AIM: To construct eukaryotic expression vector of human vascular endothelial growth factor (VEGF) gene, and to explore the effects of VEGF gene on proliferation and chemosensitivity of leukemia cells. METHODS: The recombinant eukaryotic express plasmid pEGFP-C1-hVEGF_(165) was constructed with conventional gene engineering methods. The pEGFP-C1-hVEGF_165 was transfected into leukemia cell K562. The proliferation of the recombinant eukaryotic expression plasmid was analyzed by CCK8 method. The level of VEGF mRNA was detected by reverse transcription and quantitative polymerase chain reaction (RT-PCR). The expression level of VEGF protein was determined by ELISA in cell culture medium and cells. The cell cycle was detected by flow cytometry. RESULTS: VEGF_(165) eukaryotic expression vector was successfully constructed, confirmed by enzyme digestion and sequencing. Compared with the K562 cells transfected pEGFP-C1, the K562 cells transfected with pEGFP-C1-hVEGF_(165) grew faster and expressions of VEGF mRNA and protein increased significantly. In addition, K562 cells transfected with pEGFP-C1-hVEGF_(165) had relatively higher cell survival rate to chemotherapy drugs homoharringtonine(HHT) and fluorouracil(FU). CONCLUSION: VEGF_(165) eukaryotic expression vector increases the level of VEGF mRNA and protein expression, accordingly promotes the proliferation of leukemia cells and decreases the sensitivity of leukemia cells to HHT and FU.

Apolipoprotein A5(ApoA5)level and other indices were determined in patients with type 2 diabetes mellitus and healthy individuals.Compared to control group,ApoA5 level in the diabetic group was lower (P

DEC1 is a basic helix-loop-helix(bHLH)transcription factor involved in the regulation of cell prolif-eration;cell differentiation;lymphocytes maturation;circadian rhythms;immune response and response to hypoxia.Recent studies have revealed that the expression of DEC1 is abnormally high in various tumor tissues and cells.In addition;the expression of DEC1 in the tumor tissue is related to the malignancy of various cancer types.This paper;therefore;focuses on the relationship between DEC1 and proliferation;apoptosis;cellular senes-cence;invasion and metastasis of tumor cells;in order to elaborate the role of DEC1 in the pathogenesis and pro-gression of tumor.

Objective To study the clinical features of paroxysmal sympathetic hyperactivity (PSH).Methods The clinical data,imaging and electroencephalography (EEG) of 10 patients with PSH was analyzed retrospectively.Results Of the 10 patients with PSH,9 were males and 1 was a female.The overall age of all the patients was (37.6 ± 19.1) years,ranging from 15 to 78 years.The primary diseases included traumatic brain injury 5 cases,intracranial hemorrhage 1 case,cerebral infarction 1 case,hypoxic ischemic encephalopathy 1 case,arachnoid cyst 1 case and cryptococcal meningoencephalitis 1 case.All patients developed at least 5 of 7 features which contained paroxysmal agitation,hyperthemia,diaphoresis,tachypnea,tachycardia,hypertension and dystonia.PSH occurred within 24 hours after brain injury in 3 patients; 24 hours to 3 weeks in 5 patients ; 5 months in 1 patient; 9 years in 1 patient.The frequency varied from one time in several days to several times in one day.The duration varied from 1 minute to 3 hours.The episodes in 4 patients occurred more often at night,1 around palinesthesia and the frequency of other 5 patients showed no differences between day and night.There were 2 cases appeared sober-minded,and the states of consciousness of the other 8 cases were hard to judge during PSH.The Glasgow Coma Scale scores of 8 cases were 3 to 8 points and the other 2 cases were 15 points.No epileptic-form activity was detected by EEG and no particular lesions were responsible.Neuro-imaging examinations suggested frontal lobe,temporal lobe,parietal lobe,occipital lobe,basal ganglion,pons and lateral ventricle were damaged.And 9 patients received an ineffective antiepileptic drug treatment.The efficacy in the management of PSH with dopamine agonists combining with β-blockers was observed.Two patients achieved complete remission,6 patients had a reduction in episode frequency,1 patient showed no response to the therapy and 1 patient discharged and could not be connected.Conclusions PSH can occur after various types and different degrees of brain injury.PSH is often misdiagnosed as epilepsy,and anticonvulsant therapies are useless.PSH receives good responses to β-blockers and dopamine agonists.

Cadherins ; metabolism ; Gastric Mucosa ; metabolism ; Humans ; Integrin beta1 ; metabolism ; Kangai-1 Protein ; metabolism ; Lymphatic Metastasis ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Staging ; Stomach Neoplasms ; metabolism ; pathology

Objective:To observe the effect of moxibustion on T lymphocyte subsets in peripheral blood of rats with gastric cancer. Methods:Sixty healthy Sprague-Dawley (SD) rats were adaptively fed for one week. By the random number table method, 10 rats were randomly selected as a blank group, and 12 rats were randomly selected to simulate the tumor transplantation process; after transplantation, 10 rats were randomly selected as a sham operation group. The remaining 38 rats were used to prepare gastric cancer models by gastric transplantation of the Walker-256 tumor tissue; 8 rats were randomly selected to verify the successful modeling after 7 d; the remaining 30 rats were randomly divided into a model group, a moxibustion group and an infrared group by the random number table method, with 10 rats in each group. From the first day of enrollment, the rats in the moxibustion group received mild moxibustion at Zhongwan (CV 12), Guanyuan (CV 4) and bilateral Zusanli (ST 36) (the first group) and bilateral Pishu (BL 20) and Weishu (BL 21) (the second group), and the two groups of acupoints were alternated every other day. The rats in the infrared group received infrared radiation on the stomach area and the area on the back between the T12-T13 spinous processes, the two areas were alternated every other day. Rats in the moxibustion group and the infrared group were treated for 20 min each time, once a day for 21 d. Rats in the blank group, the sham operation group, and the model group were simultaneously grasped and fixed, and no other treatment was performed. After 21 d of intervention, the rats in each group were fasted for 12 h, and blood was collected from the orbits. The numbers of CD3+, CD3+CD4+, CD3+CD8+ T lymphocytes in peripheral blood were determined by flow cytometry, and the ratio of CD3+CD4+/CD3+CD8+ was calculated. The rats were sacrificed and the thymus was dissected under sterile conditions to calculate the thymus index. Results:Compared with the blank group, the thymus index, peripheral blood CD3+, CD3+CD4+, CD3+CD8+ and CD3+CD4+/CD3+CD8+ ratio in the sham operation group did not change significantly (allP>0.05). Compared with the blank group, the thymus index of the model group was increased (P<0.05), the CD3+ and CD3+CD8+ T lymphocytes were increased (bothP<0.01), and the CD3+CD4+/CD3+CD8+ ratio was decreased (P<0.05). Compared with the model group, the thymus index of the moxibustion group was increased (P<0.01), and CD3+, CD3+CD4+ and CD3+CD8+ T lymphocytes and the ratio of CD3+CD4+/CD3+CD8+ in peripheral blood were increased (allP<0.05). Compared with the infrared group, the thymus index of the moxibustion group was significantly increased (P<0.05), the CD3+ and CD3+CD4+ T lymphocytes in the peripheral blood were significantly increased (bothP<0.01), and the CD3+CD8+ was increased (P<0.05). Conclusion:Moxibustion can significantly increase the thymus index of gastric cancer-bearing rats and activate CD3+CD4+ and CD3+CD8+ T lymphocytes in peripheral blood.

Blood Gas Analysis ; Continuous Positive Airway Pressure ; instrumentation ; Humans ; Infant ; Infant, Newborn ; Pneumonia ; blood ; physiopathology ; therapy ; Respiration

OBJECTIVEIsobaric tags for relative and absolute quantitation (iTRAQ) combined with mass spectrometry were used to screen differentially expressed plasma proteins in cold stress rats.

METHODSThirty health SPF Wistar rats were randomly divided into cold stress group A and control group B, then A and B were randomly divided into 3 groups (n = 5): A1, A2, A3 and B1, B2, B3. The temperature of room raising was (24.0 +/- 0.1) degrees C, and the cold stress temperature was (4.0 +/- 0.1) degrees C. The rats were treated with different temperatures until 12 h. The abdominal aortic blood was collected with heparin anticoagulation suction tube. Then, the plasma was separated for protein extraction, quantitative, enzymolysis, iTHAQ labeling, scx fractionation and mass spectrometry analysis.

RESULTSTotally, 1085 proteins were identified in the test, 39 differentially expressed proteins were screened, including 29 up-regulated proteins and 10 down-regulated proteins. Three important differentially expressed proteins related to cold stress were screened by bioinfonnatics analysis (Minor histocompatihility protein HA-1, Has-related protein Rap-1b, Integrin beta-1).

CONCLUSIONIn the experiment, the differentially expressed plasma proteins were successfully screened in cold stress rats. iTRAQ technology provided a good platform to screen protein diaguostic markers on cold stress rats, and laid a good foundation for further. study on animal cold stress mechanism.

Animals ; Blood Proteins ; chemistry ; Cold Temperature ; Mass Spectrometry ; Rats ; Rats, Wistar ; Stress, Physiological

OBJECTIVEIt could give some theory support of confirming the secondary metabolism organ and regulation of echinacoside in Cistanche tubulosa by searching parasitic growth of C. tubulosa ahd echinacoside variation in different organs of host and parasite.

METHODThe echinacoside content was analyzed by HPLC. The relationship between dry matter accumulation and echinacoside accumulation of C. tubulosa as the well as root diameter of host were comparatively analyzed.

RESULTWith the increase of dry matter accumulation of C. tubulosa, echinacoside accumulation increased significantly, and both of them were in significantly positive correlated with the root diameter of host. Echinacoside content in haustorium phloem was 15.53%, higher than that of haustorium xylem, C. tubulosa plant and other organs.

CONCLUSIONHaustorium phloem was probably the secondary metabolism organ of echinacoside in C. tubulosa.

Cistanche ; growth & development ; metabolism ; physiology ; Glycosides ; metabolism ; Host-Parasite Interactions ; Plant Leaves ; metabolism ; Plant Roots ; anatomy & histology ; metabolism ; Plant Stems ; metabolism ; Tamaricaceae ; anatomy & histology ; metabolism ; parasitology

The contents of adenosine, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxybenzaldehyde, parishin and sulfur dioxide residue were compared in differently-processed Gastrodiae Rhizoma to provide the basis for a reasonable processing method of Gastrodiae Rhizoma. The analysis was performed on a Merck Purospher STAR column (4.6 mm x 250 mm, 5 μm) with a mobile phase consisting of methanol and water (containing 0.1% formic acid) under gradient elution at a flow rate of 1.0 mL x min(-1). The eluates were detected at 270 nm, and the column temperature was 35°C. The content of adenosin, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxy-benzaldehyde and parishin in processing of boiling or sulfur-fumigated were lower than that of in processing of steaming. Furthermore, the sulfur dioxide residue of sulphur-fumigated groups exceed 400 mg x kg(-1). This stable and reliable method will contribute to the quality control of different processed Gastrodiae Rhizoma.
Drug Contamination ; Drugs, Chinese Herbal ; chemistry ; Gastrodia ; chemistry ; Sulfur Dioxide ; analysis ; Technology, Pharmaceutical ; methods