With the development of functional genomics, high throughput analysis of genes’ function has been the mainstream of research, and exogenous gene's over expression via Agrobacterium-mediated transformation is the most commonly used method in gene functional analysis.The versatile plant expression vector cassette named pBHT-5 was constructed by the method of site-specific mutagenesis based on pBI121. First of all, the restriction enzyme SfiI recognition site in trfA gene (X00713) which was relevant to plasmid replication and stability was replaced without changing its amino acid composition. And then the SfiIA,SfiIB sites were added between promoter CaMV35s and terminator NOS. The versatile plant expression vector cassette can be directly used to construct plant expression vector containing the full-length genes cloned by Clontech SMARTTM technology, which will raise the efficiency of vector construction. The result will provide basis of new genes’ high throughput screening and functional analysis, then get the new genes functioning in plant osmotic stress resistance.

Objective To assess the efficacy and safety of various spectrum lasers in the treatment of acne vulgaris. Methods Based on the principles and methods of Cochrane systematic reviews, we searched the Cochrane Library (2009, 6 issues), PubMcd, Embase, China Biomedical Literature Database, China Journal Full Text Database, Chinese Scientific Journals Full Text Database. Retrieval time was up to June, 2009. Randomized controlled trials (RCTs) of lasers for acne vulgaris were included. Results Twelve RCTs totaling 367 patients were included. Because the lack of clinical homogeneity, only descriptive analysis was conducted. Acne lesion counts improved significantly with laser therapy. Adverse effects were limited to transient erythema and edema at treatment sites. Treatment-related pains were well tolerated. Conclusions Current evidence demonstrates that all type lasers in treating acne vulgaris is safe and efficacy. However, higher quality RCT research would be needed to verify the effects and status of lasers on acne vulgaris.

This study is to investigate the effect of ethyl gallate on invasion capabilities and its mechanism of breast cancer MDA-MB-231 cells. Using cell adhesion and transwell assay, separately, the effects of ethyl gallate on the invasion of MDA-MB-231 cells were measured. The Akt-NF-κB signal pathway protein expressions were analyzed with Western blot. Also, the mRNA levels of MMP-9 and MMP-2 were analyzed by RT-PCR. Ethyl gallate inhibited the abilities of motility, adhesion and invasion of breast cancer MDA-MB-231 cells in vitro (P<0.05), inhibited the mRNA levels of MMP-9, MMP-2, phosphorylation of AKt and protein expression of NF-κB. It is concluded that ethyl gallate can inhibit the abilities of invasion of breast cancer in vitro by inhibiting the mRNA levels of MMP-9/MMP-2, phosphorylation of Akt and protein expression of NF-κB.
Breast Neoplasms ; pathology ; Cell Adhesion ; drug effects ; Cell Line, Tumor ; Cell Movement ; drug effects ; Gallic Acid ; analogs & derivatives ; pharmacology ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; NF-kappa B ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; Signal Transduction

OBJECTIVETo observe the characteristics of sperm single-stranded DNA breaks (SSB) and double-stranded DNA breaks (DSB) in infertile men, explore the association of DSB with male infertility, and provide a new observation index and idea for the diagnosis and treatment of the disease.

METHODSThis study involved 60 infertile men (infertility group) and 30 normal healthy males with infertile wives (control group). We comparatively analyzed the seminal parameters of the two groups, determined sperm concentration and viability using the computer aided sperm analysis system, measured the sperm survival rate by hypoosmotic swelling (HOS) test, examined sperm morphology by Diff-Quick staining, and detected sperm DNA damage by two-tail comet assay.

RESULTSNine two-tail comet models were established for detecting sperm DNA integrity. Comparisons between the fertility and control groups showed that the sperm DNA fragmentation index (DFI) was (33.8 ± 13.1) vs (16.3 ± 7.9)% (P < 0.01), the SSB-DFI was (19.2 ± 11.4) vs (14.9 ± 7.6)% (P > 0.05), the SSB-DFI/DFI was (56.8 ± 32.4) vs (91.4 ± 27.8)% (P < 0.01), the DSB-DFI was (23.9 +13.4) vs (6.1 ± 2.7)% (P < 0.01), and the DSB-DFI/DFI was (70.8 ± 19.5) vs (37.4 ± 11.3)% (P < 0.01). The optimal cut-off value of DSB-DFI/DFI in the diagnosis of male infertility was 39.5%, with the AUG, sensitivity, and specificity of 0.969, 98.3%, and 90%; that of DSB-DFI was 15.85%, with the AUC, sensitivity, and specificity of 0.912, 86.7%, and 80%; and that of DFI was 18.65%; with the AUC, sensitivity, and specificity of 0.861, 90%, 70%, respectively. In the infertile men, neither SSB-DFI nor SSB-DFI/DFI exhibited any correlation with semen parameters (P > 0.05); DFI was correlated negatively with the percentage of progressively motile sperm, sperm survival rate, and the percentage of morphologically normal sperm (P < 0.05 or P < 0.01), but not correlated with sperm concentration (P > 0.05); both DSB-DFI and DSB-DFI/DFI showed a negative correlation with sperm concentration, sperm survival rate, and the percentages of progressively motile sperm and morphologically normal sperm (P < 0.05 or P < 0.01).

CONCLUSIONDouble-stranded, rather than single-stranded DNA breaks, may be a factor inducing male infertility. The type of sperm DNA strand damage is of much reference value for the assessment of male fertility.

Case-Control Studies ; Comet Assay ; DNA Breaks, Double-Stranded ; DNA Breaks, Single-Stranded ; DNA Fragmentation ; Fertility ; Humans ; Infertility, Male ; diagnosis ; genetics ; Male ; Semen Analysis ; Sensitivity and Specificity ; Sperm Count ; Spermatozoa ; Staining and Labeling

Objective To observe the effect of using Ambroxol hydrochloride(AM)to wash respiratory way to treat neonatal respiratory distress syndrome(NRDS) in ventilator,to explore dynamic changes of respiratory mechanics after using AM to wash respiratory way.Methods Thirty premature infants were chosen according with diagnosis criterion,which were randomly divided into 2 groups: NS group(n=15);AM group(n=15).Both NS and AM groups were treated with Babylog 8 000 ventilator,and routine treating and nursing,NS group was given for washing respiratory way in NS group,whereas AM was done in AM group.Pulmonary compliance(C),time constant(Tc),respiratory resis-tance(R),C20/C and minute volume(MV)were observed in both groups.Blood gas was routinely checked after 1 h ventilation treatment,and X ray was shot after 24 h.Results Pulmonary C significantly increased in weaning than that of beginning ventilation(P0.05),MV significantly increased in group AM than NS,respectively[(0.56?0.12) L/min and(0.35?0.11) L/min(P0.05).But ventilator-treating-time was markedly shorter in group AM than NS,respectively(60.52?6.23) h and(98.21?5.82) h(P

Biomarkers ; blood ; Child ; Chorionic Gonadotropin ; blood ; Follicle Stimulating Hormone ; blood ; Growth Disorders ; etiology ; Humans ; Klinefelter Syndrome ; complications ; diagnosis ; genetics ; Magnetic Resonance Imaging ; Male ; Mediastinal Neoplasms ; complications ; diagnosis ; surgery ; Puberty, Precocious ; diagnosis ; etiology ; Teratoma ; complications ; diagnosis ; surgery ; Testis ; pathology

OBJECTIVETo analyze genetic polymorphisms of Penta D and E loci in Korean racial Chinese of Yanbian area.

METHODSOne hundred unrelated individuals of Korean racial Chinese were analyzed by methods of PCR and restriction fragment length polymorphism.

RESULTSEight alleles and 22 genotypes were observed from Penata D locus, and 16 alleles and 35 genotypes were observed from Penta E locus.

CONCLUSIONThe distribution of two locus genotypes in Korean racial Chinese of Yanbian area met Hardy-Weinberg equilibrium. And the obtained data can be used to individual identity, paternity testing and the study of Korean ethnic group of Chinese population.

Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Ethnic Groups ; genetics ; Humans ; Polymorphism, Genetic

Objective To investigate the association between C2549A polymorphism of leptin gene and the Trp64Arg polymorphism of the β3-adrenergic receptor gene(β3-AR gene) and obesity in Kazak school-age children.Methods Totally 92 obese children and 71 healthy controls were selected from 6 to 12 years old in Kazak school-age children from the area around Urumqi.Genotype of the C2549A polymorphism of leptin gene and the Trp64Arg polymor phism of β3-AR gene were determined by polymerase chain reaction and restriction fragment length polymorphism methods.Results 1.The difference in distribution of the β3-AR gene Trp64Arg genotype of obesity and healthy controls of Kazak children was statistically significant (x2 =10.472,P ＜ 0.05),but the difference in distribution of the alleles was not statistically significant (x2 =3.541,P ＞ 0.05).2.The differences in distribution of the leptin C2549A genotype and alleles of the 2 groups were all statistically significant,and the odds ratio of the alleles(0.608) and 95％ CI 0.380-0.972 suggested that the mutation occurrence of obesity might have certain protective effect.3.The difference in distribution of the genotype of β3-AR gene Trp64Arg polymorphism and leptin gene promoter area C2549A polymorphism in the same individual joint action of 2 groups was statistically significant (x2 =15.978,P ＜ 0.05),but the difference in distribution of the alleles in the same individual joint action of 2 groups was not statistically significant (x2 =6.362,P ＞ 0.05).Conclusions 1.There were distributions of the Trp64Arg polymorphism of β3-AR gene and the C2549A polymorphism of leptin gene in Kazak school-aged children in Xinjiang.2.These results suggested that C2549A mutation of leptin gene might have certain protective effect in Kazak children obesity,and the Trp64Arg polymorphism of β3-AR gene might be associated with Kazak children obesity.3.These results suggested that the Trp64Arg polymorphism of β3-AR gene and the C2549A polymorphism of leptin gene in the same individual joint action might be associated with Kazak children obesity in Xinjiang.

OBJECTIVETo investigate the role of the extracellular signal-regulated protein kinase 1/2 (ERK1/2) pathway in erectile dysfunction (ED) caused by the absence of testosterone (T).

METHODSWe randomly divided 30 eight-week-old healthy male SD rats into groups A (control) , B (castration), and C (castration + androgen replacement). The rats in groups B and C were castrated surgically, and those in C injected with T undecanoate (100 mg/kg) at 1 week after castration, while the others with 0.9% normal saline instead. At 1 month after treatment, we determined the serum T level, intracavernous pressure (ICP), and mean carotid arterial pressure (MAP) of the rats, and detected the expressions of ERK1/2 and endothelial nitric oxide synthase (eNOS) by Western blot.

RESULTSThe serum T level was significantly lower in group B ([1.27 ± 0.48] nmol/L) than in A ([17.14 ± 1.07] nmol/L) and C ([16.24 ± 1.90] nmol/L) (P < 0.05), and so were ICP and MAP (P < 0.05). The expression of ERK1/2 showed no statistically significant differences among the three groups (P > 0.05), that of phosphatase ERK1/2 was markedly higher while that of eNOS remarkably lower in group B than in A and C (both P < 0.05).

CONCLUSIONAndrogen replacement may improve the erectile function of castrated rats by regulating the ERK1/2 pathway.

Androgens ; therapeutic use ; Animals ; Blotting, Western ; Erectile Dysfunction ; drug therapy ; metabolism ; Hormone Replacement Therapy ; MAP Kinase Signaling System ; Male ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Orchiectomy ; Penile Erection ; Penis ; Rats ; Rats, Sprague-Dawley ; Testosterone ; analogs & derivatives ; therapeutic use

Objective:To explore the category,prevalence and related factors of comorbidities associated with Tourette syndrome.Methods:125 patients with TS according to CCMD-3(Chinese Classification of Mental Disorders,3rd edition)were assessed with a self-designed family circumstance questionnaire,YGTSS,CBCL,Leyton obsessive-compulsive scale,and Conner's Child Behavior Checklist.Results:Of 125 TS patients,the comorbidities included attention deficit and hyperactivity disorder(ADHD,41.6%),obsessive-compulsive disorder(OCD,25.6%), anxiety disorders(8.0%),depressive disorders(4.8%),conduct disorders(8.0%),self-injurious behavior(3.2%),and sleep disorder(2.4%).Conclusion:There are many kinds of comorbid disorders at high prevalence in TS patients. These comorbidities adversely influence the therapy and prognosis of TS and are taken as the possible reasons for social function deficit.