Objective The genetic diversity of the natural populations of Gardenia jasminoides were investigated to provide scientific basis for its resources protection and rational utilization. Methods Fourteen pairs of EST-SSR primers were screened in 19 natural populations of 573 individuals to calculate the genetic parameters of G. jasminoides, and further cluster analysis was then carried out. Results Fourteen pairs of EST-SSR primers generated 75 loci, which showed high genetic diversity maintained in natural populations of G. jasminoides (He = 0.703). Mean population gene diversity (Nei) within populations was 0.603, the Shannon’s diversity index (I) was 1.10. Moderate genetic differentiation (Fst= 0.141) and high gene flow (Nm = 1.523) among populations have been showed too. AMOVA analysis revealed that genetic variation within populations was the main sources of total variation. The Mantle test showed there was no significant correlation between genetic distances and geographic distances. Moreover, significant bottlenecks effects in two-phased model of mutation (TPM) test in 73.7% populations were detected in recent history. Conclusion The results in this study indicated that high level genetic diversity were existed in the natural G. jasminoides populations.

Objective: In order to obtain SSR primers which has good universality among the main tree species of Clerodendrum L., and then molecular screening of DNA fingerprints was used in closely related species and genetic research of these ethnical medicinal resources. Methods: An experiment was conducted to study the transferability of 19 pairs of SSR primers developed from C. izuinsulare and C. trichotomum and amplified in nine samples from nine species of Clerodendrum L. Results: Seventeen pairs of 19 primers had the amplification products in C. cyrtophyllum, a cosmopolitan species, the transferability ratio was the highest (89.5%). Seven pairs of primers were successfully amplified in all samples of Clerodendrum L. species, and the transferability ratio was 36.8%. Six pairs of polymorphism SSR primers were used to construct the DNA fingerprints of nine Clerodendrum L. species. In the primers, the CI140 identified seven Clerodendrum L. species except C. fortunatum and C. lindleyi. Coupled with CI132, CT042, CI107, or CI143, they identified all of the nine Clerodendrum L. species. Then, the genetic distances were used to generate a UPGMA tree. The results showed that C. fortunatum, belonging to Ser. Axilliflorae schauer, formed a cluster, and C. mandarinorum and C. cyrtophyllum formed another cluster, while the clusters of the others were difference from the shape classifications. Conclusion: The experiment proves a good way to develop Clerodendrum L. SSR primers from other related species. This study provides the important reference for the use of molecular markers of Clerodendrum L.

Objective To analyze the etiologies,clinical characteristics and prognostic factors of patients with acute renal failure(ARF)admitted to the hospital at high altitude.Method This retrospective study included clinical data of patients with acute renal failure in the General Hospital of Tibet Military Command from May 2001 to April,2006.Results There were 85 male patients and 63 female patients with mean age(42.4?18.1)years old.Among 148 patients with acquired ARF,52.7% was iatrogenic or nosoeomal origin, demonstrating a trend of increasing.The ARF included pre-renal(n=48,32.4%),renal parenchymal(n= 90,60.8%)and post-renal(n=10,6.8%)in origin.Acute high altitude sickness(n=20)was the major causes of pre-renal ARF.Renal parenchymal ARF could be classified into glomerular vascular lesions(n=24), acute tubular necrosis(n=53),acute interstitial nephritides(n=12),and contusion of unitesticle(n=1).of 90 cases of renal parenchymal ARF,39 patients(43.3%)were induced by medicines.Lithiasis was the major causes of post-renal ARF.The mortality of ARF in our study was 42.6%.The mortality of patients contracted ARF in hospital was much higher than that of patients community ARF in community(55.1 vs 23.6%;P=0.01). There was no significant differences of the mortality between the patients with and without dialysis treatment. Univariate analysis showed that prognosis was correlated with age,the presence of hematuria and oliguria or anuria Hb,and the number of organ system failures.The logistic regression showed that age,Hb and the number of organ system dysfunction were the predictors of mortality.Conlusions The major causes of ARF at high altitude were acute high altitude sickness and the use of medicines with nephrotoxicity.The morbility and mortality of nosocomisl ARF increased significantly.Prevention of MODS is a key management to decrease mortality in severe ARF.

To evaluate the absorptive characteristics of furfural onto biomass charcoals derived from rice husk pyrolysis, we studied the information of the structure and surface chemistry properties of the rice husk charcoals modified by thermal treatment under nitrogen and carbon dioxide flow and adsorption mechanism of furfural. The modified samples are labeled as RH-N2 and RH-CO2. Fresh rice husk charcoal sample (RH-450) and modified samples were characterized by elemental analysis, nitrogen adsorption-desorption isotherms, Fourier-transform infrared spectroscopy and Boehm titration. The results show that fresh rice husk charcoal obtained at 450 degrees C had a large number of organic groups on its surface and poor pore structure. After the modification under nitrogen and carbon dioxide flow, oxygenic organics in rice husk charcoals decompose further, leading to the reduction of acidic functional groups on charcoals surface, and the increase of the pyrone structures of the basic groups. Meanwhile, pore structure was improved significantly and the surface area was increased, especially for the micropores. This resulted in the increase of π-π dispersion between the surfaces of rice husk charcoals and furfural molecular. With making comprehensive consideration of π-π dispersion and pore structure, the best removal efficiency of furfural was obtained by rice husk charcoal modified under carbon dioxide flow.
Adsorption ; Biomass ; Carbon Dioxide ; Charcoal ; Furaldehyde ; chemistry ; Nitrogen ; Oryza ; Spectroscopy, Fourier Transform Infrared ; Surface Properties

AIM: To construct prokaryotic expression vector of His-tagged human IP-10 for further study of its biological function in the inflammatory response. METHODS: The coding sequence of IP-10 lacking signal peptide was amplified from human lung cDNA library by polymerase chain reaction (PCR) and the fragment was cloned into pET-14b plasmid for the construction of His-tagged fusion protein expressing vector, pET-14b/IP-10. After being identified by enzyme digestion and sequencing, the recombinant vector was transformed into a strain of E. coli, BL21 (DE_3). The expression of His-tagged fusion protein was induced with IPTG and purified with Ni+-NTA affinity chromatography. Then the chemotactic activity of IP-10 was determined by transwell migration assay on THP-1 cells. RESULTS: The construction of pET-14b/IP-10 recombinant vector was proved by enzyme digestion and sequencing. The fusion protein IP-10, which was purified by a routine Ni+ affinity method, had an activity on the induction of cell migration of THP-1. CONCLUSION: We successfully construct IP-10 fusion protein expressing vector and get the fusion protein with high bioactivity, which provides essential materials for the future studies on IP-10.

Objective To construct the vector of mouse HMGB1 promoter-driven red fluorescent protein reporter gene so as to supply a tool for the study on the expression regulation of HMGB1 gene in mammalian cells and related signal transduction mechanism. Methods The mouse HMGB1 promoter sequence was subcloned into a red fluorescent protein reporter gene vector, pDsRed1-1. The recombinant vector pDsRed1-1-HMGB1p was then transfected into NIH3T3 cells by liposome, and the intracellular activity of HMGB1 promoter was observed under a fluorescence microscope in normal condition or after tumor necrosis factor-α (TNF-α) stimulation. Results The recombinant plasmid was confirmed by enzyme digestion and DNA sequence analysis. The vector was expressed with red fluorescence at a low level in the rest NIH3T3 cells, but the expression was highly increased by the stimulation with TNF-α. Conclusion A red fluorescent protein reporter gene vector driven by mouse HMGBI promoter is constructed successfully, which can be expressed in mammalian cells with a physiological response to TNF-α stimulation, thus providing an important and convenient tool for the study on the regulatory mechanisms of HMGB 1 gene expression.

Objective To explore the clinical outcome of improved technique of Gamma nail in the treatment of intertrochanteric hip fracture of the elderly patients. Methods From March 2002 to October 2006.39 patients with intertrochanteric hip fracture were operated by improved technique of Gam-ma nail.There were 18 males and 21 females at average age of75.7 years(67_98 years).There were 6 patients with type A1 fracture,24 with type A2 fracture and 9 with type A3 fracture according to AO/ASIF classification.Of all.36 patients(92.3%)had osteoporosis.The operation improvements included the following points:(1)The patients were placed at the lateral decubitus position with the fractured limb on the uppermost,with flexion of knee and hip of 60°.The normal hip and knee were flexed as possible.(2)One-off indirect traction reduction was used after general anesthesia. no requirement of continuous mechanical traction.(3)C-arm image intensifier was employed to obtain normal and lateral projections.Results Of all,35 patients were followed up for a mean period of3 years and 2 months, ranging from 6 months to 5 years and 2 months.Operation data showed incision length of(4.3±1.2)cm,mean opera-tion time of(46±10)minutes,intraoperative bleeding volume of(65±26)ml and intraoperative X-ray exposure of(3.0±2.1)times.Postoperative recovery data showed survival in one-year follow up,with ambulation time of(10.5±3.6)days and fracture union time of(10.9±2.1)weeks.Mean Parker's score wag(6.9±3.2)points 6 months after operation. Conclusions Improved technique of Gamma nail can shorten operation duration,reduce operative trauma and bleeding,reduce X-ray exposure and im-prove success rate of surgery.as facilitates early pest-operative recovery and reduces the perioperative mortality rate of the elderly.

Objective To explore the importance and methods of retaining articularis during pri- mary total hip arthroplasty(THA)and reconstruct soft tissue balance of hip joint after THA.Methods From February 2003 to August 2005,41 eases(43 hips)including 19 males and 22 females at age of 46- 80 years(mean 66.5 years)were treated with THA with retained capsule(Group R)and other 42 cases (44 hips)including 20 males and 22 females at age of 43-80 years(mean 64.3 years)with standard THA (Group S).Preoperative diagnosis found femoral neck fractures(GardenⅢⅣ)in 13 cases(13 hips)in Group R and 14(14 hips)in Group S;acetabular dysplasia(CroweⅢ)in 9(9 hips)in Group R and 8 (hips)in Group S;Osteoarthritis in 6(8 hips)in Group R and 7(8 hips)in Group S;and femoral head osteonecrosis(FicatⅢⅣ)in 13(13 hips)in Group R and 13(14 hips)in Group S.There were 13 hips of cement prostheses in Group R and 11 in Group S,8 cementless prostheses in Group R and 8 in Group S, 22 cement and cementless prostheses in Group R and 23 in Group S.Gibson's approach was used in both groups.Group R used the method of retaining capsule and little supination muscles during the operation to reconstruct responsibly soft tissue balancing of postoperation for THA.For comparison,Group S used the method of standard which resected a lots of capsule and didn't reconstruct it.The comparative items between Group R and Group S included incisional length,operative time,operative bleeding,drainage transfusion, infection,dislocation,postoperation standing,postoperation walking and Harris's score.Results All cases in Group R and Group S were followed for 6-22 months(mean 16.5 months in Group R and 16.7 months in Group S).There was significantly statistical difference upon interoperative and postoperative data between Group R and Group S.The result of Group R was significantly better than that of GS.Conclu- sion Retaining articularis during primary THA can minimize operative trauma,reconstruct soft tissue bal- ance and augment hip stability to get postoperative functional recovery.

OBJECTIVETo construct a recombinant adenovirus vector for expressing interferon-gamma-inducible protein 10 (IP-10) by homogenous bacterial recombination.

METHODSIP-10 gene was cloned into the shuttle plasmid pAdTrack-CMV that contained the coding sequence of enhanced green fluorescent protein (EGFP). The shuttle plasmid was then transformed into E. coli BJ5183 with pAdEasy-1 vector by chemical transformation. The recombinant adenovirus vector pAd/IP-10 was identified by enzyme digestion with Pac I and the linearized plasmid was transfected into HEK293 cells.

RESULTSThe positive clones were identified with enzyme digestion and polymerase chain reaction (PCR) and were further verified by DNA sequencing. The recombinant adenovirus of high titration was obtained after transfection and packaging in HEK293 cells.

CONCLUSIONA recombinant adenovirus vector for expression of IP-10 has been constructed successfully and high-titer active adenovirus is obtained for functional study of IP-10 protein.

Adenoviridae ; genetics ; Cell Line ; Chemokine CXCL10 ; genetics ; metabolism ; Cloning, Molecular ; Defective Viruses ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Virus Cultivation ; methods

OBJECTIVETo explore the relationship between the mutations of epidermal growth factor receptor (EGFR) gene and clinicopathological characteristics in patients with non-small cell lung cancers (NSCLC).

METHODSParaffin-embedded tissue specimens were obtained from 1444 patients with NSCLC. The genomic DNA was extracted. Mutations of EGFR gene (exons 19 and 21) were detected by real-time PCR.

RESULTSDNA was available in 1410 cases. Somatic mutations of the EGFR gene were identified in 401 cases (27.8%). Among patients with EGFR mutations, 41.4% (n=166) had del E746-A750 of exon19, 6.7% (n=27) had del L747-P753insS of exon 19, 50.3% (n=201) had L858R of exon 21, and 1.5% (n=6) had L861Q of exon 21. Woman, non-smoker and adenocarcinoma showed a higher percentage of EGFR mutation (43.2%, 37.6%, and 33.5%, respectively). However, there was no association among age, grades, lymph node metastasis, and TNM stages (P>0.05). The mutation rate of BAC subtype (61.3%, 19/31) and adenocarcinoma with BAC features (48.0%, 12/25) was significantly higher than that of conventional adenocarcinoma (32.4%, 336/1038). A further assess of the smoking status found a trend that the more increased smoking exposure, the lower the incidence of EGFR mutations. A multivariable analysis revealed that adenocarcinoma, never smoking, and female were independently associated with EGFR mutations (odds rations=3.381, 2.393, and 1.727, respectively).

CONCLUSIONSThe detection rate of EGFR mutation is higher in Chinese patients, especially in non-smoking female patients with adenocarcinoma. Real-time PCR is a sensitive and accurate method to detect the mutations of EGFR gene and can therefore provide useful information for clinical treatment.

Adenocarcinoma ; genetics ; Adenocarcinoma, Bronchiolo-Alveolar ; genetics ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; Exons ; Female ; Genes, erbB-1 ; genetics ; Humans ; Lung Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; Mutation ; Mutation Rate ; Real-Time Polymerase Chain Reaction ; Receptor, Epidermal Growth Factor ; genetics ; Sex Factors ; Smoking ; Young Adult