Burns ; pathology ; Humans ; Wound Healing

A PCR method was used to amplify the sequence encoding the mature peptide of?-mannanase of Bacillus subtilis. The gene was inserted into the Pichia pastoris vector pPIC9K, downstream of?-factor signal peptide sequence. The resultant recombinant plasmid pPIC9K-MAN was lineared by BglII digestion and introduced into the host Pichia pastoris GS115 by PEG method. After screen, the recombinant P. pastoris strain MAN22 was obtained and fermented in large scale 5L fermenter. The recombinant mannanase activity could reach to 1102IU/ml. The properties of the recombinant mannanase were characterized.

To study the effect of cholesterol and 25-OH-cholesterol on cholesterol metabolism in HepG2 cells and the effect of coptisine (Cop) extracted from Coptidis Rhizoma (CR) in reducing and regulating cholesterol. In this study, TC, TG, LDL-c and HDL-c were measured by biochemical analysis; mRNA and protein expressions of LDLR, HMGCR and CYP7A1 were detected by qRT-PCR and Western blot. According to the results, cholesterol and 25-OH-cholesterol inducing could decrease in mRNA and protein expressions of LDLR and CYP7A1, so as to increase TC and LDL-c contents. However, Cop could up-regulate mRNA and protein expressions of LDLR and CYP7A1 and down-regulate that of HMGCR, so as to reduce TC and LDL-c levels. These findings suggested that Cop has potential pharmacological activity for reducing cholesterol, and may reduce cholesterol by regulating mRNA and protein expressions of key genes involved in cholesterol metabolism, such as LDLR, CYP7A1 and HMGCR. This study laid a firm theoretical foundation for developing new natural drugs with the cholesterol-lowering activity.
Berberine ; analogs & derivatives ; pharmacology ; Cholesterol ; metabolism ; Cholesterol 7-alpha-Hydroxylase ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Enzymologic ; drug effects ; Hep G2 Cells ; Humans ; Hydroxymethylglutaryl CoA Reductases ; genetics ; metabolism ; Receptors, LDL ; genetics ; metabolism ; Triglycerides ; metabolism

It designs a way that can easily screen high-pyruvate-producing strain.It is a intelligently selected method which can highly improve the efficiency of strain screening.The principle can be described as the following:On the CaCO-3 medium,a transparent ring can be exhibited based on the reaction of PYR produced by the strain and CaCO-3 in the medium for pyruvate-calcium is a kind of soluble substance,it is obviously that the high-pyruvate-producing strain has a bigger dimension of the transparent ring.On the other hand,color reaction between TTC and ADH indicate the enzyme activities which have a proportional relation with color,our object strain is a weak-ADH-enzyme-activities type with a weak metabolic flux from PYR to alcohol.So the white color strain may be the right choice.

OBJECTIVETo observe the changes in matrix metalloproteinase-2,7 (MMP-2,7) and tissue inhibitor of metalloproteinase-2 (TIMP-2) in deep partial thickness burn during the process of wound healing, and the effects of bFGF on wound healing.

METHODSThe rats inflicted by 30% TBSA deep partial thickness burn were randomly divided into simple scald and bFGF treatment groups. Biopsies from wound skin were harvested at 3 and 6PBHs and 1, 3, 7, 14 PBDs for the detection of the epithelialization rate and collagen content. The above indices were also detected in the skin of another 6 normal rats as normal control.

RESULTS(1) The epithelialization rate in bFGF treatment group was higher than that in simple scald group during 3PBH to 14 PBD. (2) The collagen contents in both bFGF treatment group and simple scald group were continually decreased during 3 PBH to 3 PBD, and increased from 7 to 14 PBD, but still lower than that in normal control (P < 0.05). (3) The expression of MMP-2,7 and TIMP-2 in simple scald group enhanced from 1 to 14 PBD, and peaked on 7 PBD. (4) The expression of MMP-2,7 in bFGF treatment group was similar to that in simple scald group from 3 to 6 PBH, while the expressions of MMP-2,7 and TIMP-2 was higher than those in simple scald group from 1 to 14 PBD.

CONCLUSIONThe collagen deposition would be affected by the activities of extracellular matrix in scald wound in rats. Changes in MMP-2,7 and TIMP-2 expressions were an important process of wound repair, which was closely related to the acceleration of wound healing by the application of bFGF.

Animals ; Burns ; metabolism ; Collagen ; analysis ; Epithelium ; physiology ; Fibroblast Growth Factor 2 ; pharmacology ; Male ; Matrix Metalloproteinase 2 ; analysis ; Matrix Metalloproteinase 7 ; analysis ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-2 ; analysis ; Wound Healing

OBJECTIVETo observe the expression characteristics of EGFR and FGFR-2 in normal skin from fetal and adult, and attempted to probe the molecule mechanism of fetal scarless healing.

METHODSThe skin samples of fetal and adult were taken from abortive fetus of obstetrics unit and donor site of plastic operation patients in our burn unit, respectively. EGFR and FGFR-2 were used as the biochemical markers for reparative cells. Immunohistochemistry staining technique was employed to determine the expressive levels of different epithelial cells markers.

RESULTSThere were EGFR and FGFR-2 antibody positive cells in normal skin from fetal and adult, but the expressive levels of EGFR and FGFR-2 protein had apparent difference, with the time of fetation increasing, the EGFR and FGFR-2 positive expression rate became stronger gradually. The number of FGFR-2 antibody positive cells found in adult skin was much more than that in fetal skin.

CONCLUSIONThere were the inherent differences of EGFR and FGFR-2 antibody immunohistochemistry staining in cells of adult and fetal skin. which may be an essential facet of fetal scarless healing.

Adult ; Epithelial Cells ; metabolism ; ErbB Receptors ; metabolism ; Female ; Fetus ; metabolism ; Humans ; Immunohistochemistry ; Receptor, ErbB-2 ; metabolism ; Skin ; metabolism ; Wound Healing

Objective To evaluate serum soluble intercellular adhesion molecule-1 (sICAM-1) before and after lung transplantation for diagnosing acute rejection. Methods Biotin-streptavidin time resolved fluoroimmunoassay (BSA-TRFIA) was used to detect the concentration of serum sICAM-1 before and after lung transplantation in 26 patients. All patients were divided into stable lung transplantation group (n =16), acute rejection group (n =4) and infected group (n =6). The serum level of sICAM-1 in those groups was compared with that of the control group ( n = 30 ) by the non-parametric rank sum test ( KruskalWallis H test). Results No significant difference was found for serum sICAM-1 among the three groups and the control group before operation: (357.07 ± 220.01 ), ( 396. 18 ± 136.25 ), (468.95 ± 85.48 ) μg/L vs(348.63 ±69. 12) μg/L, H=6. 0436, P ＞0.05. However, when rejection and infection happened after operation, the serum sICAM-1 increased in the acute rejection group (455.53 ± 126.51 μg/L) and decreased in the infection group (146.43 ± 327.11 μg/L), and the level in the stable transplantation group was (274.23 ± 157.53 ) μg/L (H = 21. 8994, P ＜ 0.01 ). Conclusion Serum sICAM-1 level might be a potential marker to differentiate acute rejection from infection after lung transplantation.

OBJECTIVETo investigate the impact of hCG on the epidermal growth factor (EGF) in the phallic tissues of hypospadiac mice.

METHODSThe Kunming mouse model of congenital hypospadias was established. As controls, 10 three-week-old normal male mice and 10 3-week-old congenital hypospadiac mice were injected with isometric sodium chloride intraperitoneally. Another 40 mice with hypospadias were divided into 4 groups and injected intraperitoneally with 50 IU, 100 IU, 150 IU and 200 IU of hCG respectively per day for 7 days. Then the concentration of ECG in the phallus and in the serum were assayed by ELISA.

RESULTSThe EGF level in the phallus of the hypospadiac mice was much lower than that of the controls statistically (P < 0. 05). After the injection of exogenous hCG, the concentration of ECG increased with the dose of hCG, 50 IU (59.57 +/- 22.16) pg/ml, 100 IU (57.97 +/- 9.59) pg/ml, 150 IU (73.02 +/- 31.35) pg/ml and 200 IU (92.45 +/- 35.74) pg/ml. The concentration of ECG showed no significant difference between the control group and the 150 IU and 2000 IU groups (P > 0.05).

CONCLUSIONDecreased concentration of EGF in the mouse phallus may be associated with the pathogeny of hypospadias. A proper dose of exogenous hCG may increase the concentration of EGF in the mouse phallus.

Animals ; Chorionic Gonadotropin ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay ; Epidermal Growth Factor ; biosynthesis ; blood ; Hypospadias ; blood ; metabolism ; Injections, Intraperitoneal ; Male ; Mice ; Penis ; drug effects ; metabolism ; pathology

To investigate the practicability of establishing zebrafish lipid-lowering drug screening model and the effect of berberine (BBR) on hyperlipidemic zebrafish. Three-month-old zebrafishes were fed with 4% cholesterol for 0, 2, 4, 8, 14, 20, 25, 30 days, and the level of total cholesterol in serum was measured. Zebrafish were randomly divided into four groups: the control group, the high cholesterol diet group, the 0.01% simvastatin-treated group, the 0.1% berberine-treated group and the 0.2% berberine-treated group. The levels of total cholesterol (TC), triglyceride (TC), low density lipoprotein cholesterol (LDL-c) and high-density lipoprotein cholesterol (HDL-c) in serum were measured; the expression of hepatic HMGCR, LDLR and CYP7A1a mRNA expressions were detected by real time PCR. Oil red O staining was performed to observe the changes in fat content in the liver. According to the result, the level of serum TC in the 4% cholesterol diet group significantly was higher than that of the normal control group in a time-dependent manner and reached a stable level at the 20th day. The BBR group showed significant decreases in the levels of TC, TG and LDL-c, HMGCR mRNA expression and fat content and increases in LDLR and CYP7A1a mRNA. The hyperlipidemia zebrafish model was successfully established by feeding with 4% cholesterol for 20 days. The findings lay a foundation for further screenings on lipid-lowering drugs.
Animals ; Berberine ; administration & dosage ; Cholesterol ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Hyperlipidemias ; drug therapy ; metabolism ; Hypolipidemic Agents ; administration & dosage ; Liver ; drug effects ; metabolism ; Male ; Triglycerides ; metabolism ; Zebrafish ; metabolism

OBJECTIVETo study the feasibility and cost-effectiveness of a case-finding program on tuberculosis (TB) in richer rural areas.

METHODSScreening was implemented every three months for a total period of 9 months, in rural areas with high case notification rates. Three villages, each with ten thousand population, were selected to carry out a household screening program. A suspect was defined as who coughed for more than 3 weeks. The suspect was then referred to further diagnosis in county TB dispensary to undergo chest X-ray and sputum test.

RESULTSOf the 86,168 community population screened, 26 TB patients were identified with 7 of them were smear positive. The ratio of effectiveness vs. cost decreased on the second but slightly increased on the third screening program. The direct costs for the 3 screening programs were 6,312,397 and 1637 RMB respectively. Of total direct cost, 5.9% was paid by TB patients, whereas 35.9% was through financing of the county itself.

CONCLUSIONThe community household screening program could achieve higher case detection rate than passive case-finding approach which could be used in richer areas with low case detection rate in China.

China ; Chronic Disease ; Cost-Benefit Analysis ; Cough ; etiology ; Family Characteristics ; Humans ; Mass Screening ; economics ; Radiography, Thoracic ; Rural Health ; Sputum ; microbiology ; Tuberculosis ; complications ; diagnosis