OBJECTIVETo study the influecnce of gentian leaf blight on the output and quality of rough gentian.

METHODThe same grade seedlings were transplanted, disease of every plant was investigated in autumn and the output of gentian was determined. HPLC was applied to determine the content of gentiopicroside and swertiamarin.

RESULTThe output decreased with the aggravation of the disease, and the decrease was obvious when the index of disease was above 60. The content of gentiopicroside and swertiamarin began to drop when the index of disease was above 70.

CONCLUSIONThe loss of output and the drop of quality are relatively heavy when the disease is serious. The loss of income is not obvious when the index of disease is under 60.

Gentiana ; chemistry ; growth & development ; Glucosides ; analysis ; Iridoid Glucosides ; Iridoids ; analysis ; Mitosporic Fungi ; growth & development ; Plant Diseases ; economics ; microbiology ; Plant Leaves ; microbiology ; Plants, Medicinal ; chemistry ; growth & development ; Pyrans ; analysis ; Pyrones ; analysis ; Quality Control

AIMUsing GeneChip to analyze the changes in genes expression of brain potassium, sodium and calcium channels after chronic treatment with nicotine.

METHODSAnimals were treated with nicotine at the doses of 2.4 mg/kg sc. twice a day for 14 days. RNA was extracted from the whole brain samples and converted to double-stranded cDNA and then to biotinylated cRNA. The biotinylated cRNA was fragmented, and hybridized to GeneChip (Affymetrix Rat Neurobiology U34). The chips were scanned with a probe array scanner, and the data were analyzed with the Affymetrix Microarray Analysis Suite (MAS). The GeneChip data were confirmed u sing RT-PCR.

RESULTSAfter treatment with chronic nicotine, transcripts of potassium, sodium and calcium channels showed altered expression. K+ channel: outward rectifier K+ channel and Ca2(+)-activated K+ channel were down-regulated, other voltage-dependent K+ channel including Kv2.3r were up-regulated. Voltage-dependent Na+ channel: beta2 subunit was increased, alpha subunit and beta1 subunit were decreased. Beta3 subunit of Ca2+ channel was up-regulated.

CONCLUSIONChronic exposure to nicotine not only desensitized nicotinic receptors, but also effected genes expression, of important ion channels, such as sodium channels, potassium channels and calcium channels.

Animals ; Brain ; drug effects ; metabolism ; Calcium Channels ; drug effects ; metabolism ; Gene Expression ; Male ; Nicotine ; pharmacology ; Potassium Channels ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sodium Channels ; drug effects ; metabolism

Chromogranin A ; metabolism ; Diagnosis, Differential ; Duodenal Neoplasms ; metabolism ; pathology ; surgery ; Ganglioneuroma ; metabolism ; pathology ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; Humans ; Male ; Middle Aged ; Neurofibroma ; metabolism ; pathology ; Paraganglioma ; metabolism ; pathology ; surgery ; Phosphopyruvate Hydratase ; metabolism ; S100 Proteins ; metabolism

OBJECTIVETo investigate the late results of using posterior restoration and three-column fixation to treat lumbar spondylolisthesis.

METHODSOne hundred and eighty-four patients with lumbar spondylolisthesis were collected from March 1999 to May 2007, they were treated by posterior restoration and fixation with single nail-grooved tail steel plate and fixed with cage (WDFC). Among these cases, 87 cases were fixed with one WDFC, 97 cases were used two WDFCs.

RESULTSAll patients were followed up for 8 to 69 months(averaged 23 months). According to Nakai standard, the results was excellent in 142 cases, good in 34, fair in 8, the excellent and good rates were 95.6%. Seventy-nine vertebraes with I degree spondylolisthesis were reduced after surgery. Eighty-seven vertebraes with II degree spondylolisthesis were reduced except 9 with I degree spondylolisthesis left. Twenty-one with III degree spondylolisthesis were reduced except 5 with I degree spondylolisthesis left; In 2 with IV degree spondylolisthesis, one with I degree spondylolisthesis left and the other with II degree spondylolisthesis left. The follow-up results showed that there was no statistical significance in the height of intervertebral space between preoperation and post-operation, and no recurrence was observed and no single nail-grooved tail steel plate and WDFC were loose or crashed.

CONCLUSIONPosterior restoration and three-column fixation is a positive modus operandi to treat lumbar spondylolisthesis,which can reduce excellently,keep the height of intervertebral space and stabilization of segment, obtain high rate of fusion, and cut down complication.

Adolescent ; Adult ; Aged ; Bone Plates ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Fusion ; instrumentation ; Spondylolisthesis ; surgery

OBJECTIVETo construct a recombinant lentiviral vector (pXZ208-BDDhFVIII) mediating B-domain-deleted human coagulation factor VIII (BDDhFVIII) gene and investigate its expression in HLF, Chang-Liver and MSC cells.

METHODSBDDhFVIII gene fragment was separated by endonuclease digestion and was cloned into the multiple cloning sites of pXZ208 to construct a recombinant lentiviral vector pXZ208-BDDhFVIII. Viral particles were prepared by means of three-plasmid cotransfection of 293T package cells by calcium phosphate precipitation. After infection, the coagulant activity of human FVIII in the culture medium of 293T, HLF, Chang-Liver and MSC cells was assayed by one-stage method. The gene transduction efficiency was assayed by flow cytometry (FCM). Furthermore, PCR was performed to test the integration of BDDhFVIII.

RESULTSThe infection rates of HLF, Chang-Liver and MSC were (74.52 +/- 7.57)%, (27.24 +/- 6.53)% and (42.34 +/- 5.84)% respectively. The activities of FVIII in supernatants of HLF, Chang-Liver and MSC were (54.1 +/- 5.6)%, (22.5 +/- 2.9)% and (12.5 +/- 2.7)% respectively. BDDhFVIII gene integration was detected in all the infected cells.

CONCLUSIONThe recombinant lentiviral vector pXZ208-BDDhFVIII was successfully constructed and efficiently integrated into target cells to express human FVIII activity in vitro.

Cell Line ; Factor VIII ; biosynthesis ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Plasmids ; Transfection

OBJECTIVETo investigate the influence of the prostaglandin E2 on the proliferation of the melanocytes in the full-thickness skin graft.

METHODSSixty-eight guinea-pigs were divided into experimental-1 group (skin graft), experimental-2 group (skin graft + diclofenac), and control groups. After the full-thickness skin graft, the dynamic changes of the prostaglandin E2 were measured and the proliferation of the melanocyte with its density was also evaluated by using histochemical and autoradiographic methods.

RESULTSIn the experimental-1 group, the content of PGE2 was increasing in seven days after the operation, continued to the one month, and then returned to the base level. The labelling indices of 3H-MC-TdR of the group was also increasing postoperatively between the second day and the fourteenth day, and reach a second peak after one month, then came to the normal level. The density of the melanocytes was decreasing rapidly 3 days after the surgery, then began to increase and exceeded over the normal level 21 days after the operation. However, in the experimental-2 group, the content of PGE2 decreased in two days after the surgery, and then showed the inclination similar to the experimental-1 group with the different points in narrower range. The number of melanocytes labelled by 3H-TdR began to increase at the first day after the surgery, which appeared earlier than the experimental-1 group and was similar in the changing tendency with a less extent. The density of MC showed the similar tendency to the experimental-1 group in a narrower changing range with both of increasing and decreasing. The density of the MC was much lower in 21 after the operation than the experimental-1 group and normal control group.

CONCLUSIONThe increased PGE2 in the earlier stage of the skin grafting could enhance the inflammatory reaction to the tissue, as well as the melanocytes. It may stimulate the proliferation of the MC with the result of increasing their density. The use of the diclofenac might reduce the inflammation and suppress the proliferation of melanocytes, and result in the skin with light color due to decreasing the number of MC in the epidermis of the graft.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Cell Count ; Cell Proliferation ; Diclofenac ; pharmacology ; Dinoprostone ; metabolism ; Epidermis ; Guinea Pigs ; Melanocytes ; cytology ; Skin ; Skin Transplantation ; Time Factors

OBJECTIVETo investigate the clinicopathological features of primary gastrointestinal non-Hodgkin's lymphomas (PGI-NHL) and their prognostic values.

METHODSThe clinical and pathological data of 216 patients diagnosed as PGI-NHL from Zhejiang Cancer Hospital were analyzed retrospectively. χ² test, log-liner model analysis, COX proportional hazard regression analysis and Life-table survival analysis were used to analyze the survival status of the patients by SAS 8.2 software, and Log-rank test was performed to couple the overall survival rates with different prognostic factors.

RESULTSTotally, the age of onset was 8 to 89 years with the median age of 56.5 years. Male versus female was 1.27∶1(121∶95). The most frequently involved location was stomach (147 cases, 68.1%), followed by ileocecus (25 cases, 11.6%), large intestine (20 cases, 9.3%), small intestine (17 cases, 7.9%) and multiple GI involvement (5 cases, 2.3%). 182 cases were classified as B cell lymphomas, 22 cases as T cell lymphomas, and 12 cases not classified exactly due to insufficient data. The 3-year and 5-year survival rates of the patients were 69.4% and 53.3%, respectively. Univariate analysis revealed that age＞60 years, ECOG≥2, high LDH level, stage Ⅲ-Ⅳ, IPI≥2, T cell type and intestinal involvement were predictors for poor prognosis. IPI≥2, T cell type and intestinal involvement were independent adverse predictors for prognosis by multiple COX proportional hazard regression analysis. Among different treatment groups, cases received chemotherapy combined with local radiotherapy gained the best survival status.

CONCLUSIONB-cell lymphoma was the main pathological type in PGI-NHL; IPI≥2, T-cell type and intestinal involvement are independent adverse predictors for prognosis; chemotherapy combined with local radiotherapy might be the choice of approach for advanced stage and aggressive PGI-HNL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Follow-Up Studies ; Gastrointestinal Neoplasms ; drug therapy ; pathology ; Humans ; Lymphoma, Non-Hodgkin ; drug therapy ; pathology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Young Adult

OBJECTIVETo investigate the expression and significance of Toll like receptor (TLR)4 mRNA and nuclear factor (NF)-kappaB p50 mRNA in human normal nasal mucosal cell before and after stimulation by LPS.

METHODSThe tissue was obtained from 15 normal middle turbinates (without rhinosinusitis). Every tissue was cultured in vitro, divided into 2 specimens. LPS was added into 15 specimens as LPS group and not added into other 15 specimens as control group. The pathomorphological characters of nasal mucosal cells were observed under optical microscope after stimulation by LPS. The expression of TLR4 mRNA and NF-kappaB p50 mRNA in normal human nasal mucosal cells were evaluated by in situ hybridization.

RESULTSNormal mucociliary agglutinated and mucosal cells were enlarged after stimulation by LPS; The expression of TLR4 mRNA in LPS group was higher than control group obviously. Their average density of light was 1.283 +/- 0.027 in LPS group while 0.538 +/- 0.038 in control group, and there was statistical significance between the two groups (t = 1.761, P < 0.05). The expression of NF-kappaB p50 mRNA was higher than control group obviously, and expressed in cellular nucleus predominantly. Their average density of light was 1. 668 +/- 0.037 in LPS group while 0. 372 +/- 0.052 in control group, and there was statistical significance between the two groups (t = 2. 624, P < 0. 01).

CONCLUSIONSLPS can activate the NF-kappaB p50 of human nasal mucosal cells through TLR4, and it may play some roles in stimulating and damage effect induced by LPS in nasal mucosal cells.

Adult ; Epithelial Cells ; metabolism ; Female ; Humans ; In Vitro Techniques ; Lipopolysaccharides ; Male ; Middle Aged ; NF-kappa B p50 Subunit ; metabolism ; Nasal Mucosa ; cytology ; metabolism ; RNA, Messenger ; genetics ; Toll-Like Receptor 4 ; genetics ; metabolism ; Young Adult

This study was aimed to study the potential effects of alloreactive NK cells (allo-NKs) in therapy of relapsed lung cancer after haploidentical hematopoietic stem cell transplantation using donor lymphocyte infusion (DLI). The F1 donors derived-NK cells were purified with MACS magnetic separation system, in which the proportion of the alloreactive Ly49A(+) cells was detected by flowcytometry and alloreactivity was measured by LDH method. The relapse model of lung cancer after haploidentical-HSCT was established. The distribution kinetic of infused donor lymphocytes in vivo was analyzed. The inhibition of relapse tumor, infiltration of lymphocytes in situ and fluctuation of 22 kinds of cytokines in serum after DLI were compared among different groups. The results showed that the infused donor cells of allo-NK group were accumulated mostly in lung, spleen and kidney for more than 48 hours with considerable higher levels according to the distribution kinetic curve. The sizes of relapse tumors between chemotherapy + PBS group and chemotherapy + DLI group showed no difference. However, the relapsed tumors in allo-NK + DLI group were significantly smaller than that in chemotherapy + DLI group or allo-NK + PBS group, in which increased infiltration of lymphocytes were defined in situ. The levels of cytokines such as MCP-1, IL-17, IL-12 and MCP-5 in serum of allo-NK + DLI group ascended compared with control group, though the level of IL-10 declined simultaneously. It is concluded that allo-NKs prolong the survival time of infused donor lymphocytes in vivo, promote the secretion of inflammatory cytokines and Th1-type of cytokines, and further improve the antitumor effects of DLI against relapse after transplantation.
Animals ; Cytokines ; blood ; Hematopoietic Stem Cell Transplantation ; methods ; Killer Cells, Natural ; cytology ; Lung Neoplasms ; therapy ; Lymphocyte Transfusion ; methods ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Neoplasm Recurrence, Local ; therapy ; Transplantation Conditioning ; methods

OBJECTIVETo study the therapy effect of adeno-associated viral gene transfer of sarcoplasmic reticulum Ca(2+)-ATPase 2a (SERCA2a) on chronic congestive heart failure (HF) in 30 days, and the possible mechanism of the therapy effect.

METHODSThe rats were divided into four groups: control group, HF group, Group HF + EGFP, and Group HF + SERCA2a. HF rats were obtained by creating descending aortic constriction. 0.9% sodium chloride solution, recombinant adeno-associated virus carrying enhanced green fluorescent protein gene (rAAV2.eGFP) and recombinant adeno-associated virus carrying SERCA2a gene (rAAV2.SERCA2a), were respectively delivered to pericardium of HF rats in different groups by intrapericardial injection with a trans-diaphragmatic approach. 30 days after gene transfer, hemodynamic parameters, SERCA2a protein expression and SERCA2a activity were analyzed. The proteome difference from rat hearts between Groups HF + SERCA2a and HF was detected by expression proteomics. Electrophoretic separation and quantitation of cardiac myosin heavy chain isoforms of hearts in different groups were performed at 30 days.

RESULTSAt 30 days, left ventricular function improved significantly in HF rats infected with rAAV2.SERCA2a (LVSP 146.52 +/- 13.86 vs 97.91 +/- 12.13, LVEDP 7.88 +/- 2.88 vs 21.15 +/- 3.57, LV +dp/dt 11 206.16 +/- 1730.11 vs 5948.93 +/- 1283.43, LV -dp/dt -8249.54 +/- 1076.09 vs -4497.50 +/- 652.12; P < 0.05). The recovered cardiac function in Group HF + SERCA2a rats was comparable to control rats, and had lower LV-weight/Body-weight ratio (2.46 +/- 0.17 vs 2.71 +/- 0.24, P < 0.05). Overexpression of SERCA2a increased both the protein content (0.39 +/- 0.11 vs 1.11 +/- 0.18, P < 0.05) and activity (228.62 +/- 25.11 vs 82.55 +/- 14.13, P < 0.05) up to nonfailing levels. Expressions of some energy metabolic enzymes in hearts of Group HF + SERCA2a were much higher than those of HF group. They included creatine kinase-muscle, enolase beta, fructose-bisphosphate aldolase, mitochondrial H(+)-ATP synthase alpha subunit, electron transfer flavoprotein alpha-subunit, H(+)-transporting ATP synthase and heart fatty acid binding protein. Downregulation of alpha-MHC and upregulation of beta-MHC in failing hearts were observed. Gene transfer of SERCA2a could increase the expression of alpha-MHC [(74.48 +/- 3.74)% vs (53.57 +/- 2.30)%, P < 0.05], and decrease the expression of beta-MHC [(25.52 +/- 3.74)% vs (46.43 +/- 2.30)%, P < 0.05] in HF rats. The expression profiles of alpha-MHC and beta-MHC and the ratio of alpha-MHC/beta-MHC were similar to those in controls.

CONCLUSIONSAdeno-associated viral gene transfer of SERCA2a can enhance SERCA2a functions, maintain calcium homeostasis, improve cardiac energy metabolism, and normalize the expression of cardiac myosin heavy chain isoforms in HF rats. As a result, the ventricular systolic and diastolic functions can be improved significantly, and the hypertrophy of the heart may be reduced in clinic. Adeno-associated viral gene transfer of SERCA2a demonstrated good therapy effects on HF rats.

Adenoviridae ; genetics ; Animals ; Calmodulin ; metabolism ; Disease Models, Animal ; Gene Transfer Techniques ; Genetic Therapy ; Heart Failure ; therapy ; Male ; Rats ; Rats, Sprague-Dawley ; Sarcoplasmic Reticulum ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; genetics