 (TRAF6) in the liver.</p><p><b>METHODS</b>A total of 60 male Sprague-Dawley rats aged three weeks were randomly divided into a normal group and an obesity group, with 30 rats in each group. A rat model of obesity was established by high-fat diet. Each of the two groups was further randomly divided into control group, sepsis group, and LXA4 group, and 8 rats were selected from each group. The rats in the control, sepsis, and LXA4 groups were treated with intraperitoneal injection of normal saline, LPS, and LXA4+LPS respectively. Twelve hours later, blood samples were collected from the heart and liver tissue samples were also collected. ELISA was used to measure the serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Western blot was used to measure the protein expression of TLR4 and TRAF6 in liver tissue. Quantitative real-time PCR was used to measure the mRNA expression of TLR4 and TRAF6.</p><p><b>RESULTS</b>After being fed with high-fat diet for 6 weeks, the obesity group had significantly higher average weight and Lee's index than the normal group (P<0.05). Compared with the normal group, the obesity group had significant increases in the serum levels of IL-6 and TNF-α (P<0.05). In the normal group or the obesity group, the sepsis subgroup had significant increases in the serum levels of IL-6 and TNF-α compared with the control subgroup (P<0.05), while the LXA4 subgroup had significant reductions in the two indices compared with the sepsis subgroup (P<0.05). Compared with the normal group, the obesity group had significant increases in the protein and mRNA expression of TLR4 and TRAF6 (P<0.05). In the normal group or the obesity group, the sepsis subgroup had significant increases in the protein and mRNA expression of TLR4 and TRAF6 compared with the control subgroup (P<0.05). Compared with the sepsis subgroup, the LXA4 subgroup had significant reductions in the protein and mRNA expression of TLR4 and TRAF6 (P<0.05).</p><p><b>CONCLUSIONS</b>LXA4 can reduce the serum levels of IL-6 and TNF-α and alleviate inflammatory response. LXA4 can inhibit the expression of TLR4 and TRAF6 in the liver of septic rats, possibly by inhibiting the TLR4 signaling pathway.</p>

Clinical Value of Presepsin in the Diagnosis and Prognosis of Bacterial Infection in Cirrhosis / 现代检验医学杂志

Objective To observe and evaluate the clinical value of presepsin level in cirrhosis-associated bacterial infections. Methods 30 cirrhosis patients with bacterial infections were enrolled as study group.At admission,60 cirrhosis patients without bacterial infections were enrolled as control group.The difference of blood presepsin level between two groups were analyszed and compared the relationship between level of presepsin patients.The level of presepsin of survival and death group were compared and analyszed risk factors for the prognosis of bacterial infection by COX multi-factor analysis.The sensitivity and specificity were compared by PCT,CRP and presepsin detection.Results The blood presepsin level of study group was 1 002.3(575.1 2~149.5)pg/ml,the blood presepsin level of control group was 475.0(332.7~680.2)pg/ml. The presepsin of study group was significantly higher than control group,the difference was statistically significant(t=1.865,P<0.05).The presepsin level of death patients was significantly higher than survival patients(t=5.875,P<0.05). COX multi-factor analysis showed that presepsin levels were independent risk factors affecting the prognosis of bacterial in-fection in patients with liver cirrhosis.There were statistically significant of sensitivity and specific degrees between presep-sin and CRP(P<0.05).Conclusion Presepsin level is a valuable new biomarker for defining severe infections in cirrhosis. The increase of Presepsin level can be used as a judgment is an important index indicator of liver cirrhosis bacterial infection. It was worthy of clinical attention.

Expression of DOG-1 and C-erbB-2 in papillary thyroid carcinoma and their clinical significance / 临床与实验病理学杂志

Purpose To investigate the expression and clinical significance of DOG-1 and C-erbB-2 in papillary thyroid carcinoma.Methods Immunohistochemical SP method was used to detect the expression of DOG-1 and C-erbB-2 protein in 48 cases of papillary thyroid carcinoma and 30 cases of benign thyroid lesions (15 cases of nodular goiter and 15 cases of Hashimoto's thyroiditis).Results The DOG-1 positive rate of thyroid papillary carcinoma (27.08％,13/48) was significantly higher than that of benign lesions (0,0/30),the difference was significant (P ＜ 0.05).The C-erbB-2 positive rate (39.58％,19/48) was significantly higher than that of benign lesions (3.33％,1/30),the difference was significant (P ＜ 0.05).Positive expression of DOG-1 as well as C-erbB-2 correlated with advanced TNM and presence of lymph node metastasis.The expression of DOG-1 and C-erbB-2 in patients with multifocal carcinomas combined with lymphatic metastasis showed a significant positive correlation(r =0.503,P =0.024).Conclusion The data suggest that DOG-1 and C-erbB-2 contribute to the pathogenesis and progress of thyroid papillary carcinoma.Our results introduce DOG-1 combined with C-erbB-2 as a promising biomarker for recurrence prediction and target therapy.

Expression, subcellular localization and characterization of dammarenediol-Ⅱ synthase of Panax ginseng in Saccharomyces cerevisiae / 药学学报

To study the expression and subcellular localization of recombinant dammarenediol-Ⅱ synthase (DS) in Saccharomyces cerevisiae, the dammarenediol-Ⅱ synthase gene ds was cloned from Panax ginseng, and the gene ds was fused with the gene of green fluorescent protein to obtain the fusion gene ds-gfp. The recombinant expression plasmids pESC-HIS-DS and pESC-HIS-DS-GFP were constructed and transformed into S. cerevisiae INVSc1 to obtain recombinant strains INVSc1-DS and INVSc1-DS-GFP. Microsomes of recombinant strains were prepared by differential centrifugation and observed by fluorescence microscope. The green fluorescence was only detected in INVSc1-DS-GFP microsomes, which indicated that DS was a membrane protein. It was also proved that dammarenediol-Ⅱ was produced from the cyclization of 2, 3-oxidosqualene catalyzed by DS through in vitro enzymatic reaction. In addition, our results revealed that the fusion expression of ds with gfp significantly improved the production of dammarenediol-Ⅱ from 7.53 mg·g-1 to 12.24 mg·g-1. This study provides a new strategy in the optimization of the pathway of ginsenosides biosynthesis in S. cerevisiae.

Downregulation of lanosterol synthase gene expression by antisense RNA technology in Saccharomyces cerevisiae / 药学学报

The cyclization of 2,3-oxidosqualene is the key branch point of ergosterol and triterpenoid biosynthesis. Downregulation of 2,3-oxidosqualene metabolic flux to ergosterol in Saccharomyces cerevisiae may redirect the metabolic flux toward the triterpenoid synthetic pathway. In our study, primers were designed according to erg7 gene sequence of S. cerevisiae. Three fragments including 5' long fragment, 5' short fragment and erg7 coding region fragment were amplified by PCR. 5' long fragment consists of the promoter and a part of erg7 coding region sequence. 5' short fragment consists of a part of promoter and a part of erg7 coding region sequence. These fragments were inserted reversely into pESC-URA to construct antisense expression plasmids. The recombinant plasmids were transformed into S. cerevisiae INVSc1 and recombinant strains were screened on the nutritional deficient medium SD-URA. The erg7 expression level of recombinant strains, which harbored antisense expression plasmid of erg7 coding region, was similar to that of INVScl by semi-quantitative PCR detection. But erg7 expression level of recombinant strains, which harbored 5' long antisense fragment and 5' short antisense fragment, was significantly lower than that of the control. The results of TLC and HPLC showed that the ergosterol content of recombinant strains, which harbored 5' long antisense fragment, decreased obviously. The ergosterol contents of the others were almost equal to that of INVSc1. Lanosterol synthase gene expression was downregulated by antisense RNA technology in S. cerevisiae, which lays a foundation for reconstructing triterpenoid metabolic pathway in S. cerevisiae by synthetic biology technology.

Construction of Saccharomyces cerevisiae haploid mutant deficient in lanosterol synthase gene / 药学学报

Lanosterol synthase is encoded by the erg7 gene and catalyzes the cyclization of 2, 3-oxidosqualene, which is a rate-limiting step of the inherent mevalonate (MVA)/ergosterol metabolic pathway in Saccharomyces cerevisiae. The intermediate 2, 3-oxidosqualene is also the precursor of triterpenoids. Therefore, the cyclization of 2, 3-oxidosqualene is the key branch point of ergosterol and triterpenoids biosynthesis. Down-regulation of 2, 3-oxidosqualene metabolic flux to ergosterol in S. cerevisiae may redirect the metabolic flux toward the triterpenoid synthetic pathway reconstructed by the synthetic biology approach. To construct erg7 knockout cassette harboring the loxP-Marker-loxP element, long primers were designed, which were homologous to the sequences of both erg7 ORF and plasmid pUG66. The cassette was transformed into diploid wild strain INVSc1 by LiAc/SS Carrier DNA/PEG method and then erg7 gene haploid deficient mutant was obtained by homologous recombination. The results of semiquantitative PCR and real-time quantitative PCR revealed that erg7 expression level in erg7 gene haploid deficient mutant is one time lower than that in wild strain. The results of TLC and HPLC showed that the ergosterol content in deficient mutant decreased to 42% of that in wild strain.

Research advances of the influence factors of high level expression of recombinant protein in Pichia pastoris / 药学学报

Pichia pastoris is one of the most important systems used in the field of molecular biology for the expression of recombinant proteins. The system has advantages of high expression, high stability, high secretion, easy high-density fermentation and low cost. Many factors affect the expression of recombinant protein, such as gene copy number, codon usage preference, type of promoter, molecular chaperones, glycosylation, signal peptide and fermentation process. In this review, research advances of the above aspects are summarized, which lay a foundation for improving the expression of recombinant proteins in P. pastoris.

Advances in the biosynthesis research of ginsenosides / 药学学报

Ginsenosides are the main active components of medicinal herbs including Panax ginseng and Panax quinquefolium, which have potent effects of anti-tumor, anti-inflammatory, antioxidant and apoptosis inhibition. But the low content of ginsenosides limits its development and usage. At present, how to improve the production of ginsenosides by biological technology has been a new research focus. Some advances in the biosynthesis of ginsenosides by tissue culture and biotransformation have been made in recent years. So far at least twenty genes related to the biosynthesis of ginsenosides from Panax genus plants have been cloned and functionally identified, which has laid a good foundation for the study on the synthetic biology of ginsenosides. This review outlines recent advances in several aspects and is expected to provide a theoretical support to the thorough research of the pathway and regulation of ginsenosides biosynthesis.

Codon optimization and eukaryotic expression analysis of the analgesic peptide gene BmK AngM1 from Buthus martensii Karsch / 药学学报

Codon bias is an important factor which influences heterologous gene expression. Optimizing codon sequence could improve expression level of heterologous gene. In order to improve the expression level of BmK AngM1 gene encoding the analgesic peptide from Buthus martensii Karsch in Pichia pastoris, the codon-optimized BmK AngM1 gene according to its cDNA sequence and the preference codon usage of P. pastoris were cloned into expression vector pPIC9K and then transformed into P. pastoris. The expersion of recombinant BmK AngM1 (rBmK AngM1) was inducced by methanol in the medium, and the expression level of the optimized BmK AngM1 gene was 3.7 times of the native one. These results suggested that the expression of BmK AngM1 in P. pastoris could be successfully improved by codon optimization.

Bone mesenchymal stem cell transplantation repairs glomerular podocytes in rats with puromycin aminonucleoside-induced nephrosis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the effect of bone mesenchymal stem cell (BMSC) transplantation on repair of glomerular podocytes and on the Nephrin expression in rats with puromycin aminonucleoside (PAN) -induced nephrosis.</p><p><b>METHODS</b>Forty-five Sprague-Dawley rats were randomly divided into three groups (n=15 each): a nephrosis model group that received a single intraperitoneal injection of PAN (0.15 mg/g); a BMSC transplantation group that received a single intraperitoneal injection of PAN (0.15 mg/g) followed by BMSC transfusion; a control group that received a single intraperitoneal injection of normal saline. Ten days after injection, the rats were sacrificed. The 24 hrs urinary protein content and serum albumin and cholesterol levels were measured 24 hrs before sacrifice. Changes of glomerular podocytes were observed under an electron microscope. Brdu labeled positive cells in kidneys were measured by immunohistochemical technology. RT-PCR and Western blot were used to assess the expression of mRNA and protein of Nephrin.</p><p><b>RESULTS</b>In the nephrosis model group, urinary protein and blood cholesterol contents increased, plasma albumin content decreased compared with those in the control group. Extensive fusion of podocyte foot processes was observed in the nephrosis model group. The BMSC transplantation group had decreased urinary protein and blood cholesterol contents and increased plasma albumin content compared with the nephrosis model group. Fusion of podocyte foot processes was also improved. Brdu labeled positive cells were seen in kidneys in the BMSC transplantation group, but not in the nephrosis model and the control groups. Nephrin mRNA and protein expression decreased significantly in the nephrosis model group compared with that in the control group. The BMSC transplantation group had increased Nephrin mRNA and protein expression compared with the nephrosis model group.</p><p><b>CONCLUSIONS</b>BMSCs can repair glomerular podocytes in PAN-induced nephrosis rats, and the changes of Nephrin expression may be involved in the process.</p>

Roller compaction research of Banlangen effervsce tablets / 中国中药杂志

<p><b>OBJECTIVE</b>To study the conditions and parameters of roller compaction of Banlangen effervsce tablet.</p><p><b>METHOD</b>The experimentation adopts L9 (3(4)) orthogonal experiment to study the conditions and parameters of roller compaction of Banlangen effervsce tablet; studied factors that included roller pressure, roller speed and moisture content of power, which influence the result of granule yield and granule friability.</p><p><b>RESULT</b>The optimal technique is: roller pressure at 1.5 MPa; roller speed at 15 Hz; moisture content of power at 1.5%.</p><p><b>CONCLUSION</b>The study of roller compaction technique of Banlangen effervsce tablets provides some technicial consults of its research and production.</p>

Review of effervescent technique in pharmaceutics research / 中国中药杂志

Effervescent technique, which can accelerate drug disintegration and dissolution, is usually applied in quick release preparations. Along with the development of pharmaceutical technique and theory, effervescent technique is used more and more extensively to adjust the behavior of drug release, such as in sustained and controlled release preparations, pulsatile drug delivery systems, and so on. This review demonstrated the new applying of effervescent technique in effervescent tablets, stomach floating forms, osmotic pump tablets and pulsatile drug delivery systems, adding to the critical common technique of effervescent forms in drug research. This will be benefit for the further research and development of effervescent technique.

A study of reliability and validity of appraisal questionnaire (Chinese language version) of human immunodeficiency virus positive persons / 中华预防医学杂志

<p><b>OBJECTIVE</b>To evaluate the Chinese language version of the appraisal questionnaire (AQ) for HIV/AIDS (HIV positive persons).</p><p><b>METHODS</b>The scale was translated and adapted into Chinese and then reversely translated into English. The internal consistency reliability, structural validity, differential validity and predictive validity were evaluated by prevalence study in the city of Guangzhou.</p><p><b>RESULTS</b>Cronbach's alpha coefficient achieved a value ranged from 0.530 to 0.886, with satisfied predicted validity. The regression equation accounted for 24.4% of variance in anxiety, and all factors of cognition accounted for 11.4% of variance, among all factors, only factor one had significant influence on anxiety (t=3.838, P<0.001); the regression equation accounted for 38.6% of variance in depression, and all factors of cognition accounted for 26.0% of variance. Factor I and factor II had significant influence on depression (t=5.707, P<0.001; t = -2.876, P<0.01). The results of differential validity suggested the mean scores of factor III were lower in the group with lower education level and monthly salary. Meanwhile, the persons with monthly salary under 300 yuan RMB had higher mean score of factor I, and the persons with lower academic degree had lower mean score of factor II. The principal-components factor analysis yielded three factors with common factor larger than 1.0, which were threat, challenge and controllability; and the three factors accounted for 62.23% of the total variance.</p><p><b>CONCLUSION</b>The AQ Chinese version attained satisfactory reliability and validity. Even considering some essential explanatory words for the Chinese version, the scale might be attempted to use in the population with HIV in our country.</p>

Construction of a gastric cancer related gene GCRG213-specific siRNA expression vector and its effect on gastric cancer cells in vitro / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the effect of gene GCRG213 siRNA transfection into gastric cancer cell line MKN45 cells.</p><p><b>METHODS</b>Two pairs of DNA sequences containing small hairpin structure to GCRG213 were designed and synthesized. The complement form was obtained by annealing and inserted into RNAi expression vector IMG-800. They are IMG-800-1 and IMG-800-2 correspondingly. The recombinant plasmid IMG-800-1, IMG-800-2 and the vector IMG-800 were separately transfected into MKN45 cells conducted by lipofectamine 2000. After G418 selecting, the cells were transfected steadily. Expression of GCRG213 was detected by semi-quantitative RT-PCR and Western Blot. The growth graph of six steady transfected cell cultures was protracted by cell counting. FACS was used to detect the cell cycle, and Annexin V FITC/PI double labeling were used to detect the effects on cell apoptosis in the above-mentioned cells. The clone formation rate in plate and in nude mice was tested to investigate the tumorigenic characteristics of the six steadily transfected cells in vitro and vivo.</p><p><b>RESULTS</b>Through sequencing, two pairs of DNA sequences containing small hairpin structure to GCRG213 were proved to be successfully cloned into siRNA expression vector IMG-800, correspondingly called IMG-800-1 and IMG-800-2. The recombinant plasmid IMG-800-1, IMG-800-2 and vector IMG-800 were transfected separately into MKN45 cells conducted by lipofectamine 2000. After G418 selecting, the cells were transfected steadily. Transfecting the siRNA vector (IMG-800-1, IMG-800-2 ) into the MKN45 cells significantly decreased the expression of GCRG213, at both mRNA and protein levels. The growth graph showed that the growth of IMG-800-1 and IMG-800-2 transfected cells were slower than that of vector transfected cells. The proportion of cells in G2/M and/or S phase decreased in the cells transfected with IMG-800-1 and IMG-800-2 and cell apoptosis increased. The average clone formation rate in vitro decreased in the cells transfected with IMG-800-1 and IMG-800-2, compared with those transfected with vector. In vivo, the time of tumor formation of IMG-800-1 and IMG - 800-2 transducted cells in nude mice was prolonged and the tumor size was smaller.</p><p><b>CONCLUSION</b>GCRG213 SiRNA transfection may induce inhibition of growth and proliferation of tumor cells, promote cell apoptosis, and inhibit the tumorigenicity in vitro and vivo.</p>

Antiapoptotic Interventions in Intracerebrai Hemorrhage / 国际脑血管病杂志

Tne secondary injury may occur in the perihematomal tissue after intracerebral hemorrhage(ICH).Studies have suggested that apoptosis palys an important role in the secondary injury after intracerebral hemorrhage.1he inhibition of apoptosis may significantly improve the neurological deficit after ICH.