AIM: To provide experimental evidence for gene therapy of thrombophilia disease, we constructed the eukaryotic expression plasmid with human thrombomodulin (hTM) gene and observed the alteration of hTM expression on the surface of human umbilical vein endothelial cells (HUVECs) with and without the reconstructive plasmid. METHODS: The whole expressive fragment of hTM gene was amplified by PCR from human genome. Both hTM gene and pcDNA3.1(+)/neo empty vector was digested by HindⅢ and EcoRⅠ. Two digested fragments were ligated into pcDNA3.1/hTM with T_4DNA ligase. After identifying, the reconstructive plasmid transfected into HUVECs using lipofectin. The hTM antigen on the HUVECs was detected by immunohistochemistry. RESULTS: The hTM reconstructive plasmid was confirmed by double endonuclease redigesting and sequencing. About 10% HUVECs were transfected by pcDNA3.1/hTM plasmid with lipofectin and the high-level hTM was detected on the transfected cells. CONCLUSION: We constructed the pcDNA3.1/hTM plasmid successfully, and it could be expressed on the HUVECs. [

Animals ; Bites and Stings ; virology ; Cats ; Child ; Epstein-Barr Virus Infections ; diagnosis ; Herpesvirus 4, Human ; Humans ; Lymphadenitis ; diagnosis ; virology ; Male

To explore the new method of discriminating Cistanche deserticola, Cynomorium songaricum and Orobanche pycnostachya by using PCR amplification of specific alleles. 30 samples of the different C. deserticola, 21 samples of C. songaricum and O. pycnostachya were collected. The total DNA of the samples were extracted, the ITS sequences from C. deserticola, C. songaricum and O. pycnostachya were amplified by PCR and sequenced unidirectionally. These sequences were aligned by using ClustulW. Specific primer was designed according to the ITS sequences of specific alleles, and PCR reaction system was optimized. Additionally, compare with the identification of specific PCR method and DNA sequence analysis method. The result showed that the 331 bp identification band for C. deserticola and the adulterants not amplified bands by a single PCR reaction, which showed good identification ability to the three species. PCR amplification of specific alleles can be used to identify C. deserticola, C. songaricum and O. pycnostachya successfully.
Alleles ; Cistanche ; classification ; genetics ; DNA Primers ; genetics ; DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Drug Contamination ; prevention & control ; Phylogeny ; Polymerase Chain Reaction ; methods

. Conclusions Two multiplex RT-PCR assays show high consistency with common RT-PCR. The multiplex RT-PCR assays were initially established.

In this study, the rat type 2 diabetes mellitus (T2DM) model was established through tail vein injection with low dose of streptozotocin (STZ) and high fat diet for 8 weeks, and then treated with Jiaotai Pill. The oral glucose tolerance test (OGTT), fasting serum insulin (FINS), free fatty acid(FFA) levels and blood lipid were assayed. HOMA-IR was calculated. Pancreatic pathology was performed. And pancreatic triglyceride (TG) content was examined by the lipid extraction method. Pancreatic islet cell apoptosis were detected by terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL). According to the results, the model group showed abnormal OGTT, increased FINS, HOMA-IR, FFA, lipid disorder, obvious fat accumulation and significantly increased TG content in pancreatic tissues, and enhanced pancreatic islet cell apoptosis. Compared with the model group, the Jiaotai Pill group displayed improved OGTT, reduced FINS, HOMA-IR, FFA, recovered lipid disorder, decreased fat accumulation and significantly declined TG content in pancreatic tissues, and lowered pancreatic islet cell apoptosis. In summary, Jiaotai pill could effectively treat type 2 diabetes in rats. Its mechanism may be related to the reduction in pancreatic fat accumulation and islet cell apoptosis.
Animals ; Apoptosis ; drug effects ; Diabetes Mellitus, Type 2 ; drug therapy ; metabolism ; physiopathology ; Drugs, Chinese Herbal ; administration & dosage ; Fats ; metabolism ; Glucose Tolerance Test ; Humans ; Islets of Langerhans ; cytology ; drug effects ; Male ; Pancreas ; drug effects ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo explore the relationship between Chinese medicine syndrome patterns (CMSP) and their associated factors in patients with type 2 diabetic nephropathy stage III (DN2-3).

METHODSRetrospective analysis was conducted on 209 patients with type 2 diabetes mellitus (T2DM). The patients were allocated into two groups, the DN2-3 group and the control group. Some related clinical materials and laboratory indexes, including age, course of disease, body mass index (BMI), glycosylated hemoglobin A1c (HbA1c), systolic blood pressure (SBP), diastolic blood pressure (DBP), blood levels of total cholesterol (TC), triglyceride (TG), high and low density lipoprotein (HDL and LDL), serum creatinine (Scr) and microalbuminuria (MALB) as well as their CMSP (both the essential syndrome and the superficial syndrome) in the DN2-3 group were collected and compared.

RESULTSSignificant differences were found between the DN2-3 group and the control group in aspects of course of disease (months, 107.74 +/- 96.19 vs. 82.03 +/- 79.10), BMI (kg/m2, 26.25 +/- 4.02 vs. 24.95 +/- 3.56) and Scr level (mmoL/L, 71.93 +/- 24.24 vs. 65.91 +/- 13.70, P < 0.05). The qi-yin deficiency SP (38 cases, 36.19%), and the blood stasis (51 cases, 48.58%) presented as the dominant essential and superficial CM-SP respectively in DN2-3 patients, holding the highest proportion. Analysis on the relationship of associated indices among patients with different CMSP showed statistical differences presented in level of MALB, i.e. which in pi-shen qi-deficiency SP (128.77 +/- 103.59 mg/24 h) was higher than in yin-deficiency dryness-fire SP and qi-yin deficiency SP (88.43 +/- 68.93 mg/24h and 82.60 +/- 55.22 mg/24 h, P < 0.05); it also presented in HbA1c (%) and TG levels(mmol/L), those in stasis SP were 10.73 +/- 2.71 and 2.29 +/- 1.58 ), in dampness SP were 8.80 +/- 2.19% and 4.37 +/- 5.92, and in stasis-phlegm SP were 8.83 +/- 2.09 and 2.40 +/- 2.18 (all P < 0.05).

CONCLUSIONSThe risk factors for occurrence of DN2-3 may be the course of disease, BMI and Scr. Qi-yin deficiency with blood-stasis is the most commonly encountered syndrome in patients with DN2-3. Relations of MALB with Pi-Shen qi-deficiency pattern; HbA1c with blood-stasis pattern, and TG with dampness syndrome are distinctly exhibited in them.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Albuminuria ; physiopathology ; Diabetes Mellitus, Type 2 ; physiopathology ; Diabetic Nephropathies ; physiopathology ; Diagnosis, Differential ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Retrospective Studies ; Young Adult

OBJECTIVETo study the early change of serum nitric oxide (NO) after acute heat exposure with trauma and the effect of NO on mean arterial pressure (MAP), thus to provide theoretical basis for studying the mechanism of NO effect in acute stress.

METHODSThe rabbit model of acute heat exposure combined with trauma was established. The animals were divided into four groups, including control, trauma, hyperthermia and hyperthermia combined with trauma. The levels of NO were measured at different time points: 0 h, 1 h, 2 h and MAP was monitored throughout the whole experiment.

RESULTSThe concentration of NO declined at first and then increased at 1 h or so after acute heat exposure and trauma. The levels of NO in hyperthermia with trauma group at 1 h, 2 h were (42.75 +/- 8.24), (59.54 +/- 9.05) micro mol/L respectively (P < 0.05), while those in control group were (56.63 +/- 3.79) and (55.22 +/- 7.15) micro mol/L, the difference at 1h between two groups was significant (P < 0.05). Under the circumstance of hyperthermia and trauma, the level of MAP declined to the lowest point at 60 - 70 min and then showed a transient rise, after that, the level declined rapidly.

CONCLUSIONSAt the early stage of acute heat exposure and trauma, the concentration of serum NO declined at first and then increased, and had certain relationship with the change of MAP.

Animals ; Blood Pressure ; Cytokines ; biosynthesis ; Hot Temperature ; Male ; Nitric Oxide ; blood ; Rabbits ; Wounds and Injuries ; blood ; physiopathology

OBJECTIVETo transfect pcDNA3.1/hTM plasmids containing human thrombomodulin (hTM) gene into human umbilical vein endothelial cells (HUVECs), and investigate the expression of hTM and anticoagulating function of transfected HUVECs.

METHODSHUVECs were transfected with pcDNA3.1/hTM by lipofectin. Expression of hTM mRNA was determined by semi-quantitative RT-PCR, hTM antigen on HUVECs membrane by immunohistochemistry, and activated protein C (PC) in HUVECs by chronometry. By using a semiautomatic coagulator, the effect of the reacting liquid from transfected HUVECs mixed with PC from normal peripheral blood was assayed.

RESULTSAbout 10% HUVECs were transfected by pcDNA3.1/hTM with high-level hTM mRNA and protein expression. Activated PC produced by pcDNA3.1/hTM group, pcDNA3.1(+)/neo group and untransfected group was (2.80 +/- 0.43) microg/ml, (0.75 +/- 0.08) microg/ml and (0.85 +/- 0.11) microg/ml, respectively. APTT was (51.68 +/- 2.73) s, (38.38 +/- 2.44) s, (39.65 +/- 2.39) s, (33.93 +/- 1.73) s and (34.60 +/- 1.86) s and PT was (21.89 +/- 1.66) s, (20.56 +/- 1.74) s, (20.42 +/- 2.04) s, (19.57 +/- 1.36) s and (20.16 +/- 1.35) s in pcDNA3.1/hTM group, pcDNA3.1(+)/neo group, untransfected group, inactivating PC group and control, respectively.

CONCLUSIONSThe pcDNA3.1/hTM plasmid could be transfected into endothelial cells and expressed biologically functioning hTM protein on HUVECs membrane. Activated PC could inhibit intrinsic coagulation pathway obviously with slight effect on extrinsic pathway.

Cells, Cultured ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; cytology ; Gene Expression Regulation ; Genes, Transgenic, Suicide ; Humans ; In Vitro Techniques ; Partial Thromboplastin Time ; Plasmids ; Protein C ; Prothrombin Time ; Thrombomodulin ; genetics ; Transfection ; Umbilical Veins ; cytology

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Leukemia, Prolymphocytic, B-Cell ; diagnosis ; Male ; Middle Aged ; Retrospective Studies

Mongolian folk medicine, the important part of Mongolian medicine, is the main means, method and weapon of disease prevention, treatment and health care. Mongolian materia medicas are the important literatures of guiding the healthy development of the modern Mongolian medicine with a long and dazzling history. Since the founding of new China, a new history chapter of Mongolian folk medicine was opened under the attention and support from all levels of party and government. This paper intends to provide comprehensive insight into the rapid development of Mongolian folk medicine. The resources, phytochemistry, quality standard, pharmacology, dosage forms reform and production were reviewed to expound the process that Mongolian folk medicine was developed from traditional practices to scientific development
Medicine, Mongolian Traditional ; standards ; Science