Millettia pinnata L. is a leguminous tree with great potential in biodiesel applications and also a typical semi-mangrove. In this review, we presented several aspects about the recent research progress in molecular biology of M. pinnata. We descrived several types of molecular markers used to assess the genetic diversity and phylogeny of this species, genome and transcriptome analyses based on high-throughput sequencing platform accomplished for this species, and several gene and genomic sequences of this species isolated for further research. Finally, based on the current research progress, we proposed some orientations for future molecular biology research on M. pinnata.
Base Sequence ; Gene Expression Profiling ; Genetic Variation ; Genome, Plant ; Genomics ; Millettia ; genetics ; Phylogeny ; Trees ; genetics

OBJECTIVE To investigate the bacterial resistance of clinical isolates from pediatric hospital for the guidance of rational use of antibiotics.METHODS Disc diffusion test(Kirby-Bauer method) was used to study the antimicrobial resistance(fastidious bacteria were detected by E test).WHONET5 was applied for analysis.(RESULTS) In the period of study from 2002 to 2003,2 303 strains which were the first isolated from each patient were collected.Of 2 303 clinical isolates,Gram positive organisms accounted for 29.7%,Gram negative ones for 70.3%.Escherichia coli,Klebsiella spp,Staphylococcus aureus,coagulase-negative staphylococci and Pseudomonas aeruginosa were the most common strains among the isolates.Meticillin-resistant S.aureus(MRSA) and meticillin-resistant coagulase(-negative) staphylococci(MRCNS) accounted for 9.7% and 67.6% of S.aureus and coagulase-negative(staphylococci),respectively.Resistant rates of MRSA and MRCNS were higher than that of meticillin-susceptible S.aureus(MSSA) and meticillin-susceptible coagulase-negative staphylococci(MSCNS) to antimicrobial agents commonly used in clinic.No vancomycin resistant strains of staphylococci were found. 4.1% of Enterococcus spp were vancomycin resistant strains.Resistant rate of Streptococcus pneumoniae was 11.9% to penicillin. Most of isolates of Enterobacteriaceae were susceptible to imipenem.The incidences of E.coli and Klebsiella spp producing extended spectrum beta-lactamases(ESBLs) isolates were 49.7% and 63.1%,respectively.The resistance rates of(ESBLs) producing strains to antimicrobial agents(except carbapenems) were higher than those of ESBLs nonproducing ones.CONCLUSIONS Bacterial resistance is still or even a more serious clinical problem than before.The(surveillance) of antimicrobial susceptibility in pediatric hospital is of great significance.It is also very important to promote the rational use of antimicrobial agents so that resistance is minimized and take effective strategy for the control of the problem.

Objective To construct and identify recombinant expression plasmid of small interfering RNA (siRNA)targeting hepatitis B virus X protein(HBx), and observe its effect on mitoehondrial function in healthy liver cell line steadily expressed HBx gene (HL-7702/HBx). Methods Two siRNA sequences containing short hairpin structure, which target on the total length HBx gene, were synthesized and cloned into the vector psiRNA-Hh1GFPzeo to eonstruct recombinant expression plasmids pX1 and pX2. Non-specific recombinant pScr plasmid served as control. After siRNA transfected into HL-7702/HBx cells line by liposome, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were performed to identify the suppressive effect on HBx expression. Levels of intraeellular reactive oxygen species (ROS) and mitochondrial membrane potential (△ m) were determined by flow cytometry. The experimental results were compared by analysis of variance. Results Successful constructions of pX1 and pX2 were confirmed by restriction enzyme digestion and sequencing. The expressions of HBx mRNA and protein after 48 h of transfection into HL-7702/HBx cells in control group were 0.65± 0.12 and 0.62± 0.09, respectively, which were both higher than those (0.33±0.10 and 0.19±0.08, respectively) in group pX1 (t=4.73, P<0.05; t=7.53, P<0.05) and those (0.48±0.10 and 0.37±0.11, respectively) in group pX2 (t=2.39, P<0.05;t=4.43,P<0.05). But the inhibition of group pX1 was stronger than that of pX2 (t=2.28,P<0.05). Levels of ROS and △ m after RNA interference were 5.00±0.38 and 33.86±0.50, respectively, while those in control group were 72. 10±0. 55 and 3. 57±0.26, respectively (ROS: t=276.22, P<0.05; △ m: t=107.15, P<0.05). Conclusions siRNA targeting HBx can efficiently and specifically suppress the HBx expression in HL-7702/HBx cells, and decrease the level of ROS and increase the level of △ m, thus relieve cellular oxidative stress.

Objective To investigate the feasibility of restoration of cartilage defect with silk fibrin/chitosan(SF-CS)biological scaffold compound by induced bone marrow mesenchymal stem cells (BMSCs)in the elderly rabbits.Methods BMSCs were extracted,cultured and induced to differentiate,then inoculated into SF-CS three-dimensional scaffold restoration.54 rabbits(aged 16-18months)were divided into scaffold restoration,single scaffold and control groups(n=18 per group).The right knee joint was used for building cartilage defect model and implanted by scaffolds.General observation,tissue staining and modified Wakitani histological scoring were performed at 4,8 and 12weeks after operation.Results SF-CS scaffold was structured by multiple interlinked pores.The average pore size was 151.72 μm.The porosity was(92.72±4.78)％.The imbibition rate was (141.10± 6.87)％.BMSCs was grown well and proliferated dynamically in SF-CS scaffold after induction.At 12 weeks,the cartilage defect was basically repaired,type Ⅱ collagen was positively expressed and the scaffold was almost assimilated in scaffold restoration group.In single scaffold group,the cartilage defect was repaired mainly by fiber tissue,type Ⅱ collagen was less expressed and the scaffold almost degraded while the cartilage defect was repaired badly in control group.The scaffold restoration group was superior to single scaffold and control groups(P＜0.05)in improving the Wakitani score.Conclusions The SF-CS scaffold as BMSCs carrier may restore cartilage defect in knee joint of the elderly rabbits.

Objective To establish a HPLC method for the determination of effective components in Radix Notoginseng in San Xiong Zhitong Ointment.Methods Kromasil KR100-5 C18 column(250 mm?4.6 mm,5 ?m)and gradient elution were used.Solvent A was acetonitrile and solvent B was water.The detection wavelength was set at 203 nm.Results The four components were isolated well.The linearity was fine with the recoveries of 98 %～100 %.Conclusion The quantitative method for determining the ingredient of San Xiong Zhitong Ointment is simple,feasible and repeatable,and is beneficial for quality control of San Xiong Zhitong Ointment.

Objective To establish a method of HPLC assay for determining stilbene glucoside in Zishen Ningshen Pills(ZNP),and to study the influence factors on the content of stilbene glucoside in the process of preparation.Methods HPLC was used for the determination of stilbene glucoside in ZNP.Through simulation the process of preparation,the stilbene glucoside content in the intermediate products was determined by HPLC,and its retention rate and metastasis rate were also investigated.Results The resolution and the linearity of stilbene glucoside were fine,the average recoveries being 98 % ～ 102 %.The retention rate of stilbene glucoside in the drying powder was 60.3 %,lower than that in the original medicinal powder.Conclusion The quantitative method for determining the ingredients in ZNP is simple,feasible and reproducible,and is beneficial for quality control of ZNP.The drying process under normal pressure is the main influence factors of the decrease of stilbene glucoside content,and the decompression drying can be taken into account to take the place of the atmospheric drying.

BACKGROUND:Silk fibroin/chitosan/nano hydroxyapatite (SF/CS/nHA) composite scaffold constructed in preliminary experiments has good physical and chemical properties.
OBJECTIVE:To study the capacity and mechanism of SF/CS/nHA composite scaffold for repair of rabbit radial bone defects.
METHODS:Thirty-six New Zealand white rabbits were selected to make animal models of right radial bone defects, and then randomly divided into SF/CS/nHA group, SF/CS group and blank control group. Blank control group had no treatment after modeling. X-ray radiography, gross observation and histopathological observation were performed at 4, 8, 12, 16 weeks postoperatively.
RESULTS AND CONCLUSION:Sixteen weeks after surgery, bone defects in the SF/CS/nHA group were completed replaced by normal bone tissue on X-ray images, and the bone marrow cavity showed complete recanalization with new bone formation;hematoxylin-eosin staining showed bone trabecula and many fusiform bone cells. In the SF/CS group, the bone mineral density in the defect area was slightly lower than that of the normal bone tissues, the bone marrow cavity was partly rehabilitated, and many chondrocytes were seen around bone cells that arranged irregularly with no bone trabecula or bone lamel a. In the blank control group, the images of bone calcification were consistent with normal bone tissues, and a closed bone ununion was formed at each end;hematoxylin-eosin staining showed that the blank control group was fil ed by fibrous connective tissue and a smal amount of bone-like tissues. SF/CS/nHA composite scaffold is better for repair of rabbit radial bone defects.

Objective To explore the expression of chemerin and chemerin receptor ( chemokine-like receptor 1, CMKLR1) during different periods of non-alcoholic fatty liver disease ( NAFLD) rat model induced by methionine- and choline-deficient ( MCD) diet. Methods Thirty-six Wistar rats were divided into control group and MCD group in random. After one week quarantine and acclimation period, these two groups were fed either normal chow or MCD diet. The animals were respectively sacrificed at the first week, the forth week, and the tenth week. The levels of alanine transaminase (ALT), blood lipid profile, liver function, and the content of triglyceride in liver were detected. HE staining was done to observe the morphologic change of liver. The mRNA expression changes of chemerin and CMKLR1 in liver were measured using real-time PCR, and the change in chemerin mRNA level was further confirmed in liver by Northern blot. Finally, the concentration of chemerin in serum was measured by Western blot. Results The mRNA level of chemerin decreased significantly after four and ten weeks MCD feeding, although no obvious changes were found at first week, similar changes were found in serum chemerin (1.00±0.11 vs 0.96±0.39; 1.00±0.12 vs 0.21 ±0.77; 1.00±0.42 vs 0.21 ±0. 11). Contrasting with the change of chemerin(1.00±0.08 vs 0.72±0.10;1.00±0.24 vs 0.63±0. 31 ;1.00±0.05 vs 0.50±0.13), the mRNA level of CMKLR1 increased after MCD feeding( 1.00±0. 14 vs 0. 84±0. 26; 1.00±0. 38 vs 1. 51 ±0. 33; 1. 01 ±0. 13 vs 1. 84 ± 0. 39 ). Conclusion The change of chemerin and its receptor may participate in the process of the nonalcoholic fatty liver disease.

To screen the siRNAs (small interference RNA sequences ) which specifically inhibit the gene expression of TLR2 in human monocyte-derived macrophage , and discuss their prospects on the treatment of HIV at the level of molecular immunology.Methods:We obtained the mRNA sequences of human TLR 2 gene from NCBI gene bank ,then designed three siRNAs by siDESIGNTM software.The siRNA targeting human housekeeping gene GAPDH was used as positive control.The fluorescent labeling missense siRNA sequences (NC-FAM ) was used as negative control.We collected fresh peripheral blood from healthy volunteers and isolated mononuclear cells from the blood samples.The human mononuclear macrophages were then purified from mononuclear cells by utilizing adherence method.Cationic liposome reagent Lipofectamine 2000 was used to mediate siRNAs into the human mononuclear macrophages.The levels of TLR2 mRNA expression of siRNA-transfected monocyte-derived macrophage were determined by q-PCR.Expression of TLR2 protein was determined by Western blot.Results: At 72 h after transfection ,we found that the expression of GAPDH mRNA and protein in positive control group decreased significantly.Also found there existed significant differences between each siRNA group (F=41.957,P<0.001).Compared with negative control ,the relative expression of TLR2 mRNA in all siRNAs groups decreased significantly (P<0.05 ) , and the inhibition rates were 46%, 43%, 43% by three miRNAs respectively.Western blot showed that the expression of TLR 2 protein in siRNAs groups decreased significantly compared with that of control (P<0.05 ).Conclusion: The designed siRNAs in this study could inhibit the expression of TLR 2 gene in human monocyte-derived macrophage ,indicating that mediation of TLR-2 expression by siRNA might be a novel strategy for HIV treatment from the per-spective of molecular immunology.

Objective: Evaluate GII.4 norovirus infection and blocking effects of serum antibodies against HBGAs binding to GII.4 norovirus of population in oyster culture area, provide references for screening of fully human monoclonal antibody. Methods: Using a random survey method to collect blood and saliva samples in oyster culture area, select serum samples from the inland region of Guangdong as control group. Identification of salivary HBGA receptor phenotype and detection of serum antibody levels between two areas by ELISA. A vitro neutralization model was to determine the efficiency of serum antibodies blocking GII.4 norovirus and HBGA receptors binding. Results: The age were (50.68 ± 15.17), (52.52 ± 15.90) and (51.37 ± 13.32) years old of 2015, 2016 in experimental group, and in control group, respectively. Males accounted for 5.9% (70/195), 36.6%(60/164), 40.8% (69/169) (χ²=0.93, P=0.334). The mean value of serum antibodies Absorbance value was 2.521±0.05 of 2015 and was 2.583±0.045 of 2016 in oyster culture area, the mean value was 2.249±0.05 in control group, there was a statistical difference among three group (F=13.28, P<0.001). The antibody prevalence in the three groups was 100%. BT50 geometric mean titer (GMT) of oyster culture area in 2015 was 423.1±40.11, culture group was 248.2±25.63, there was a statistical difference (t=3.73, P<0.001). Conclusion The population in oyster culture area does have more chance of exposure and infection GII.4 norovirus, Serum antibody of blocking ability in oyster culture areas is better than the general population in inland city. Suggesting that the population is more immunity resistant infected GII.4 norovirus.