Cucumber mosaic virus was detected from infected Basella rubra L. with the indirect enzyme-linked immunosorbent assay. Total RNA was extracted from infected leaves and the cDNAs of coat protein gene of CMV-Ba were obtained by RT-PCR. The amplified cDNA fragments were then cloned into pMD 18-T vector and sequenced,the result showed that the CP gene was 657 nucleotides in length. This sequence was aligned with the obtained CP gene and some CMV strains or isolates of subgroup Ⅰ and subgroup Ⅱ in GenBank using DNA MAN software. The results showed that CMV-Ba shared 90.9%~93.8% and 76.1%~76.9% identity with the known CP genes of subgroup Ⅰ and Ⅱ respectively in nucleotide level,on the other hand,amino acids deduced from CMV-Ba CP gene shared 92.7%~97.7% and 72.4%~78.1% identity with the known CP protein of subgroup Ⅰ and Ⅱ,respectively. This suggested that CMV-Ba CP gene belongs to CMV subgroupⅠ.

AIMTo observe the effect of stretching left ventricles in the end of action potential on rabbit cardiac activity, and to investigate its possible mechanisms.

METHODSStretch (120 mmHg, 50 ms) was applied in the end of action potential by the pressure-clamp technique to observe if there would be any changes in rabbit cardiac activity and streptomycin (500 micromol/L) was used to identify the mechanism.

RESULTSStretch in the end of action potential caused arrhythmia (P < 0.05) and streptomycin blocked the above effect (P < 0.05).

CONCLUSIONStreptomycin could block the effect of stretching left ventricles in the end of action potential on rabbit cardiac activity, which indicates that stretch-activated ion channels involve it.

Action Potentials ; physiology ; Animals ; Arrhythmias, Cardiac ; etiology ; physiopathology ; Female ; Heart Ventricles ; physiopathology ; In Vitro Techniques ; Ion Channels ; physiology ; Male ; Mechanoreceptors ; drug effects ; Proprioception ; Rabbits ; Streptomycin ; pharmacology

Objective To investigated the effect of TangNaiKang (TNK) on VEGF protein expression of GK rats Thoracic aorta. Methods 51 male GK rats were divided into five groups randomly: model group, pioglitazone group, and TNK treatment group (low, immediate and high dose). Another 10 male Wistar rats were served as normal control group. GK rats were fed with high-grease forage, while normal control group was fed with a standard diet. Fasting blood glucose, general HE staining and VEGF protein expression were detected by immunohistochemistry. Results The fasting glucose had a significant decline in TNK treatment groups. HE staining showing TNK can ameliorate intima thickness, reduce hyperplasia of shallow vascular smooth muscle cell, and improve wavy and plexiform arrangement of elastic lamina. Immunohistochemistry also showed that TNK decreased VEGF protein expression of great vessels. Conclusion TangNaiKang can prevent and cure diabetic vascular complication of GK rats.

Objective To explore the relationship between intracellular calcium levels ([Ca2+]1) and reactive oxygen species (ROS) in sodium fluoride (NaF)-induced injury in human neuroblastoma SH-SY5Y cells. Methods The levels of [Ca2+]1 and ROS were measured in different exposed times(0,3,6,12,18,24 h) respectively after SH-SY5Y cells were exposed to 40 mg/L NaF in vitro, and the optimal expose time was selected. Furthermore, the changes of [Ca2+]1, ROS and LDH levels in 40 mg/L NaF-treated groups incubated with 38.23 mg/L BAPTA-AM or 380.40 mg/L ethylene glycol-bis-(beta-aminoethyl ether)-N, N, N', N'-tetraacetic acid (EGTA) or 16.32 mg/L N-acetyl-L-cysteine(NAC) also observed at the optimal expose time(12 h), respectively. Results At 3,6,12,18 and 24 h, [Ca2+]1 level(5620.0±226.3,4775.5±85.6,3312.3±87.5, 3047.0±75.0,2717.0±66.5) was significantly increased, and so was the ROS level(4449.53±324.61,7463.07±117.43,20 227.33±178.04,8817.56±200.13, 7975.61±92.90) except at 3 h, compared with 0 h(2115.0±24.0,4098.01±21.22, all P<0.05). The levels of [Ca2+]1 and ROS reached the peak at 3 h and 12 h, respectively. [Ca2+]1 and LDH levels in NaF-treated group [3279.5±94.0, (1057.50±64.35)U/L], NaF+NAC treated group[ 3583.0±350.7, (561.02±85.50)U/L], NaF+EGTA treated groups[3701.5±157.7, (1074.50±86.97)U/L], and BAPTA-AM treated group[2766.5±38.9, (521.43±40.80)U/L] had increased, compared with the control[2022.5±118.1, (186.97±8.73)U/L], the difference being statistically significant (P<0.05). ROS levels in NaF-treated group (19 003.04±332.34), and NaF+EGTA treated group(19 170.12±95.46) was higher than that in the controls(4060.98±145.66), the difference being statistically significant (P<0.05). NaF and NAC had antagonistic effect on ROS and LDH levels (F=976.11,43.54,P<0.05). And NaF and BAPTA-AM had antagonistic effect on [Ca2+]1, ROS and LDH levels (F=15.65,1515.53,115.00, P<0.05). Conclusions NaF-related calcium is released from the site of intracellular calcium storage, which induces ROS production, both of them caused cytotoxicity and the increase of LDH level in human neuroblastoma SH-SY5Y cells.

It is necessary to control the mechanical stimuli precisely in the studies of cardiac mechano-electrical feedback (MEF). In the present study a ventricular pressure-clamping system has been developed, which can be applied to isolated-perfused rabbit hearts. Controlled by a computer, this system not only can make the left ventricle follow a command defining the same pressure wave as that during a beating cycle under physiological condition, but also deliver mechanical stimuli with a proper waveform to the ventricle at a particular time phase. This system integrates multiple functions, including perfusing, pacing, recording of electrocardiogram and monophasic action potentials, and clamping and measuring of ventricular pressures in isolated-perfused hearts. Thus, it is a distinct system for investigating the phenomena and mechanisms of cardiac MEF at organ level.
Action Potentials ; Animals ; Constriction ; Electrocardiography ; Feedback ; Heart ; physiology ; In Vitro Techniques ; Rabbits ; Ventricular Pressure

OBJECTIVETo explore the relationship between two exonic polymorphisms of DNA repair gene XPC and the susceptibility to lung cancer.

METHODSGenotypes were determined by the primer introduced restriction analysis-PCR(PIRA-PCR) and the PCR-restriction fragment length polymorphism(PCR-RFLP) approaches, respectively, in 320 histologically-confirmed lung cancer cases and 322 age and sex frequency-matched cancer-free controls.

RESULTSMultivariate logistic regression analysis revealed that individuals carrying at least one 499Val variant allele (Ala/Val + Val/Val genotypes) had a significantly increased risk for lung cancer (adjusted OR=1.54; 95%CI: 1.11-2.14), compared with the wild-type genotype (499Ala/Ala). Furthermore, individuals with both putative risk genotypes had a significantly higher risk (adjusted OR=2.55; 95%CI: 1.45-4.52), compared with those with both wild-genotypes. In addition, a potential super multiplicative gene-environment interaction between Ala499Val genotypes and smoking on lung cancer risk was unveiled. The odds ratios of lung cancer for individuals with both putative risk genotypes were 2.63 (95%CI=1.23-5.62) in nonsmokers and 7.36 (95%CI=3.19-17.0) in smokers, respectively.

CONCLUSIONThese findings support the hypothesis that these two XPC variants may contribute to the risk of developing lung cancer.

Asian Continental Ancestry Group ; genetics ; China ; DNA-Binding Proteins ; genetics ; Exons ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Lung Neoplasms ; ethnology ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Risk Factors

OBJECTIVETo study the surgical reconstruction of secondary fracture deformities of the nasal-orbital ethmoid area.

METHODSTypical bicoronal and subciliary incisions or the adjacent scar incision were employed to expose the fractured area. The flattened nasal bone was narrowed by curved osteotomy along the medial orbital rims and trimed with a bur. 2-3 pieces of cranial outer table were used to reconstruct the nasal framework, which were fixed to the frontal bone with mini-plates. After the medial orbital wall and orbital floor were exposed, the herniated orbital contents were released and reduced to the orbital cavity. The fractured orbital walls were repaired precisely with autogenous cranial outer table or Medpor. The medial canthal tendons were anchored superior-posteriorly to the lacrimal fossa with transnasal wires.

RESULTSFrom December 1996 to December 2001, 34 cases of severe nasal-orbital ethmoid fracture deformities were repaired with this technique. Of them, 12 cases had combined orbital-zygomatic fracture, 4 cases had fontal sinus fracture, 1 case had Le Fort II and 1 case had Le Fort III fracture. All the patients recovered well and their facial appearance improved greatly.

CONCLUSIONSThe key points for surgical reconstruction of the periorbital post-fracture deformities are narrowing the flattened nasal root by curved osteotomy, the nasal framework reconstruction with autogenous bone graft, the orbital wall repair to correct enophthalmos, and most importantly, the medial canthal tendon reduction and canthal plasty.

Adolescent ; Bone Transplantation ; methods ; Cicatrix ; surgery ; Enophthalmos ; surgery ; Ethmoid Bone ; injuries ; Humans ; Nasal Bone ; injuries ; Nose Deformities, Acquired ; surgery ; Orbit ; surgery ; Orbital Fractures ; surgery ; Osteotomy ; methods ; Paranasal Sinuses ; injuries ; surgery ; Polyethylenes ; Reconstructive Surgical Procedures ; Skull Fractures ; etiology ; surgery ; Tendons ; surgery

In this study, 17 kinds of Dendrobium species of Fengdous including 39 individuals were collected from 4 provinces. Mitochondrial gene sequences co I, nad 5, nad 1-intron 2 and chloroplast gene sequences rbcL, matK amd psbA-trnH were amplified from these materials, as well as nrDNA ITS. Furthermore, suitable sequences for identification of Dendrobium species of Fengdous were screened by K-2-P and P-distance. The results showed that during the mentioned 7 sequences, nrDNA ITS, nad 1-intron 2 and psbA-trnH which had a high degree of variability could be used to identify Dendrobium species of Fengdous. However, single fragment could not be used to distinguish D. moniliforme and D. huoshanense. Moreover, compared to other combined fragments, new type combined fragments nrDNA ITS+nad 1-intron 2 was more effective in identifying the original plants of Dendrobium species and could be used to identify D. huoshanense and D. moniliforme. Besides, according to the UPGMA tree constructed with nrDNA ITS+nad 1-intron 2, 3 inspected Dendrobium plants were identified as D. huoshanense, D. moniliforme and D. officinale, respectively. This study identified Dendrobium species of Fengdous by combined fragments nrDNA ITS+nad 1-intron 2 for the first time, which provided a more effective basis for identification of Dendrobium species. And this study will be helpful for regulating the market of Fengdous.
DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Dendrobium ; classification ; genetics ; Genes, Chloroplast ; Genes, Plant ; Introns ; Plants, Medicinal ; classification ; genetics

OBJECTIVETo study the effect of Huoxuezhitong recipe on the pathway of mitochondrial apoptosis and the expression of related gene (Caspase-3, -9, Bax, Bcl-2) in epidural scar tissue.

METHODSA total of 60 New Zealand rabbits, ranging in weight from 2.5 to 3.0 kg, were randomly divided into sham operative group (group A), control group (group B), sodium hyaluronate group (group C), Huoxuezhitong recipe group (group D, included Danggui 20 g, Chishao 20 g, Honghua 20 g, Ruxiang 15 g, Myrrh 15 g etc.) with 15 rabbits in each group. Except for group A, L4,5 vertebral plate were resected leading to 1.0 cm x 1.0 cm dura mater exposed area, which were covered with 0.5 ml sodium hyaluronate in group C, covered with same amount of saline in group B and D. At 2 weeks after operation, the rabbits of group D were administered with Huoxuezhitong recipe (2.5 ml/kg, once a day, continuous 14 d). Five rabbits of each group were selected randomly and were killed at 2, 4 and 8 weeks after operation; and the expression of Caspase-3, -9, Bax, Bcl-2 in scar tissue were surveyed by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe results by RT-PCR showed significant different in expressions of Caspase-3, -9, Bax, Bcl-2 of group A and others groups at each period (P < 0.01). At 2, 4, 8 weeks after operation, compared with group B, the expressions of Caspase-3, -9, Bcl-2 in group C and D were decreased (P < 0.05), but the expression of Bax in group C and D were increased (P < 0.05). At 8 weeks after operation, the expressions of Caspase-3, -9, Bax, Bcl-2 in group D was significant decreased compared with group C (P < 0.05).

CONCLUSIONHuoxuezhitong recipe can regulate the levels of expression of Caspase-3, -9, Bax, Bcl-2 mRNA and further induce the pathway of mitochondrial apoptosis, thereby, achieve the effect of prophylaxis for the proliferation and conglutination of epidural scar tissue.

Actins ; genetics ; metabolism ; Animals ; Apoptosis ; drug effects ; genetics ; Carthamus tinctorius ; Caspase 3 ; genetics ; metabolism ; Caspase 9 ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Genes, bcl-2 ; drug effects ; genetics ; Mitochondria ; drug effects ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; bcl-2-Associated X Protein ; genetics ; metabolism

Bletilla striata has been used as traditional Chinese medicine for several centuries. In recent years, the quality and quantity of wild B. striata plants have declined sharply due to habitat deterioration and human over-exploitation. Therefore, it is of great urgency to evaluate and protect B. striata wild plant resource. In this study, sequence-related amplified polymorphism (SRAP) markers were applied to assess the level and pattern of genetic diversity in twelve populations of B. striata. The results showed a high level of genetic diversity (PPB = 90.48%, H = 0.349 4, I = 0.509 6) and moderate genetic differentiation among populations (G(st) = 0.260 9). Based on the unweighted pair-group method with arithmetic average (UPGMA), twelve populations gathered in three clusters. The cluster 1 included four populations. There are Nanjing, Zhenjiang, Xuancheng and Hangzhou. The seven populations which come from Hubei Province, Hunan Province, Jiangxi Province and Guizhou Province belonged to the cluster 2. The cluster 3 only contained Wenshan population. Moreover, Mantel test revealed significant positive correlation between genetic distances and geographic distances (r = 0.632 9; P < 0.000 1). According to the results, we proposed a series of conservation consideration for B. striata.
China ; Genetic Markers ; Genetic Variation ; Genetics, Population ; Orchidaceae ; genetics ; Phylogeny ; Plants, Medicinal ; genetics