Objective To establish the specific 16S 23S rRNA gene spacer regions map of different bacteria by PCR, RFLP(restriction fragment length polymorphism ),DNA clone and sequences analysis. Methods A pair of primer was selected from highly conserved sequences adjacent to the 16S 23S rRNA spacer region. The farget rRNA regions from 61 strains of standard bacteria and corresponding clinical isolates representing for 20 genera and 26 species were amplified by PCR,and thereafter analyzed RFLP, DNA clone and sequences analysis.Meanwhile, all the specimens were examined by bacterial culture and PCR RFLP analysis. Results 26 different standard strains presented one band,two bands,three bands and more than three bands respectively, the sensitivity of which reached 2.5 CFU and had no cross reaction to the human genomic DNA,fungus and virus.14 species could be distinguished immediately by PCR, other 10 species must be identified by further Hinf I or Alu I digestion. K.pneumoniae and E.durans differentiate only at the site of 779 th nucleotide according to the sequence analysis, and only one enzyme Xma III could discriminate them.15 specimens from 42 septicemic neonates were blood culture positive and the positive rate was 35.7%. However, 27 specimens were positive by PCR and the positive rate was 64.2%,which was significantly higher than that of the blood culture( P

Objective To discuss the feasibility,safety,technical points and clinical effects of completely thoracoscopic resection of benign neurogenic tumors on apical chest.Methods From January 2004 to June 2009,11 patients underwent surgical resection of benign neurogenic tumours on apical chest.A complete thoracoscopy was used in 5 cases,and the remaining 6 cases received traditional open thoracotomy.By analysis on the clinical symptoms,tumor types,complications,operative time,blood loss and drainage time after operation,the advantages and disadvantages of complete thoracoscopy were compared to traditional open thoracotomy for resection of benign neurogenic tumors on apical chest.Results There was one patient in each group that suffered from light transient Horner's syndrome,who recovered spontaneously.The group of complete thoracoscopy was superior to the group of traditional open thoracotomy in operative time,blood loss during the operation,drainage time and postoperative hospital stay.Conclusion For benign neurogenic tumors on apical chest,a resection with complete thoracoscopy is as safe and effective as the traditional open thoracotomy,and the former is characterized by less operative trauma and quicker recovery.

Objective To evaluate the efficacy of self-retaining suture (QuillTM SRS) in retroperitoneal laparoscopic partial nephrectomy for complicated renal tumor by assessing perioperative parameters.Methods Between 2010 and 2012,78 cases of complicated renal tumor (R.E.N.A.L score ≥ 7) treated by retroperitoneal laparoscopic partial nephrectomy (LPN) with two layers continuous knotless barbed suture (QuillTM SRS group) (n=30) or traditional absorbable vicyl suture (non-SRS group) (n=48) were retrospectively analyzed.In QuillTM SRS group,2-0 Quill SRS was used to suture the deep wound bed,and the second outcr layer renorrhaphy was performed with a 1-0 Quill SRS by the same way.In non-SRS group,the inner layer was sutured using a 15cm in length 2-0 monicryl suture by the same method mentioned above.A second outer layer was sutured with 1-0 vicryl suture across the wound.Cases were matched for R.E.N.A.L score.Comparison was made in term of operation time,preoperative parameter and perioperative complications between SRS group and non-SRS group.Results Renorrhaphy was successfully performed in all cases except 1 case converting to open surgery in non-SRS group.Mean warm ischemia time in SRS group was shorter than non-SRS group (18 vs 25 min,P =0.021).The proportion of bleeding requiring intervention in the non-SRS group (7/48,14.5％) was 4.3-fold higher than that of the SRS group (1/30,3.3％),but the differernce is not significant (P＞0.05).There were no significant differences between two groups in postoperative creatinine changes.Limitations of this study include the absence of randomization and the relative small sample size.Conclusions SRS can be safely used for complicated renal tumor during LPN,and SRS can significantly reduce the WIT and may also reduce bleeding during the operation.

ObjectiveTo report the results of preventive control program of severe thalassemias in Zhuhai City of Guangdong Province from 1998 to 2010.MethodsAs the guide centre of marriage and childbearing and the greatest maternity hospital in Zhuhai City of Guangdong Province,Zhuhai Municipal Maternity and Child Healthcare Hospital constructed the genetic screening network for thalassemias testing and referred for follow-up and for genetic counseling.The couples for premarital medical examination or regular healthcare examination in pregnancy were enrolled to this preventive control program.A conventional strategy of screening for heterozygote was used to identify the α- and β-thalassemia traits in women and their spouses according to the standard procedures of hematological phenotype analysis which was recommended by Thalassemia International Federation (T IF).Then those suspected couples at risk were diagnosed for α- and β-thalassemia by PCR-based DNA assays.The couples at risk for severe thalassemias were counseled and offered prenatal diagnosis and termination of pregnancy in case of an affected fetus in the rights of consent and of option voluntarily.ResultsFrom January 1998 to December 2010,85 522 brides and grooms-to-be for premarital screening and 41 503 pregnant women in addition to 14 141 partners for prenatal screening were recorded,the covering rates of premarital screening and prenatal screening in the city were 92.698％ (from 1998 to 2003) and 27.667％ (from 2004 to 2010),respectively.Totally 10 726 cases were found to be the carriers of thalassemias,with 7393 for o-thalassemia (5.237％,7 393/141 166) and 3333 for β-thalassemia (2.361％,3 333/141 166).A total of 257 couples at-risk for severe thalassemias were detected including 190 for α-thalassemia and 67 for β-thalassemia.Among them,251 (97.7％,251/257) couples were performed prenatal diagnosis.During the preventive control program,a total of 72 fetuses with severe thalassemias including hemoglobin H disease were voluntarily terminated.In Zhuhai City,the average annual birth rate of fetuses with severe thalassemia was declined by 32.9％ (49/149).ConclusionsThis study has reduced effectively birth rate of perinatal infants with severe thalassemias in Zhuhai City by genetic screening and prenatal diagnosis of thalassemia in the large population of 13 years.Our summary comes out of technical proposals for prenatal screening and diagnosis,which could be take example by preventative control of thalassemia in other regions of China where are prevalent.

BACKGROUND: Wnt signaling pathway plays an important regulative role in the embryonic development processes. Accordingly, it is of great significance to establish the cell model of Wnt signaling pathway so as to conduct study on it. OBJECTIVE: To establish Wnt signaling pathway cell model by transfecting L-M (TK-) cells with Wnt3a eukaryotic expression plasmid, and to investigate the effect of canonical Wnt signal pathway on the β-catenin subcellular distribution. METHODS: The eukaryotic expression plasmid pgk-Wnt3a-pcDNA3.0 after amplification was digested by restriction endonuclease first. Then it was transfected together with the control plasmid pgk-neo-pcDNA3.0 into L-M (TK-) cells via lipofection, after which the cell colony was screened by G418 for amplification. RT-PCR was used for detecting the expression products and the indirect immunofluorescence assay for observing the effect of Wnt3a on the β-catenin subcellular localization of L-M (TK-) cells. RESULTS AND CONCLUSION: The Wnt3a plasmid was verified by endonuclease digestion to have produced the expected plasmids after amplification. According to the RT-PCR detection to the 10 stably-transfected cell colonies achieved by 3 weeks of G418 screening, it was seen, on the L-Wnt3a cDNA, a strip of bright band of 320 bp in length, which showed that the products of amplification were exactly the expected fragments and that the Wnt3a plasmid was expressed on mRNA transcriptional level after being transfected with L-M (TK-) cells. In contrast, no expected band was found on the cDNA of L-M (TK-) calls transfecting the control plasmid. In addition, the immunofiuorescence assay detection showed that the protein expression of Wnt3a was found in the cytoplasm of the L-M(TK-) cells tranfecting Wnt3a plasmid, while for those transfecting the control plasmid, it was opposite. β-catenin, as showing by bright red fluorescence, was found to concentrate and enter into the nucleus of the L-M (TK-) cells transfecting Wnt3a plasmid, while for those transfecting the control plasmid, it was opposite. Cell model with continually activated Wnt signaling pathway is established. The stable expression of Wnt3a in L-M (TK-) cells transfected with pgk-Wnt3a-pcDNA3.0 is obtained. The expression of Wnt3a is able to promote the transfer of β-catenin from cytoplasms into nucleus in L-M (TK-) cells.

Objective To explore the effect of luteolin on the proliferation of osteosarcoma stem cells.Methods CD133 +osteosarcoma stem cells were separated from MG63 cells by flow cytometer.MTT was used to investigate the effects of luteolin(0,0.01,0.02,0.04 mg/mL)on the proliferation of osteosarcoma stem cells.Western blot was used to detect the levels of Ki67 protein and components of JAK2/STAT3 signal pathway in osteosarcoma stem cells induced.Results After sorting,the content of the CD133 +fraction was enriched up to(87.60 ±5.06)%.MTT assay showed that,compared with the control group,luteolin(0.01,0.02,0.04 mg/mL)inhibited proliferation of CD133 + osteosarcoma stem cells(P <0.05).Western blot also showed that luteolin significantly decreased the level of Ki67 compared with the control group(P<0.05).In addition,the luteolin inhibited the expression of p-JAK2 and p-STAT3 in JAK2/STAT3 signal pathway of CD133 + osteosarcoma stem cells compared with the control group ( P <0.05 ) . Conclusion Luteolin might be a suppressor of osteosarcoma stem cells.

OBJECTIVE Oxidative sress is one of the key factor responsible for occurrence and development of hepatic fibrosis, a common consequence of chronic liver injury of multiple etiology. Nuclear factor erythroid 2-related factor 2 (Nrf2) serves as a major regulator of a celular defense system against oxidative stress. Xiaochaihutang (XCHT), a compound of seven botanical extracts used for liver diseases traditionally in East Asia. However, few studies have investigated its anti-hepatic fibrosis effects and pathophysiological mechanism of action. The present study was designed to confirm the anti-hepatic fibrosis effects and explore its potential mechanism of action by investigating the intervention of Nrf2 pathway. METHODS Liver fibrosis was induced by repeated injection of Carbon tetrachloride (CCl4) over a period of 9 weeks. Starting from the 6th week, the animals in treatment groups were given the appropriate dose of XCHT granules and silybin. Biochemical parameters, histological changes of the liver and alpha-smooth muscle actin (α-SMA) were determined. The expressions of Nrf2, Keap1, Nqo1, HO-1, Gclc and Gclm were assessed by RT-PCR and Western blot. RESULTS CCl4 caused a significant fibrosis damage in the rat liver and the liver functions and fibrosis degree were significantly improved by XCHT (5 g·kg- 1 and 10 g·kg- 1). XCHT (5 g·kg- 1 and 10 g·kg- 1) treatment significantly decreased the number of cells labeled with α-SMA antibodies. Moreover, XCHT (5 g·kg-1 and 10 g·kg-1) significantly increase Nqo1, HO-1, Gclc and Gclm expressions in the liver. CONCLUSION These studies establish XCHT is a potentially useful therapeutic agent for treatment of hepatic fibrosis and it might be via regulation of Nrf2 pathway in rats against oxidative stress, making further efforts to inhibiting the activated HSCs. Activation or up-regulation of Nrf2 pathway may be an alternative treatment strategy for liver fibrosis.

Notopterygium incisum is the important medicinal materials of the Tibetan-Qiang medical system in China, also one of the rare and endangered medicinal materials in the Plateau areas in the meantime. Taking the planting of in Sichuan province as an example, research on the N. incisum in Sichuan utilize remote sensing and GIS techniques, bind growth environment factor, including height factor, average annual precipitation, average annual temperature, forest information, were chosen according to habitat conditions. And combine field measurement to verify. The results indicate that N. incisum resources in Sichuan province were mainly distributed in the alpine valley and the northwest of the plateau, which suitability distribution areas of 4145 km2 approximately and accounting for 2% of the total area. Suitability areas accounting for more than 2% of the respective total area in Heishui county, Lixian county, Xiaojin county, Kangding county, ect. According to the field investigation and the related document information record, drawn that the suitability distribution based on RS and GIS were corresponded with the actual distribution areas of N. incisum resources. It's feasible to divide the suitability distribution area of N. incisum using RS and GIS, which will provide a scientific basis for a comprehensive investigation of the distribution as well as its rational exploitation and protection.
Apiaceae ; China ; Conservation of Natural Resources ; Geographic Information Systems ; Telemetry

OBJECTIVEWith the objective of discovering novel putative chromosomal regions and special genes involved in the carcinogenesis, progression and metastasis of laryngeal squamous cell cancer (LSCC).

METHODSDNA copy profile of LSCC were obtained and analyzed by comparative genomic hybridization (CGH) and a computerized digital image analysis system. cDNA microarray of LSCC was performed and the profile was analyzed by Hierarchical clustering.

RESULTSCGH analysis showed average-12.9 gains and losses of chromosomes in LSCC. Relatively high frequencies of gains were found at 3q15-21 (14/18), 5p12-13 (11/18), 8q22-24 (6/18), 11q12-13 (8/18), 15q21-23 (7/18) and 18p11 (8/18), while those of losses at 1p13-21 (8/18), 3p21-23 (14/18), 5q21-22 (14/18), 9p12-pter (11/18) and 13q21-31 (8/18). Hierarchical clustering analysis showed that the differentially expressed genes were segregated into three groups. Three genes differentially expressed in process I (normal tissue to cancer) and process II (cancer to lymph node metastasis), and the Cy5/Cy3 ratios of twelve genes were either higher than 5.0 or lower than 0.2 in process I or process II. The fifteen special genes were first reported possibly to be the relationships with LSCC. In particular, 4 genes of them, which were cytochrome C oxidase Va, PPBP, EPHX2 and PON1, were first reported to correlate with tumorigenesis. SH3GL2, which was one of the 15 special genes, was located at one of the special chromosome regions, 9p12-pter.

CONCLUSIONThe important genes and special chromosomal aberrances might provide us a clue for further investigation of carcinogenesis, progression and metastasis in LSCC.

Adult ; Aged ; Carcinoma, Squamous Cell ; genetics ; pathology ; Chromosome Aberrations ; DNA, Neoplasm ; analysis ; Disease Progression ; Female ; Gene Expression ; Humans ; Karyotyping ; Laryngeal Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Nucleic Acid Hybridization ; Oligonucleotide Array Sequence Analysis

OBJECTIVEAn improved tracheal intubation method was used in preparation of myocardial infarction model in the mouse for increasing the success rate.

METHODSThe mice were intubated by an improved trachea cannula through oral cavity. The left anterior thoracotomy performed. The myocardial infarction model was made by legating the left anterior descending coronary artery in mouse. The color of heart was observed, and electrocardiogram was recorded. The survival rate and pathologic change were observed after two weeks of operation.

RESULTS40 myocardial infarction model mice were made by improved trachea cannula. The color of ventricles anterior wall had got madder red, and ST stages were raised on II leads of electrocardiogram in all of model mice. After two weeks, 27 mice were survival. The survival rate was 87.1% except for accidental death during operation. The heart chamber expanded and ventricular wall became thin in myocardial infarction mice by eyes. After pathological sections were stained, by HE cardiac muscle fibers ruptured or lysed. There were some of necrosis of myocardiac cells and many of infiltration of inflammatory cells.

CONCLUSIONApplication of an improved tracheal intubation method simplified operation of tracheal intubation during preparation of myocardial infarction model in the mouse. The trauma was tinier than the other one, and achievement ratio of the model preparation was improved.

Animals ; Disease Models, Animal ; Intubation, Intratracheal ; methods ; Male ; Mice ; Myocardial Infarction