Objective: To further understand the role of folic acid supplements rivaling MTHFR gene silencing in pathogenesis of NCLP, RNA interference (RNAi) was applied to knock down MTHFR in mouse embryonic palatal mesenchymal (EPM) cells. Methods: MTHFR ShRNA expression vector were transfected into the primary cultured EPM cells. MTT was used to observe cell proliferation after MTHFR gene silencing. FCM was used to observe cell cycle after MTHFR gene silencing. Results: The results showed the cells proliferation had an inequality amelioration after using folic acid supplements in MEPM cells with MTHFR gene silencing. Using folic acid supplements rivaled the effect of MTHFR gene silencing had a dose-dependent manner. Using 20 μg/ml folic acid supplements could improve the cell proliferation to achieve normal level of cell proliferation. Conclusion: MTHFR gene is an important candidate gene of NCL/P. Using folic acid supplements could prevent teratogenic MTHFR gene silencing for embryonic palate development.

Environmental Pollution ; prevention & control ; Health Education ; Health Knowledge, Attitudes, Practice ; Humans ; Iatrogenic Disease ; prevention & control ; Medical Laboratory Personnel ; psychology

OBJECTIVETo explore the effect of drug-containing serum of Chinese herbal compounds [Xiongshao Capsule (XS, for activating blood) and Huanglian Capsule (HL, for dispelling toxin)] on tumor necrosis factor-alpha (TNF-alpha)-induced adherence between human umbilical vein endothelial cells (HUVECs) and polymorphonuclear neutrophils (PMN), inflammatory reaction and expression of related proteins in mitogen-activated protein kinase (MAPK) pathway.

METHODSThirty-two rats were randomly divided into four groups (8 in each group) using random digit table: the blank control group treated with distilled water, the test group I treated with Chinese herbal compound of XS (0.135 g/kg), the test group II treated with Chinese herbal compound of HL (0.135 g/kg), and the test group Ill treated with Chinese herbal compound of XS (0.135 g/kg) and HL (0.135 g/kg). All medication was given by gastrogavage once a day for a week. Rats' blood serum was harvested 1 h after the last administration to prepare drug-containing serum. HUVECs were exposed to TNF-alpha (100 ng/mL) to induce cell injury model and incubated with corresponding drug-containing serum (10%) for 24 h. Normal rats' serum was given to cells in the blank control group and the model group, while XC + HL containing serum was given to cells in the rest 3 groups. The adherence of HUVECs and PMN cells was detected by using rose bengal strain. Levels of E-selectin, intercellular adhesion molecule-1 (ICAM-1), and interleukin-1beta (IL-1P) in the supernatant of cultured HU-VECs were determined by ELISA. Protein expressions of mitogen-activated protein kinases p38 (p38MAPK) and extracellular signal-regulated kinase 1/2 (ERK 12) were determined by Western blot.

RESULTSCompared with the blank control group, HUVECs were seriously injured; PMN adherence amount significantly increased; levels of E-selectin, ICAM-1, and IL-1beta increased; expression levels of p-p38MAPK and p-ERK 1/2 in the supernatant of HUVECs significantly increased in the model group (all P < 0.01). Compared with the model group, HUVECs-PMN adherence amount decreased (P < 0.05); levels of E-selectin, ICAM-1, and IL-1 beta in the supernatant of HUVECs decreased (P < 0.01, P < 0.05); expression levels of p-p38MAPK and p-ERK 1/2 of endothelial cells decreased in the test group I, II, and III (P < 0.01).

CONCLUSIONSDrug-containing serums of activating blood, activating blood and dispelling toxin could attenuate TNF-alpha induced injury of HUVECs, inhibit HUVECs-PMN adherence and the release of adhesion factors. Its mechanism might be involved with protein phosphorylation of p38MAPK and ERK 1/2 in the MAPK pathway.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; E-Selectin ; Endothelial Cells ; physiology ; Human Umbilical Vein Endothelial Cells ; Humans ; Inflammation ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-1beta ; Mitogen-Activated Protein Kinase 3 ; Neutrophils ; Rats ; Serum ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

Objective To investigate the effects of CO_2 pneumoperitoneum on the expression of focal adhesion kinase (FAK) of gastric cancer MKN-45 cells. Methods CO_2 pneumoperitoneum with different pressures was simulated in vitro, and the gastric cancer MKN-45 cells were divided into test and control groups. In the test group, gastric cancer MKN-45 cells were cultured in CO_2 pneumoperitoneum with different pressures [5, 10 or 15 mm Hg (1 mm Hg =0.133 kPa)] for 4 hours. The condition of the cells exposed to CO_2 pneumoperitoneum with a pressure of 15 mm Hg was observed at 0.5, 2 and 4 hours. Gastric cancer MKN-45 cells in control group were cultured at normal atmospheric pressure. The expression of FAK and phosphorylated FAK (FAK Tyr397) of each group was detected by Western blot. Multiple-group analysis was done by one-way ANOVA, and intergroup comparison was done by LSD test. Results In CO_2 pneumoperitoneum with pressures of 5, 10, 15 mm Hg, the expression of FAK was 2.14±0.17, 2.07±0.21 and 2.52±0.26, respectively, and the expression of FAK Tyr397 was 1.82±0.28, 1.93±0.52 and 3.71±0.37, respectively. The expression of FAK and FAK Tyr397 in the control group was 2.43±0.46 and 1.71±0.23, respectively. We found significant differences between the 2 groups (F = 2.171, 26.951, P < 0.01). After gastric cancer MKN-45 cells being treated for 0.5, 2 and 4 hours in CO_2 pneumoperitoneum with a pressure of 15 mm Hg, the expression of FAK Tyr397 was 3.41±0.44, 4.12±0.56 and 5.24±0.41 respectively, which is also significantly different (F =116.119, P < 0.01). The expression of FAK Tyr397 was back to 0.72±0.16 1 hour after the release of CO_2. Conclusions CO_2 pneumoperitoneum with different pressures can not promote the expression of FAK in gastric cancer MKN-45 cells which had been cultured for 4 hours, but can activate FAK through promoting its phosphorylation. The degree of FAK phosphorylation increases with pressure and time, and the activity of FAK decreases to pretreatment level rapidly once pressure is released.

OBJECTIVETo explore the viral and host causes of hepatosteatosis in Chinese patient with chronic hepatitis B.

METHODSA total of 562 patients (450 males and 112 females, age range 13-80 years) with biopsy-proven chronic hepatitis B were enrolled in the study. The patients were divided into two groups: group without steatosis (460 patients) and group with steatosis (102 patients). The groups were compared in terms of age, gender, body mass index (BMI), liver enzymes, cholesterol, triglyceride, APO-A, APO-B, urine acid (UA), fasting serum glucose (FSG) and HBeAg, viral load.

RESULTSSteatosis was present in 102 patients (18.15%). The degree of liver steatosis in 97 (95.10%) patients were less 30%. Steatosis was found in 98 (21.78%) of male patients and 4 (3.75%) of female patients (P < 0.01). In the group of chronic hepatitis B with steatosis, the prevalence of obesity, diabetes mellitus, dyslipidaemia, alcoholic consumption, the BMI, cholesterol, triglyceride, UA and FSG levels were significantly higher than those in the group without steatosis (P < 0.01). No significant difference was found in the mean age, HBeAg, viral load between the two groups (P > 0.05). Logistic regression analysis demonstrated that the present of steatosis was positively correlated to BMI, TG and UA.

CONCLUSIONHepatosteatosis in chronic hepatitis B appears to be a result of metabolic factors of the host rather than the effect of viruses.

Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Fatty Liver ; complications ; genetics ; pathology ; virology ; Female ; Hepatitis B virus ; Hepatitis B, Chronic ; complications ; epidemiology ; virology ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Prevalence

OBJECTIVETo study the chemical constituents of Prunella vulgaris.

METHODTo separate the constituents of P. vulgaris by using various kinds of chromatography and identify their structures on the basis of spectral analysis.

RESULTSeven compounds were isolated from the spikes of P. vulgaris. Their structures were established as autantiamide acetate (1), rhein (2), tanshinone I (3), danshensu (4), stigmast-7, 22-dien-3-one (5), 3, 4, alpha-trihydroxy-methyl phenylpropionate (6), butyl rosmarinate (7).

CONCLUSIONCompounds 1-4 were isolated from this genus for the first time.

Amides ; chemistry ; isolation & purification ; Anthraquinones ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; Diterpenes, Abietane ; Flowers ; chemistry ; Lactates ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Phenanthrenes ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Prunella ; chemistry

OBJECTIVETo create Atractylode macrocephala inspissation decoction pieces. The effect of ultrasonic wave on extraction of the active components in A. macrocephala was studied in a water solution.

METHODThe factors including the ratio of material to liquid, ultrasonic power, ultrasonic time, soaking time, particle size etc, were studied. The best extraction method was found through the response surface method.

RESULTThe best extraction method was found as follows: the granularity of material 0.1 mm, the repetition times of ultrasonic process 3 times, the soaking time before the ultrasonic process 30 min, the ratio of liquid to material 10:1, the soaking time after the ultrasonic process 2.6 h, the time of the ultrasonic wave 15.5 min, the power of the ultrasonic wave 531 W, the rate of reservation of active components 88.5%, the rate of inspissation 1.6.

CONCLUSIONThe ultrasonic wave can used in the extraction of the active components in A. macrocephala and a model equation that can be used to predict the experiment was get through the response surface method.

Atractylodes ; chemistry ; Lactones ; isolation & purification ; Oils, Volatile ; chemistry ; isolation & purification ; Particle Size ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; Rhizome ; chemistry ; Sesquiterpenes ; isolation & purification ; Technology, Pharmaceutical ; methods ; Ultrasonics

Acquired Immunodeficiency Syndrome ; Blood Donors ; China ; HIV Infections ; Humans ; Prejudice ; Rural Population

OBJECTIVETo establish the methods of identification and assay in Qianshan Huoxue Gao.

METHODUsing TLC to identify Sanchi, Dragon's Blood and using HPLC to determine the content of ginsenoside Rg1.

RESULTThe linear range of ginsenoside Rg1 was from 0.153 9 to 1.026 microg. The average recovery was 97.4%, RSD was 2.1%.

CONCLUSIONThe methods are simple and have good reproducibility.

Administration, Topical ; Arecaceae ; chemistry ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Ginsenosides ; analysis ; Panax notoginseng ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results

To investigate the differences of proliferation capacity and phenotype properties of mesenchymal stem cells (MSCs) derived from bone marrow (BM) of aplastic anemia patients, fetuses and children, MSCs were isolated from BM of patients with aplastic anemia and expanded in vitro; MSCs derived from BM of fetuses and children were used as normal control groups, three sources of MSCs were compared by morphology, in vitro proliferation capacity, phenotype and immunocytochemistry. The results showed that MSCs could be isolated and expanded from aplastic anemia patient BM. MSCs derived from BM of aplastic anemia patients shared a similar morphology and phenotype with derived MSCs from BM of fetuses and children. However, in vitro proliferation capacity of MSCs derived from BM of aplastic anemia patients after 20 population doublings (PD) was significantly lower, compared with MSCs from BM of fetuses and children. BM MSCs derived from children and fetuses proliferated for more than 30 PD. It is concluded that BM MSCs from aplastic anemia patients appears to be normal in phenotype but their proliferation capacity is lower in comparison with control groups.
Anemia, Aplastic ; blood ; Antigens, CD ; analysis ; Bone Marrow Cells ; cytology ; immunology ; Cell Proliferation ; Child ; Female ; Flow Cytometry ; Humans ; Immunohistochemistry ; Male ; Mesenchymal Stromal Cells ; cytology ; immunology