OBJECTIVETo study viruses infecting Pinellia ternata in China.

METHODSymptom observation, DAS-ELISA and RT-PCR detection were applied.

RESULT AND CONCLUSIONDuring a survey in early spring, SMV and CMV were both commonly distributed as main viruses infecting P. ternata collected from different areas in China. But DsMV was the virus which infected P. ternate in natural condition. The infection ratio of cultivated P. ternate by SMV and CMV were 71.4% and 14.3% respectively for 21 samples collected from Ningbo, Zhejiang province; 100% and 44.4% for 18 samples from Xiaoshan, Zhejiang province; 61.9% and 33.3% for 21 samples from Hebei province; 50.0% and 41.7% for 12 samples from Anhui province; 16.7% and 16.7% for 12 samples from Sichuan province; 31.3% and none for 16 samples from Beijing. And the infection ratio of 25 wild samples from different areas of China infected by SMV and CMV were both 20.0%.

China ; Cluster Analysis ; Cucumovirus ; genetics ; isolation & purification ; DNA, Complementary ; chemistry ; genetics ; Mosaic Viruses ; classification ; genetics ; isolation & purification ; Pinellia ; virology ; Plant Diseases ; virology ; Plants, Medicinal ; virology ; Sequence Analysis, DNA

BACKGROUNDVascular access (VA) dysfunction is a major clinical complication in the hemodialysis population and has a direct effect on dialysis outcome. This study was conducted to explore the role of microinflammation in the VA dysfunction in maintenance hemodialysis patients.

METHODSForty-seven patients (male 35 and female 12) receiving maintenance hemodialysis were included for this study. They were divided into three groups: group 1 (n = 15), patients with initial hemodialysis and new arteriovenous fistula (AVF); group 2 (n = 18), patients treated with hemodialysis for long term with well-functional VA; group 3 (n = 14), maintenance hemodialysis patients with VA dysfunction. Biochemical parameters and serum tumor necrosis factor-alpha (TNF-alpha), interleukin 6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) were determined. High-sensitivity C-reactive protein (hs-CRP) was determined by latex-enhanced immuno-nephelometric method. Tissues of radial artery were taken from group 1 and group 3 for the histological study. Expression of CD68 and MCP-1 in the radial artery was determined by immunohistochemistry.

RESULTSSerum hs-CRP in group 3 was significantly higher than those in group 1 and group 2 ((7.40 +/- 2.42) mg/L vs (4.21 +/- 1.62) mg/L and (5.04 +/- 3.65) mg/L, P < 0.01 and P < 0.05, respectively). Serum TNF-alpha in group 3 was significantly higher than those in group 1 and group 2 ((64.03 +/- 9.29) pg/ml vs (54.69 +/- 12.39) pg/ml and (54.05 +/- 7.68) pg/ml, P < 0.05 and P < 0.01, respectively). Serum IL-6 in group 3 was also significantly higher than those in group 1 and group 2 ((70.09 +/- 14.53) pg/ml vs (56.43 +/- 10.11) pg/ml and (60.77 +/- 9.70) pg/ml, P < 0.01 and P < 0.05, respectively). Patients in group 3 had a thicker internal layer of vessels than in group 1 ((0.356 +/- 0.056) mm vs (0.111 +/- 0.021) mm, P < 0.01). Expression of CD68 and MCP-1 in the fistula vessel walls in group 3 were much higher than those in group 1 (P < 0.01). Moreover, serum hs-CRP level was positively correlated with the neointimal hyperplasia, the expression of CD68 and MCP-1 in fistula vessel (P < 0.01, respectively).

CONCLUSIONMicroinflammation might be involved in the dysfunction of AVF in patients with maintenance hemodialysis.

Adult ; Aged ; Arteriovenous Shunt, Surgical ; adverse effects ; C-Reactive Protein ; analysis ; Female ; Humans ; Immunohistochemistry ; Inflammation ; etiology ; Interleukin-6 ; blood ; Male ; Middle Aged ; Renal Dialysis ; adverse effects

Objective:To use polyamidoamine(PAMAM)dendrimer as gene delivery system for survivin gene anti- sense oligodeoxynucleotide(asODN)transfection for inhibition of HepG2 cancer cell growth.Methods:The first to the fifth generation of PAMAM and asODN were used to prepare a complex:PAMAM-asODN.The morphology of PAMAM- asODN was observed using agrose electrophoresis and atomic force microscope(AFM).PAMAM-asODN was then used to transfect HepG2 cells and cells transfected with asODN served as control.The transfection efficacy of PAMAM-asODN into HepG2 cells was observed under confocol microscope,the surviving mRNA expression was analyzed by RT-PCR,and the inhibition of HepG2 cell growth was determined by MTT assay.Results:Agrose electrophoresis showed strong complexing action between PAMAM and asODN and they formed a complex with a diameter of 25 nm.Confocol microscope showed the transfection efficacy of PAMAM-asODN was higher than that of asODN.RT-PCR showed a decreased expression of sur- vivin mRNA in PAMAM-asODN transfected cells.MTF results demonstrated that the growth of HepG2 cell was obviously inhibited after transfection of PAMAM-asODN and the inhibition rate increased with culture time,concentration of com- plex,the generation of PAMAM.PAMAM-asODN at 6.0?mol/L G4.0 resulted in a 55% inhibition of HepG2 cells 96 h after culture.Conclusion:PAMAM dendrimers can efficiently mediate the entry of survivin asODN into HepG2 cells,re- sulting in inhibition of HepG2 cells.PAMAM might be a promising gene carrier for potential molecular therapy of cancer.

By integration of traditional and Western medicine under particular historical conditions in China, some doctors of Western medicine are learning traditional Chinese medicine (TCM)and try to interconnect the two theories.To promote medical progress, they provide TCM services in clinic, such as acupuncture, herbal medicine, etc.But as we know that the laws are lacking for the doctors of Western medicine who want to carry out traditional Chinese medicine service.For the reason, the paper holds that at present there are three fundamental problems that should be solved.First of all, it must be clear that doctors should have qualifications and conditions to provide TCM service.Secondly, it must be clear that the doctors should have practicing scope to guarantee medical security and medical quality.Thirdly, it is imperative to strengthen TCM education and training for doctors.Now it is necessary to further strengthen the standardization of health supervision and law enforcement.

OBJECTIVETo evaluate the inhibitory effects of survivin antisense oligonucleotide (survivin-ASODN) mediated by polyamidoamine dendrimer (PAMAM) against the growth of subcutaneously transplanted colorectal cancer in nude mice.

METHODSNude mouse models bearing colorectal cancer was established by subcutaneous injection of SW620 cells. Survivin- OSADN (300 microg/L) was mixed with 4.06 microg/L PAMAM or liposome to prepare two transfection complexes, and their morphologies were observed by transmission electron microscope. The particle size of the prepared complexes was determined by laser particle size analyzer, and the zeta potential was measured. The encapsulation efficiency and the DNA release rate in vitro were determined by ultraviolet spectrophotometer. The transfection complexes were then directly injected into the xenografts of the tumor-bearing nude mice. The tumor volume changes were observed, and the expression of survivin in the transplanted tumor was measured by Western blotting.

RESULTSThe PAMAM-survivin-ASODN complex had a significantly smaller diameter and greater zeta potential than liposome-survivin-ASODN (P<0.01 and 0.05, respectively). The encapsulation efficiency was comparable between the two complexes. In in vitro condition, PAMAM-survivin-ASODN allowed sustained survivin-ASODN release for as long as 14 days, as compared with the 5 days for the liposome complex. After injection into the tumor xenografts, PAMAM-survivin- ASODN resulted in significantly lower expression of survivin protein in the transplanted tumors (P<0.05), and also in significantly greater reduction of the tumor volume than the liposome complex (P<0.05).

CONCLUSIONPAMAM can effectively deliver survivin-ASODN into transplanted colorectal tumor cells to reduce the expression of survivin and inhibit the tumor growth.

Animals ; Cell Proliferation ; drug effects ; Colorectal Neoplasms ; pathology ; Dendrimers ; Humans ; Inhibitor of Apoptosis Proteins ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microtubule-Associated Proteins ; genetics ; pharmacology ; Neoplasm Transplantation ; Oligonucleotides, Antisense ; pharmacology ; Polyamines ; pharmacology ; Repressor Proteins ; Tumor Cells, Cultured

The CrylA Crystal Protein from Bacillus thuringiensis is associated with DNA, but the role and sequences of these DNA molecules are unknown. CrylA bipyramidal crystals from B. thuringiensis strain 4.0718 was selectively dissolved and associated DNA was extracted from protoxin. The DNA was digested with Nde I to obtain 3 to 5 kb fragments and then the fragments were subcloned into pMD18-T vector, screening of recombinants were done by PCR-RFLP and sequencing. The ORF of cry1Ac gene was amplified by primers designed and then subcloned. The 3.5 kb BamH I and Sal I fragments of pMDX35 was inserted into the pET30a vector, giving 8.9 kb recombinant plasmid, pETX35. ETX35 strain were obtained by transformed pETX35 into B121 (DE3). A 141 kD fusion protein was superexpressed as inclusion bodies. Quantitative protein analysis indicated that the amount of 141 kD protein was above the level of 51.36% of total cellular protein. Plasmid pHTX42 constructed from shuttle vector pHT304 was transformed B. thuringiensis acrystalliferous strain XBU001 with electroporation to obtain the recombinant HTX42. The recombinant protein was found with a molecular mass of 130 kD on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Scanning analysis indicated that the expressed protein accounted up to 79.28% of total cellular proteins and accumulated in the cells mounted up to 64.13% of cellular dry weight. Under Atomic Force Microscopy (AFM), typical bipyramidal crystals from HTX42 strain were found with a size of 1.2 microm x 2.0 microm. Bioassay showed that these inclusion bodies of ETX35 strain and crystals from HTX42 strain were highly toxic against the larvae of Plutella xylostella. On such a base, constructing insecticidal recombinant and analyzing the source, structure, and function of the 20 kb DNA can be further achieved.
Animals ; Bacillus thuringiensis ; genetics ; Bacterial Proteins ; biosynthesis ; genetics ; pharmacology ; Cloning, Molecular ; Endotoxins ; biosynthesis ; genetics ; pharmacology ; Hemolysin Proteins ; biosynthesis ; genetics ; pharmacology ; Microscopy, Atomic Force ; Moths ; Plasmids ; Recombinant Proteins ; biosynthesis ; pharmacology

OBJECTIVETo study the influence of thermochemotheraphy on the expression of HSP70 in maxillofacial squamous cell carcinoma.

METHODS12 patients were treated with thermochemotheraphy twice a week, altogether 10 times. After 8 mg of Pingyangmycin infused, the patients were treated with microwave hyperthermia at 43 degrees C for 40 min. The part of carcinoma tissue was removed with surgical operations at before treatment and aftre five times of treatment. The expression of HSP70 in tumor cells was determined by SP immunohistochemcial method.

RESULTSThe expression of HSP70 in tumor cells was enhanced obviously by thermochemotherapy.

CONCLUSIONSpecial high expression of HSP70 in the tumor cells was induced by thermochemotherapy. With the antigen presenting action and other action, HSP70 have special antitumor effect.

Carcinoma, Squamous Cell ; HSP70 Heat-Shock Proteins ; Humans

OBJECTIVE: To explore the mechanism that how human enterovirus 71 (EV71) invades the brainstem and how intercellular adhesion molecules-1 (ICAM-1) participates by analyzing the expression and distribution of human EV71, and ICAM-1 in brainstem of infants with brain stem encephalitis. METHODS: Twenty-two brainstem of infants with brain stem encephalitis were collected as the experimental group and 10 brainstems of fatal congenital heart disease were selected as the control group. The sections with perivascular cuffings were selected to observe EV71-VP1 expression by immunohistochemistry method and ICAM-1 expression was detected for the sections with EV71-VP1 positive expression. The staining image analysis and statistics analysis were performed. The experiment and control groups were compared. RESULTS: (1) EV71-VP1 positive cells in the experimental group were mainly astrocytes in brainstem with [dark]-brown particles, and the control group was negative. (2) ICAM-1 positive cells showed [dark]-brown. The expression in inflammatory cells (around blood vessels of brain stem and in glial nodules) and gliocytes increased. The results showed statistical difference comparing with control group (P < 0.05). CONCLUSION The brainstem encephalitis can be used to diagnose fatal EV71 infection in infants. EV71 can invade the brainstem via hematogenous route. ICAM-1 may play an important role in the pathogenic process.
Astrocytes/pathology* ; Brain Stem/virology* ; Case-Control Studies ; Encephalitis, Viral/virology* ; Enterovirus A, Human/metabolism* ; Female ; Hand, Foot and Mouth Disease/virology* ; Humans ; Immunohistochemistry ; Infant ; Intercellular Adhesion Molecule-1/metabolism* ; Male