Effect of Jingfang Granules on carrageenan-induced tail thrombosis in mice based on ERK/p38 MAPK signaling pathway / 中国中药杂志

The present study explored the anti-inflammatory and anti-thrombotic mechanism of Jingfang Granules on tail thrombosis induced by carrageenan in mice. Thirty-two male ICR mice were randomly divided into a control group, a model group, a Jingfang Granules group, and a positive drug(aspirin) group, with eight mice in each group. The thrombosis model was induced by intraperitoneal injection of carrageenan(45 mg·kg~(-1)) combined with low-temperature stimulation, and the mice were treated with drugs for 7 days before modeling. Twenty-four hours after modeling, blood was detected for four blood coagulation indices in each group. The enzyme-linked immunosorbent assay(ELISA) was used to detect the activity of plasma interleukin-6(IL-6), interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and other inflammatory factors. The tails of mice in each group were cut off to observe tail lesions and measure the length of the thrombus. The protein expression and phosphorylation level of extracellular signal-regulated kinase 1/2(ERK1/2) and p38 mitogen-activated protein kinase(p38 MAPK) in spleen tissues were detected by Western blot. The results showed that dark red thrombus appeared in the tails of mice in each group. The length of the black part accounted for about 40% of the total tail in the model group. Additionally, the model group showed prolonged prothrombin time(PT), increased fibrinogen(FIB) content, and shortened activated partial thromboplastin time(APTT). Compared with the model group, the groups with drug intervention displayed shortened black parts in the tail and improved four blood coagulation indices(P<0.05). As revealed by ELISA, the expression levels of TNF-α, IL-1β, and IL-6 in the mouse plasma were significantly up-regulated in the model group, and those in the groups with drug intervention were reduced as compared with the model group(P<0.05). As demonstrated by Western blot, the protein expression and phosphorylation levels of ERK1/2 and p38 MAPK in the spleen tissues were significantly elevated in the model group, while those in the Jingfang Granules group were down-regulated as compared with the model group with a significant difference. Jingfang Granules can inhibit tail thrombosis of mice caused by carrageenan presumedly by inhibiting the activation of ERK1/2 and p38 MAPK signaling pathways.

3-Dimensional model reconstruction of penis and surrounding tissue / 中华整形外科杂志

<p><b>OBJECTIVE</b>To evaluate the feasibility of 3-Dimensional (3-D) model reconstruction of penis and surrounding structures based on magnetic resonance images, which may provide the model building method for modeling surgery of individual penoplasty.</p><p><b>METHODS</b>Magnetic resonance (MR) images of penis with different imaging parameters were evaluated. With the surface rendering construction, the 3D virtual model was established by Amira software.</p><p><b>RESULTS</b>The anatomical details imaging is better in T2-weighted fast spin-echo images with 3.0 mm slice thickness. The established model based on the MR images can show the soft-tissue, suspensory ligament of the penis. The suspensory ligament stretches between the pubic symphysis and the corpora cavernosa. The penile roots attach to inferior ramus of pubis.</p><p><b>CONCLUSIONS</b>MR imaging provides enough anatomical information for modeling. It can be used for the development of model surgery system of individual penoplasty.</p>

The three-dimensional reconstruction of penile suspensory ligament and adjacent structures based on the MRI image / 中华整形外科杂志

<p><b>OBJECTIVE</b>To establish a three-dimensional image of the penile suspensory ligament, and explore a stereoscopic and multi angle observation method of patient' s penile suspensory ligament.</p><p><b>METHODS</b>This study selected the patients with small penis from our hospital as subjects. The participants were conducted on magnetic resonance imaging (MRI) examination before operation. Afterwards, the results of MRI were imported into 3D reconstruction software (MIMICS 10.0), and the suspensory ligament of penis, the pubic symphysis and other related structures were reconstructed for observation.</p><p><b>RESULTS</b>The pubic symphysis, penis and corpus spongiosum can be quite clearly displayed in the thin-section MRI images. In addition, penile suspensory with patchy distribution can be visible between lower part of ligament pubic symphysis and corpus cavernosum. Finally, we can reconstruct the three-dimensional structures through MIMICS 10.0, and then precisely describe the suspensory ligament's start-stop point, the angle with cavernous body of penis and the attached area in the corpus cavernosum penis.</p><p><b>CONCLUSION</b>Based on the MRI 3D reconstruction of deep penile suspensory ligament and adjacent structures, we can carry out dynamic, three-dimensional multi angle observation of patients deep penile suspensory ligament, and can use the reconstructed image to provide certain theory basis for the judgement of the corpus cavernosum penis extension length and penile suspensory ligament depth before penis extension operation.</p>

Modulatory effect of Rac1 protein on epidermal stem cells migration during wound healing / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate modulatory role of Rac1 protein in epidermal stem cell (ESC) migration during wound healing, in order to provide a reference for enriching basic theory of wound healing and guiding clinical application.</p><p><b>METHODS</b>Constitutively active mutant of Rac1 protein (Rac1Q61L) or dominant negative isoform of Rac1 protein (Rac1T17N) was transfected into ESC using a retroviral vector FUGW, and retroviral vector FUGW transfected into ESC in singles was used as blank control. The cells were divided into 3 parts according to the random number table and treated as follows. First, equal numbers of cells were inoculated into 24-well plates coated with collagen I (20 µg/mL), collagen IV (20 µg/mL) or fibronectin (10 µg/mL). Cells adhered to above matrices were quantitated using CytoTox 96 colorimetric kit. Second, 1000 cells adhered to collagen IV, after being stained with tetramethyl rhodamine isothiocyanate-phalloidin, were collected for observation of cell morphology and comparison of spreading area under confocal laser scanning microscope. Third, ESC with density of 2 × 10(5) cells per well were placed in upper compartment of Transwell chamber, DK-SFM culture medium alone or that containing stromal cell derived factor 1 (SDF-1) was added into lower compartment of Transwell chamber. Migration of ESC was observed using inverted phase contrast microscope, and the result was denoted as migration rate. Lastly, ESC with density of 7.5 × 10(5) cells per well was inoculated into 6-well plates for 12 hours, and treated with 4 µg/mL mitomycin C for 2 hours. The remaining scratch width of monolayer was respectively measured 6 hours or 12 hours after scratching to calculate the percentage of remaining scratch width. Data were processed with t test.</p><p><b>RESULTS</b>Compared with that of blank control, the number of Rac1Q61L-transfected cells adhered to collagen I was significantly increased (t = 5.302,P < 0.05), while the number of Rac1T17N-transfected cells adhered to collagen I, IV, and fibronectin were all obviously decreased (with t value respectively 13.741, 15.676, 8.256, P values all below 0.05). Confocal laser scanning microscope showed that spreading area of Rac1Q61L-transfected ESC (with laminate pseudopodia on edge) and Rac1T17N-transfected ESC was respectively larger and smaller as compared with that of blank control. With SDF-1 effect, the migration rate of Rac1T17N-transfected ESC was decreased by 78.0% and Rac1Q61L-transfected ESC was increased by 43.4% as compared with that of blank control. Without SDF-1 effect, the migration rate of Rac1T17N-transfected ESC was decreased by 55.2%, while the migration rate of Rac1Q61L-transfected ESC was close to that of blank control. Six or 12 hours after scratching, the percentage of remaining scratch width in Rac1Q61L-transfected ESC was lower as compared with that in blank control [(39 ± 9)% vs. (43 ± 5)%, (6 ± 5)% vs. (18 ± 7)%, with t value respectively 1.027, 4.389, with P value respectively above and below 0.05], while that in Rac1T17N-transfected ESC [(81 ± 9)%, (71 ± 11)%, respectively] was obviously higher as compared with that in blank control (with t value respectively 11.386, 11.726, P values all below 0.05).</p><p><b>CONCLUSIONS</b>Rac1 protein may control the migration of ESC by regulating its adhesion, spreading, and chemotaxis, and it plays an active role in wound healing accelerated by ESC.</p>

A model of individually forecasting the penile length gained after penis lengthening / 中华整形外科杂志

<p><b>OBJECTIVE</b>To establish a model of individually forecasting the penile length gained after penis lengthening.</p><p><b>METHODS</b>A total of 322 patients were diagnosed congenital penile shortness and received partial suspensory ligament release in our department from Oct. 1988 to Apr. 2011. The patients were divide into two groups as Modeling Group (n = 200) and Checking Group (n = 122). Then a two-dimensional model of the suspensory-ligament-release penis lengthening is established. In the Modeling Group, a statistical analysis of the penile length in flaccid and erectile state before and after penis lengthening was carried out, and a forecasting predictive function of increased penile length was derived. Then the predictive accurate rate was tested in the Checking Group.</p><p><b>RESULTS</b>There was a significant linear correlation between the increased length in flaccid and erectile state (correlation coefficient = 0.921, P < 0.01), there was also a similar relationship between the extension rate of erection before and after the operation (correlation coefficient = 0.803, P < 0.01). According to the significant linear correlations showing above, two regression forecasting models were established. A predictive function of increased flaccid length was derived from the two regression forecasting models. The effectively forecasting rates were 84.5% (169/200, the Modeling Group) and 87.7% (107/122, the Checking Group) when the absolute value of forecasting error was less than 1.5 cm.</p><p><b>CONCLUSIONS</b>The discovered significant correlation and the established forecasting function provide us a model of roughly and individually forecasting the penile length gained after penis lengthening.</p>

The cellular and molecular mechanism of inhibitory effect of asiaticoside on capsular contracture following breast augmentation / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the cellular and molecular mechanism of the inhibitory effect of asiaticoside on capsular contracture following breast augmentation.</p><p><b>METHODS</b>Contracture capsule derived fibroblasts were cultured in medium with different concentration of asiaticoside. The cell proliferation, collage synthesis and alpha-SMA expression were detected by means of 3H-thymidine incorporation, 3H-proline incorporation, and Western-blot. The results were analyzed by SPSS 11.0 with t test.</p><p><b>RESULTS</b>DNA and collagen synthesis of fibroblasts were dramatically inhibited when the asiaticoside reached the concentration of 50 mg/L. The inhibitory rate was 34.7% and 30.1% respectively, showing a significant difference from that in control group (P < 0.05). The inhibitory effect increased with the rise of the asiaticoside concentration in a dose-dependent manner. When the concentration of asiaticoside reached 25 mg/L, the expression of alpha-SMA was down-regulated with an activation index of 1.673, showing a significant difference when compared with that in control group (P < 0.05).</p><p><b>CONCLUSIONS</b>asiaticoside can effectively inhibit the DNA and collagen synthesis of capsule-derived fibroblasts. The trans-differentiation of fibroblast to myo-fibroblasts is also prevented by it.</p>

Expression of cyclooxygenase-2 and its relationship with mismatch repair and microsatellite instability in hereditary nonpolyposis colorectal cancer.

OBJECTIVE: To investigate cyclooxygenase-2 (COX-2) expression and its relationship with mismatch repair (MMR) protein expression and microsatellite instability (MSI) in hereditary nonpolyposis colorectal cancer (HNPCC). METHODS: A total of 28 cases of colorectal adenoma and 14 cases of colorectal carcinoma were collected between July 2003 and July 2007 from 33 HNPCC families. Sporadic colorectal adenoma (n=32) and carcinoma patients (n=24) served as controls. With samples of tumor tissues and normal colonic mucosa collected from the patients, the protein expressions of COX-2 and MMR (hMLH1, hMSH2, and hMSH6) were examined with immunohistochemical assay. Frequency of MSI in five standard MSI loci BAT25, BAT26, D2S123, D5S346, and D17S250 were analyzed by means of polymerase chain reaction. RESULTS: The rate of COX-2 high-expression was 53.6% (15/28) and 42.9% (6/14) in HNPCC adenoma and carcinoma; 62.5% (20/32) and 91.7% (22/24) in sporadic adenoma and carcinoma, respectively. That rate was lower in HNPCC carcinoma than in sporadic carcinoma (Pü0.05). MMR-deletion rate and percentage of high-frequency MSI (MSI-H) in HNPCC carcinoma were higher than those in sporadic colorectal carcinoma [both 71.4% (10/14) vs. 12.5% (3/24), both Pü0.01]. Among the 10 MMR-deficient HNPCC carcinoma patients, COX-2 low-expression was observed in 8 cases (80.0%), while COX-2 high-expression was observed in all of the 4 MMR-positive HNPCC carcinoma cases (Pü0.05). In comparison to MMR positive HNPCC carcinoma, HNPCC adenoma, and sporadic carcinoma, COX-2 expression was significantly lower in corresponding MMR-deficient cases (all Pü0.05). The rates of COX-2 low-expression in HNPCC adenoma, HNPCC carcinoma, and sporadic carcinoma with MSI-H were significantly higher than those in the cases with microsatellite stability (all Pü0.05). CONCLUSION: COX-2 is expressed at a low level in HNPCC carcinoma, different from the high COX-2 expression in sporadic carcinoma.

Clinicopathological features of non-familial colorectal cancer with high-frequency microsatellite instability.

OBJECTIVE: To explore the clinicopathological features of non-familial colorectal cancer with high-frequency microsatellite instability (MSI-H). METHODS: One hundred and fifty patients with colorectal cancer who had no family history were enrolled in this study from June 2006 to June 2008. Five standard microsatellite loci including BAT25, BAT26, D2S123, D5S346, and D17S250 were amplified with immunofluorescent polymerase chain reaction. The patient information including age, sex, and tumor location was recorded. Pathological features including differentiation, mucinous differentiation, histological heterogeneity, and Crohn's-like reaction were observed under light microscope. The presence of tumor-infiltrating lymphocytes (TLs, CD4+ and CD8+) was detected by means of immunohistochemistry. A regression equation was obtained by stepwise logistic regression analysis to evaluate the relationship between MSI-H phenotype in colorectal cancer and pathological features. RESULTS: MSI-H phenotype occurred in 13.33% of the 150 patients with non-familial colorectal cancer. Poor differentiation, histological heterogeneity, Crohn's-like reaction, and presence of TLs were found to be independent factors to identify MSI-H non-familial colorectal cancer. Logistic regression equation showed an overall sensitivity of 70.0%, specificity of 99.2%, and accuracy of 95.3% in identifying MSI-H non-familial colorectal cancer. CONCLUSION: MSI-H non-familial colorectal cancer manifests specific pathological features, which may be relied upon for effective identification of that disease.

Promotion effect of stromal cell-derived factor 1 on the migration of epidermal stem cells in the healing process of frostbite-wound model ex vivo / 中华烧伤杂志

<p><b>OBJECTIVE</b>To study the promotion effect of stromal cell-derived factor 1 (SDF-1) on the migration of epidermal stem cells (ESC) in the healing process of frostbite-wound model ex vivo.</p><p><b>METHODS</b>A three-dimensional model of full-thickness frostbite of skin was constructed (with slot-like wound) out of skin equivalent. The expression of SDF-1 in wound stroma was observed with immunohistochemistry staining on post injury days (PID) 3 and 7. The model frostbite wounds were divided into control group (treated with PBS 50 microL per wound), SDF-1 group (treated with 100 ng/mL SDF-1, 50 microL per wound), and AMD3100 group [treated with 100 ng/mL AMD3100 (50 microL per wound) for 30 minutes, and then SDF-1 50 microL was added per wound]. The redistribution of ESC around wound was observed.</p><p><b>RESULTS</b>The expression of SDF-1 in wound stroma increased gradually on PID 3 and 7. Compared with those in control and AMD3100 groups, there were more ESC and epithelial cell layers, and more integrin beta(1)-positive cells appeared at the basal layer of wound in SDF-1 group, and some of the positive cells migrated upward to epidermis.</p><p><b>CONCLUSIONS</b>SDF-1 contributes to wound repair through promoting ESC to migrate toward and gather around wound edge. This may be one of the mechanisms of ESC participating in wound repair.</p>

Effect of staphylococcal enterotoxin C injection on post liposuction seroma / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the mechanism of treatment of post liposuction seroma with Staphylococcal enterotoxin C injection.</p><p><b>METHODS</b>64 cases with post liposuction seroma were treated with Staphylococcal enterotoxin C injection or routine procedures. The exudate of those patients was collected to analyze the ratio, pH value, cell species and numbers, and the value of TP, ALP, LDH, AST, ALT, gamma-GT, ADA, ApoB, TC.</p><p><b>RESULTS</b>The ratio, numbers of lymphocyte and mesothelial cells and TP, LDH, ADA, TC value in exudate in Staphylococcal enterotoxin C group were significantly higher than those in control group.</p><p><b>CONCLUSIONS</b>The effect of Staphylococcal enterotoxin C injection on the exudate of seroma may be related to the non-inflammation reaction.</p>

Significance of the secreted frizzled-related protein 2 expression in earlobe keloid / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the expression and the role of secreted frizzled-related protein 2 (SFRP2) in the earlobe keloid and find a valid way to treat the keloid with gene therapy.</p><p><b>METHODS</b>The expression of SFRP2 mRNA and protein was tested with in situ hybridization and Western Blot Analysis method in the different period of earlobe keloid.</p><p><b>RESULTS</b>The SFRP2 mRNA and protein expression at the keloid edge was significantly high in 12 month group than in 3 or 6 month groups (P < 0.01), but not than in 24 month group. The SFRP2 expression started to decrease in the keloid center 12 month later (P < 0.01). The SFRP2 expression was always higher in edge than in center during all the period (P < 0.05, P < 0.01).</p><p><b>CONCLUSIONS</b>The results suggest that SFRP2 may play an important role in the development of keloid, especially at the keloid edge. The high SFRP2 expression in endothelial cells and surrounding tissue is also important. It may be a new way for gene therapy of keloid by decreasing the SFRP2 expression.</p>

[Application of ICP-AES to detecting nutrients in grain of wheat core collection of China].

Deficiency of micronutrients, especially iron and zinc, has been a serious malnutrition problem worldwide in human health. Increasing Fe and Zn concentrations in grains by means of plant breeding is a sustainable, effective and important way to improve human mineral nutrition and health. However, little information on grain Fe and Zn concentrations in Chinese wheat genotypes is available. Therefore, to determine the nutrients status especially these of micronutrients in wheat grain is necessary and very useful. Two hundred sixty two genotypes were selected from the wheat mini-core collections, which contained 23090 wheat genotypes in China and represented 72.2% of total genetic variation. All 262 genotypes were grown in soils of similar geographical and climate location in order to minimize the environmental effect. After harvesting, the grains were washed with deionized water and dried (around 70 degrees C), then digested in HNO3 solution using a microwave accelerating reaction system (MARS). Nutrient concentrations in stock solution were analyzed by inductively coupled plasma atomic emission spectrometry (ICP-AES). Remarkable genetic variations among grain nutrient concentrations (Fe, Mn, Cu, Zn, Mg, Ca, K and P ) in the tested genotypes were detected. The concentrations of Fe, Zn, Mn, Cu, Ca, Mg, K and P in wheat grain were in the ranges of 34.2-61.2, 26.3-76.0, 20.9-56.7, 3.4-9.8, 290-976, 1129-2210 mg x kg(-1); 0.34%-0.85% and 0.296%-0.580%, respectively. The corresponding average values were 45.1, 50.2, 37.9, 6.5, 515, 1772 mg x kg(-1), 0.55% and 0.451%, respectively. Significant positive correlations between micronutrients (Fe, Mn, Zn, and Cu) in wheat grains were detected, and the correlation coefficients were 0.395** (Fe and Mn), 0.424** (Fe and Zn), 0.574** (Fe and Cu), and 0.474** (Mn and Cu), respectively. However, no significant difference was found in grain nutrient concentrations between spring-wheat and winter-wheat genotypes. This study provides valuable and important information for breeding wheat genotypes which are enriched with minerals in grains, especially Fe and Zn

To transdifferentiate human hypertrophic scar fibroblasts induced by connective tissue growth factor mediated transforming growth factor-beta 1 in vitro / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the role of connective tissue growth factor (CTGF) induced TGF-beta 1 in the transdifferentiation of human hypertrophic scar fibroblast (HSFb).</p><p><b>METHODS</b>Human hypertrophic scar fibroblasts were cultured in vitro, 5 cell samples were stimulated with TGF-beta 1 (0, 2.5, 5.0, 7.5, 10.0 ng/mL, respectively) for 48 hours; other cell samples were divided into: normal control (NC) group, CTGF group (with addition of 10 ng/mL rhCTGF into culture medium), TGF-beta 1 group (with addition of 10 ng/mL TGF-beta 1 into culture medium), CTGF ASODN group (with addition of 10% FBS-DMEM after transfection of CTGF ASODN), CTGF ASODN + TGF-beta 1group (with addition of 10 ng/mL TGF-beta 1 after transfection of CTGF ASODN). Expression of CTGF was determined by Western blotting with stimulation of different concentration of TGF-beta 1. Expression of alpha-smooth muscle actin (alpha-SMA) was measured by Western blotting. Positive cell rate of alpha-SMA was examined by flow cytometry.</p><p><b>RESULTS</b>With stimulation of 10.0 ng/mL TGF-beta 1, the expression of CTGF was obviously higher than that of non-stimulation (P < 0.05). Expression of alpha-SMA in the CTGF group and the TGF-beta 1 group was obviously higher than that in NC group (P < 0.01), while there was no obvious difference among NC, CTGF ASODN, CTGF ASODN + TGF-beta 1 groups (P > 0.05). The positive cell rate of alpha-SMA in NC, CTGF, TGF-beta 1, CTGF ASODN, CTGF ASODN + TGF-beta 1 groups was (10.8 +/- 2.8)%, (29.1 +/- 4.0)%, (28.7 +/- 4.8)%, (10.7 +/- 2.3)%, (14.3 +/- 2.9)%, respectively, which was similar to expression of alpha-SMA on statistic analysis.</p><p><b>CONCLUSIONS</b>CTGF is one of the most important downstream efforts for TGF-beta 1 in inducing the transdifferentiation of HSFb.</p>

The interaction between secreted frizzled-related protein 2 and osteoblast-specific factor 2 in keloid / 中华烧伤杂志

<p><b>OBJECTIVE</b>To verify the interaction between secreted frizzled-related protein 2 (SFRP2) and osteoblast-specific factor 2 (OSF-2).</p><p><b>METHODS</b>HA-tagged OSF-2 fusion protein recombinant vector pCMV-HA-OSF-2, which could express in mammal cells was constructed, then identified by enzyme-cutting and transfected into human kidney 293 (HK293) cells with or without Myc-SFRP2 recombinant eukaryotic expression vector pCMV-HA-SFRP2. The interaction between SFRP2 and OSF-2 was detected through coimmunoprecipitation and Western blotting.</p><p><b>RESULTS</b>In electrophoresis bath, target fragment of SFRP2 coding gene with 800 bp and target gene OSF-2 with 2500 bp could be seen respectively after enzyme-cutting, which showed that pCMV-Myc-SFRP2 and pCMV-HA-OSF-2 were constructed successfully. No HA-OSF-2 expression was detected after pCMV-Myc-SFRP2 or pCMV-HA-OSF-2 transfection. Whereas, HA-OSF-2 expressed by Myc antibody immunoprecipitation after pCMV-Myc-SFRP2 and pCMV-HA-OSF-2 co-transfection.</p><p><b>CONCLUSIONS</b>HA-OSF-2 recombinant vector can express in mammal cells. Interaction exists between HA-OSF-2 and SFRP2.</p>

Effects of genistein on expression of TGF-beta1 and intracellular free Ca2+ concentration in fibroblasts derived from human hypertrophic scar / 中华整形外科杂志

<p><b>OBJECTIVE</b>To observe the effects of Genistein on TGF-beta1 expression and the intracellular free Ca2+ concentration in human hypertrophic scar fibroblasts, and to discuss the mechanism of the anti-fibrosis effect.</p><p><b>METHODS</b>Fibroblasts were derived from human hypertrophic scar tissue and cultured in vitro. Genistein in different concentrations (25, 50, 100 micromol/L) was administrated to the fibroblasts, respectively. After 48 hours of co-culture, the expression of TGF-beta1 mRNA and protein were examined by RT-PCR and Western-Blot assay respectively. The intracellular free Ca2+ concentration in hypertrophic scar fibroblasts pretreated by Genistein was determined by laser confocal scanning microscopy with or without the stimulation of bFGF.</p><p><b>RESULTS</b>Genistein inhibited the expression of TGF-beta1 in hypertrophic scar fibroblasts on a concentration-dependent manner. bFGF significantly elevated the intracellular free Ca2+ concentration, however its stimulating effect was remarkably alleviated when the fibroblasts were pre-treated by Genistein.</p><p><b>CONCLUSIONS</b>Genistein can reduce the expression of TGF-beta1 and block the accumulation of intracellular free calcium induced by growth factors. It maybe one of the possible mechanisms of Genistein's antifibrosis effect.</p>

The effects of genistein on tyrosine protein kinase-mitogen activated protein kinase signal transduction pathway in hypertrophic scar fibroblasts / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the inhibitory effects of genistein on tyrosine protein kinase (TPK)-mitogen activated protein kinase (MAPK) signal transduction pathway in hypertrophic scar fibroblasts (HSFb), in order to explore the molecular mechanism of inhibition of scar hyperplasia by genistein.</p><p><b>METHODS</b>HSFbs were isolated from human hypertrophic scar tissues and cultured in vitro. The cells were treated by genistein in different concentrations (25, 50, 100 micromol/L, respectively), followed by basic fibroblast growth factor (bFGF) stimulation. The activity of TPK was assessed with [gamma-32P] ATP substrate incorporation. The phosphorylation protein expression levels of main signal molecules in TPK-Ras-MAPK pathway including c-Raf, MEKl/2, extracellular signal regulated kinase (ERK), p38MAPK, c-Jun N-terminal kinase (JNK) were determined by Western blot. HSFbs treated with dimethylsulfoxide (DMSO) were used as control group.</p><p><b>RESULTS</b>After being treated with genistein in concentration of 25, 50, 100 micromol/L, the activity of TPK in HSFbs was depressed significantly [(7.15 +/- 0.35) x 10(5), (5.62 +/- 0.88) x 10(5), (5.62 +/-0.88) x 10(5) 10(5) pmol x min(-1) x mg(-1), respectively] when compared with that in control group [(8.92 +/- 0.28) x 10(5) pmol x min(-1) x mg(-1), P < 0.05]. Compared with those in control group,the phosphorylation protein expression levels of c-Raf, MEK1/2, ERK1/2 and p38 MAPK were lowered in different degree (P < 0.05 or P < 0.01) after genistein treatment. The phospho-JNK levels after treatment with genistein were similar to that of control group. Under the condition of pretreatment with genistein, the activities of TPK and signal pathway protein expressions in HSFb showed a downward trend after stimulation with bFGF.</p><p><b>CONCLUSION</b>Genistein can effect the proliferation and activation of HSFb by inhibiting the phosphorylation of receptor of TPK signal transduction pathway (TPK --> Raf --> MEK --> ERK/p38).</p>

Effect of qishen huoxue granule combined with fluconazole on survival rate of mice with systemic C. albaicans infection / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Qishen Huoxue Granule (QHG) combined with Fluconazole on the survival rate of mice with systemic C. albaicans (CA) infection.</p><p><b>METHODS</b>Deep CA infection model mice, with normal and low immunity, were established separately by injecting standard strain of CA via caudal vein, and were divided into 4 groups at random, treated by gastrogavage with normal saline (Group A), QHG (Group B) Fluconazole (FCZ, Group C) and QHG + FCZ (Group D) respectively, and a blank group was set up with normal mice for control. The survival time and the total survival rate in 30 days in various groups were recorded.</p><p><b>RESULTS</b>For mice with normal immunity, the survival rate in Group D and C was 79% and 78% respectively, showing no difference between them (P > 0.05). But for those with low immunity, it was 36% and 7% respectively, and the survival rate significantly higher in Group D than in Group C (P < 0.05).</p><p><b>CONCLUSION</b>As compared with those treated with FCZ alone, QHG combined with FCZ can raise the survival rate of the immuno-suppressed mice with systemic CA infection.</p>

Cloning and Expression of the Full-length cDNAs Encoding Human Class Ⅰ Alcohol Dehydrogenases / 现代生物医学进展

Background & Objective:Background &Objective: The class Ⅰ Alcohol Dehydrogenases (ADH) play a key role in hepatic alcohol catabolism. Human ADH is encoded by at least seven genes, and three class Ⅰ ADH genes-ADH1, ADH2 and ADH3, which encode the α, β, and γ subunit respectively, had been isolated and mapped on chromosome 4q21-q25. This experiment tends to clone the human class Ⅰ ADH and investigate its role in the hepatic alcohol catabolism. Methods: A pair of primers were designed and the full-length cDNAs encoding human Class Ⅰ ADH were cloned at one time. Class Ⅰ ADH cDNAs were amplified with RT-PCR from total RNA extracted from fetal human liver and kidney, and cloned into pGEM-T vector. To identify cDNA segments, a pair of differential primers was designed. By using them, a portion of the ADHs which encodes the segment from -4 to 296 was cloned. These cDNA segments then were detected directly when being digested with Kpn Ⅰ and Pst Ⅰ, respectively. Then all the full-length cDNAs were subcloned in the plasmid pTYB11 and expressed in E. Coli. Stably. Alcohol Dehydrogenase activity of catalyzing alcohol were monitored at 340 nm. Results: Here we had successfully the human class Ⅰ ADH cloned and the full-length cDNAs expressed in E.col.I stably. The relative activity of recombinant enzymes metabolizing ethanol was 0.81 ～1.31 U/mg,0.09 ～0.15 U/mg and 0.76～1.11 U/mg, respectively. Conclusions: In the paper, the full-length cDNAs encoding human class Ⅰ AD H were successfully cloned and expressed and the recombinant enzymes showed the activities similar to the ones isolated from liver.

Effects of Panax notoginseng on the transdifferentiation of fibroblasts in human hypertrophic scar in vitro / 中华烧伤杂志

<p><b>OBJECTIVE</b>To observe the effects of Panax notoginseng on the transdifferentiation of the cultured human fibroblasts from hypertrophic scar in vitro, and explore its anti-fibrosis mechanism.</p><p><b>METHODS</b>The fibroblasts from human hypertrophic scar were cultured in vitro. Different amount of Panax notoginseng was added into the medium, respectively (400 microg/ml and 800 microg/ml). A culture without addition of the drug served as control. The fibroblast-populated collagen lattice method was used to detect the gel contraction, and contraction ratio was calculated. The immunocytochemistry staining method was used to detect the expression of alpha-smooth muscle actin. The flow cytometry method was used to detect the positive rate of alpha-smooth muscle actin.</p><p><b>RESULTS</b>The contraction degree of the fibroblasts after PNS administration was ameliorated at each time-point, with contraction index lower than that of controls (P < 0.05 or P < 0.01). Scattered distribution of alpha-SMA positive granules were observed in the cytoplasma, and the positive rate of alpha-SMA expression in 400 microg/ml (31.52%) and 800 microg/ml (24.28%) PNS groups were obviously lower than that in control group (45.74%, P < 0.05). The staining intensity of positive cells in 400 microg/ml and 800 microg/ml PNS groups was also obviously lower than that in control group (P < 0.05 or P < 0.01).</p><p><b>CONCLUSION</b>Panax notoginseng can inhibit the transdifferentiation of the cultured human fibroblasts from hypertrophic scar, and it exhibits an anti-fibrosis effect on hypertrophic scar in vitro.</p>

Cloning and Expression of the Full-length cDNAs Encoding Human Class Ⅰ Alcohol Dehydrogenases / 现代生物医学进展

Background & Objective:Background &Objective: The class Ⅰ Alcohol Dehydrogenases (ADH) play a key role in hepatic alcohol catabolism. Human ADH is encoded by at least seven genes, and three class Ⅰ ADH genes-ADH1, ADH2 and ADH3, which encode the α, β, and γ subunit respectively, had been isolated and mapped on chromosome 4q21-q25. This experiment tends to clone the human class Ⅰ ADH and investigate its role in the hepatic alcohol catabolism. Methods: A pair of primers were designed and the full-length cDNAs encoding human Class Ⅰ ADH were cloned at one time. Class Ⅰ ADH cDNAs were amplified with RT-PCR from total RNA extracted from fetal human liver and kidney, and cloned into pGEM-T vector. To identify cDNA segments, a pair of differential primers was designed. By using them, a portion of the ADHs which encodes the segment from -4 to 296 was cloned. These cDNA segments then were detected directly when being digested with Kpn Ⅰ and Pst Ⅰ, respectively. Then all the full-length cDNAs were subcloned in the plasmid pTYB11 and expressed in E. Coli. Stably. Alcohol Dehydrogenase activity of catalyzing alcohol were monitored at 340 nm. Results: Here we had successfully the human class Ⅰ ADH cloned and the full-length cDNAs expressed in E.col.I stably. The relative activity of recombinant enzymes metabolizing ethanol was 0.81 ～1.31 U/mg,0.09 ～0.15 U/mg and 0.76～1.11 U/mg, respectively. Conclusions: In the paper, the full-length cDNAs encoding human class Ⅰ AD H were successfully cloned and expressed and the recombinant enzymes showed the activities similar to the ones isolated from liver.