[Effects of methyl cantharidimide tablets on urinary protein and enzymes in Beagle dogs].

OBJECTIVE: To investigate the nephrotoxic effects of methyl cantharidimide tablets on urinary protein and enzymes in Beagle dogs. METHOD: Beagle dogs were randomly divided into negative control group(blank tablet), methyl cantharidimide tablets group (6.11,12.21, 24.42 mg x kg(-1)), continuously 30 days of oral adminiStration, once a day. The drug and control group were collected and determined fresh urine in 1, 2, 3 and 4 weeks of the administration; Serum urea nitrogen (BUN), creatinine (Crea), total protein (TP) and albumin (ALB) as well as sodium, potassium, chloride electrolyte were determined on 15 and 30 days of the administration; Urine albumin (mAlb), kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin( NGAL), N-acetyl-beta-D-glucosaminidase (NAG), clusterin, beta2-microglobulin (beta2-MG), alpha1-microglobulin (alpha1-MG), alanine aminopeptidase( AAP) and im- munoglobulins IgG were tested on 15 and 30 days of the administration. RESULT: Compared with the control group, urine protein and white blood cells was significantly increased in each dose group. On 15 days of the administration, mAlb were higher in each dose group, KIM-1, NGAL, clusterin, NAG and AAP were significantly higher in high-dose group, while the middle and low dose group had no significant difference, as well as blood SCr and BUN no obvious abnormalities. On 30 days, mAlb, KIM-1, clusterin, NAG, AAP were increased in each dose group, appearing dose-effect relationship, beta2-MG and NGAL levels were significantly increased in high-dose group. Contents above indicators were increased with significant dose and time relationship, and serum BUN, Scr were correlated, suggesting that urine mAlb, KIM-1, clusterin, NAG and AAP indicators that can sensitively respond the changes of proteins and enzymes in urine. CONCLUSION: Methyl cantharidimide tablets has a renal toxicity, urine mAlb, KIM-1, clusterin, NAG and AAP can be used as the early nephrotoxic biomarkers of methyl cantharidimide tablets.

Equivalence of the Wei potential model and Tietz potential model for diatomic molecules.

By employing the dissociation energy and the equilibrium bond length for a diatomic molecule as explicit parameters, we generate improved expressions for the well-known Rosen-Morse, Manning-Rosen, Tietz, and Frost-Musulin potential energy functions. It is found that the well-known Tietz potential function that is conventionally defined in terms of five parameters [T. Tietz, J. Chem. Phys. 38, 3036 (1963)] actually only has four independent parameters. It is shown exactly that the Wei [Phys. Rev. A 42, 2524 (1990)] and the well-known Tietz potential functions are the same solvable empirical function. When the parameter h in the Tietz potential function has the values 0, +1, and -1, the Tietz potential becomes the standard Morse, Rosen-Morse, and Manning-Rosen potentials, respectively.

[Function of STAT1 in proliferation and differentiation of human hypertrophic scar fibroblast induced by connective tissue growth factor].

OBJECTIVE: To validate whether STAT1 participated in the process of CTGF-induced proliferation and differentiation of human hypertrophic scar fibroblast (hHSF) on account of past research. METHODS: To cultivate hHSF with 6 patients' hypertrophic scar specimens together. Electrophoretic mobility shift assay (EMSA) was used to verify binding ability between DNA and STAT1 with the stimulus of different concentration CTGF (0, 5, 7.5, 10, 15 ng/ml) at 45th min and the stimulus of 10 ng/ml CTGF at different phase point (0, 10, 20, 30, 45, 60, 90, and 120 min). We divided cells into CTGF group, STAT1 ASODN group. STAT1 ASODN + CTGF group. control group. And MTT was used to detect the proliferation of hHSF on days1, 2 and 3, and RT-PCR to detect alpha-smooth muscle actin mRNA to follow the differentiation. RESULTS: EMSA showed that the binding ability between STAT1 and DNA depended on the concentration of CTGF and peaked with the stimulation of 10ng/ml CTGF. And at the same time, it peaked at 45 - 60 min with 10 ng/ml CTGF. MTT showed that cell proliferation of CTGF group was much higher than that of control group (all P < 0.05). And those of STAT1 ASODN group and STAT1 ASODN + CTGF group were much lower than those of control group and CTGF group (all P < 0.05). RT-PCR showed that differentiation activation from fibroblast to myofibroblast of CTGF group, STAT1 ASODN group, STAT1 ASODN + CTGF group and control group were 0.78 +/- 0.08, 0.38 +/- 0.09, 0.76 +/- 0.10, and 0.40 +/- 0.12, respectively. Differentiation activation of STAT1 ASODN group and control group were much lower than those of CTGF group and STAT1 ASODN + CTGF group (all P < 0.05). CONCLUSION: STAT1 ASODN is important in the process of proliferation of hHSF and it blocks the stimulation of CTGF on hHSF proliferation. The above result revealed that STAT1 participates in the process of hHSF proliferation induced by connective tissue growth factor.

Binding interaction of sodium benzoate food additive with bovine serum albumin: multi-spectroscopy and molecular docking studies.

Sodium benzoate (SB) is widely used as a preservative in food industry, and bovine serum albumin (BSA) is a major carrier protein similar to human serum albumin (HSA), the study of the binding between the two has great significance on human health. In this paper, we systematically investigated the binding of SB and BSA under the simulated physiological conditions combining with various common analytical methods, e.g., fluorescence, UV-vis absorption, synchronous fluorescence and circular dichroism (CD) spectra, as well as molecular docking method. The fluorescence quenching measurements were respectively carried out at 298 K, 303 K and 308 K using the Stern-Volmer method. The results reveal that ground state SB-BSA complex was formed within the binding constants from 2.02 × 104 to 7.9 × 103 M-1. Meanwhile, the negative values of ΔH 0 (- 43.92 kJ mol-1) and ΔS 0 (- 111.6 J mol-1 K-1) demonstrated that both the hydrogen binding interaction and van der Waals forces contributed to stabilizing the SB-BSA complex. The site marker competitive experiments show that the SB and BSA bound at site I. Furthermore, the experimental results of UV-vis absorption, synchronous fluorescence and CD spectra indicate that the binding of SB and BSA may change the conformation of BSA. In addition, the molecular docking experiment suggests that hydrogen bond was formed in the interaction between SB and BSA.

Thermodynamic Properties for Carbon Dioxide.

We first report three reliable analytical expressions of the entropy, enthalpy and Gibbs free energy of carbon dioxide (CO2) and perform predictions of these three thermodynamic quantities on the basis of the proposed analytical expressions and in terms of experimental values of five molecular constants for CO2. The average relative deviations of the calculated values from the National Institute of Standards and Technology database over the temperature range from 300 to 6000 K are merely 0.053, 0.95, and 0.070%, respectively, for the entropy, enthalpy, and Gibbs free energy. The present predictive expressions are away from the utilization of plenty of experimental spectroscopy data and are applicable to treat CO2 capture and storage processes.

Effect of Danggui Buxue Tang on immune-mediated aplastic anemia bone marrow proliferation mice.

To investigate the pharmacological effects of Danggui Buxue Tang (DBT) on immune-mediated aplasia anemia mice. The model of immune-mediated aplasia anemia mice was induced by means of (60)Co γ-ray irradiation and mixed cells of thymus and lymphnode of DBA/2 mice infusion through tail vein, the parameters tested indices were as following: blood picture, bone marrow nucleated cell count (BMNC), murine colony-forming unit-megakaryocytes (CFU-GM) of bone marrow cells, murine colony-forming unit-erthroid (CFU-E) and burst forming unit-erythroid (BFU-E). The results showed that DBT could not only withstand significantly decreation of blood cells by immune-mediated, but also stimulate on the growth of bone marrow colony cell and increase the weight of hemopoietic progenitor of bone marrow. Therefore, DBT had an obvious treat effect on immune-mediated aplasia anemia models mice.

[The cellular and molecular mechanism of inhibitory effect of asiaticoside on capsular contracture following breast augmentation].

OBJECTIVE: To explore the cellular and molecular mechanism of the inhibitory effect of asiaticoside on capsular contracture following breast augmentation. METHODS: 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. RESULTS: 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). CONCLUSIONS: 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.

[Genetic characteristic of wild-type measles viruses isolated in Ningbo from 2004 to 2008].

OBJECTIVE: To investigate the genetic characterizations and genotypes of measles viruses that was prevalent in Ningbo city. METHODS: Measles viruses were isolated from throat swab specimens of suspected measles cases from 2004 to 2008 and 456 bp fragments of C terminus of nucleprotein (N) gene were amplified by RT-PCR. Sequence analysis was conducted of all 22 virus strains while compared with other measles virus strain which published in GenBank. RESULTS: All of the 22 measles isolates belonged to genotype H1. The homogeneities of 456 bp fragments of C terminus of nucleprotein (N) gene were as high as 97.1%-98.5%,98.2%-99.6% and 98.5%-98.9% respectively. The nucleotide (amino acids) variability was 8.0%-9.5% (9.8%-14.5%) compared to S191. CONCLUSION: Genotype H1 measles virus circulated in Ningbo, China from 2004 to 2008. H1a was the predominant epidemic strain, H1b strain was existed as well.

[To transdifferentiate human hypertrophic scar fibroblasts induced by connective tissue growth factor mediated transforming growth factor-beta 1 in vitro].

OBJECTIVE: To investigate the role of connective tissue growth factor (CTGF) induced TGF-beta 1 in the transdifferentiation of human hypertrophic scar fibroblast (HSFb). METHODS: 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. RESULTS: 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. CONCLUSIONS: CTGF is one of the most important downstream efforts for TGF-beta 1 in inducing the transdifferentiation of HSFb.

[Modality and structure of Japanese eel (Anguilla japonica) during spermiogenesis].

The special process and special structure which bring organelle during the spermiogenesis of Japanese eel, Anguilla japonica were observed by scanning electron microscope and transmission electron microscope. The process which spermatoblast became sperm including four special stages, the early stage, the middle stage, the later stage and the spermic stage, then came into being a normal mature sperm. In the early stage, cell nucleus became long form gradually by the oval form. In one side of the cell nucleus, there was a big and special globoid structure dyeing lower, account for 1/3 of cell nucleus cubage. It contains a little of deep dyeing grain form and the lines form material, the outside is wrapped by plasmalemma separated with the cell nucleus, the outside of that structure and cell nucleus still lay a plasmalemma. The spermiogenesis of early stage did not form independent centriolar complex and mitochondria. In the middle stage, the cell nucleus presented a long form with the globoid structure on the top of the nucleus, and the down side had no globoid structure where the flagellum primordium appears. The globoid structure changed with the spermiogenesis. The inner part of the globoid differentiated a centriolar complex and mitochondria step by step. The lysosomes distributed in the medium segment of the cell nucleus obviously. In the late stage, the cell nucleus was similar with the shape of eyebrow or crescent. The centriolar complex released from the globoid structure, then became an independent structure. There were mitochondria which had not become the independent structure still in the globoid structure. Under the karyon, there was flagellum primordium where sent a rather long flagellum. The flagellum formed a typical "9+0" microtubular structure at that time. The spermatozoa in this phase has movable ability. In the spermic stage, the cell nucleus was round in shape. The centriolar complex was inside implantation fossa. Mitochondria were under karyon. And under the mitochondria was the central space of the sleeve. The flagellum formed a typical "9+2" microtubular structure at that time. The spermatozoa of Japanese eel, A. japonica became complete mature spermatozoa must pass through four phases for abnormality.

[Reproductive biology and artificial propagation of Acipenser sinensis below Gezhouba Dam].

A total of 36 females and 21 males of Chinese sturgeon Acipenser sinensis were caught in 1998-2004 excluding 2002 to study the characteristics of their reproductive biology and the effect of their artificial propagation. The results showed that the body length (BL), body mass (BM) and age of the females were 240-320 cm, 140-432 kg, and 15-30 years, and those of the males were 153-284 cm, 70-244 kg and 12-26 years, respectively. The inducing rate was 93.1% for females and 100% for males, and the ova had 7 different colors. The absolute fecundity was 200,000-590,000 eggs, with an average of 358,000 eggs, and the relative fecundity to BM was 820-3,020 eggs per kg, with an average of 1,590 eggs per kg. The sperm had 4 different colors. The absolute sperm quantity obtained from one male was 1,000-5,952 ml, with an average of 2,597.8 ml, and the relative sperm quantity to BM was 1.25-31.24 ml . kg(-1), with an average of 13.3 ml . kg(-1). During the study period, the average fertilization rate in artificial propagation was 63.7%, and the hatching rate was 48.1%, with 4,762,000 fry obtained. Compared with the data in 1976, the natural reproductive capacity of the Chinese sturgeon broodstocks declined greatly.

[Effects of Panax notoginseng on the transdifferentiation of fibroblasts in human hypertrophic scar in vitro].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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). CONCLUSION: 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.

[Expression of connective tissue growth factor gene in the hypertrophic scar and keloid tissue].

OBJECTIVE: To explore the effect of connective tissue growth factor on the pathogenesis of hypertrophic scar and keloid tissue. METHODS: The content of hydroxyproline was determined and the expression of connective tissue growth factor gene was detected by the reverse transcription-polymerase chain reaction and image analysis technique in 5 normal skins, 15 hypertrophic scars and 7 keloid tissues. RESULTS: The contents of hydroxyproline in the hypertrophic scar (84.10 +/- 1.76) and keloid tissue (92.38 +/- 2.04) were significantly higher than that of normal skin tissue (26.52 +/- 4.10) (P < 0.01). The index of connective tissue growth factor mRNA in the hypertrophic scar (0.78 +/- 0.63) and keloid tissue (0.84 +/- 0.04) were higher than that of normal skin tissue (0.09 +/- 0.25) (P < 0.01). CONCLUSION: Connective tissue growth factor may play an important role in promoting the fibrotic process of hypertrophic scar and keloid tissue.

[Effects of antisense oligonucleotides on the expression of connective tissue growth factor gene and on the collagen synthesis in the cultured human keloid fibroblasts].

OBJECTIVE: To study the role of connective tissue growth factor (CTGF) in the pathogenesis of human keloid. METHODS: Human keloid fibroblasts (HKF) were isolated from human keloid and cultured in vitro. The cells were then divided into 3 groups according to different processing, i.e. ASODN treatment (AT), in which phosphorothioate CTGF antisense oligonucleotides (ASODN) labeled by fluorescent isothiocyananate were transfected into the HKFs by liposome; liposome control (LC, with liposome only); control groups (without liposome or ASODN). The distribution of CTGF ASODN in all groups of cells was observed under fluorescent microscope. The CTGF mRNA index (RI) of HKF was assessed by reverse transcription polymerase chain reaction method (RT-PCR). The collagen synthesis of HKF was assessed by (3)H-proline incorporation method. RESULTS: A large amount of fluorescence could be observed in the cytoplasm of HKFs in AT 12 hours after transfection, but not in LC and C groups. The CTGF mRNA index of HKF in AT group 48 hours after transfection was significantly lower than that in LC and C groups (0.12 +/- 0.62 vs 0.51 +/- 0.18 vs 0.54 +/- 0.35, P < 0.01). The (3)H-proline incorporation rate in AT group (108.96 +/- 79.05) was lower than that in LC and C groups (P < 0.01). CONCLUSION: The expression of CTGF gene and collagen synthesis of the cultured HKF could be inhibited by CTGF ASODN, implying that CTGF played a role in the development of excessive fibrosis of human keloid.

[Collagen synthesis and expression of connective tissue growth factor in the cultured fibroblasts of human hypertrophic scar].

OBJECTIVE: To explore the effect of connective tissue growth factor on the pathogenesis of human hypertrophic scar. METHODS: Normal skin and hypertrophic scar fibroblasts were cultured in vitro. The collagen synthesis of fibroblasts were measured by H3-proline incorporation method. The expression of connective tissue growth factor protein and mRNA of fibroblasts were detected with immunocytochemistry staining and reverse transcription polymerase chain reaction methods. RESULTS: Compared with normal skin fibroblast, the collagen synthesis and the expression of connective tissue growth factor protein and mRNA in the hypertrophic scar fibroblast was higher (P < 0.01). CONCLUSION: Connective tissue growth factor may play an important role in promoting the fibrotic process of hypertrophic scar.

[Effects of connective tissue growth factor antisense oligonucleotides on the cultured human keloid fibroblasts in vitro].

OBJECTIVE: To explore the effects of connective tissue growth factor (CTGF) on the pathogenesis of human keloid. METHODS: CTGF antisense oligonucleotides (ASODN) conjugated with isothiocyanate fluorescence was encapsulated by liposome, and then added into the human keloid fibroblast (HKF) culturing media. The intracellular distribution of CTGF ASODN was observed with fluorescence microscopy in the fixed HKF. The proliferation of HKF was measured by MIT test. The apoptosis of HKF was measured with a flow cytometer. The collagen synthesis of HKF was measured by using H3-proline incorporation method. RESULTS: The CTGF ASODN inhibited the proliferation and collagen synthesis of the HKF, compared with the control, but it increased the apoptosis after the transfection (P < 0.01). CONCLUSION: CTGF ASODN may has anti-fibrotic effects on human keloid in vitro, and the CTGF may play an important role in promoting the fibrosis of human keloid.

[Establishment of an animal model of human hyperplastic scar in nude mice].

OBJECTIVE: To establish an ideal model of human hyperplastic scar (HS) in nude mice, so as to provide us a new model to carry out further studies on the mechanism of HS development. METHODS: Full skin defect sized 2.0 cm x 1.5 cm was created on the back of 100 nude mice. The defect was thereafter covered with full thickness human skin. After the grafted skin survived, the nude mice were subjected to deep partial thickness burn of the grafted skin with heated copper rod. The development of the hyperplasia of the scar after wound healing was observed histologically and grossly. RESULTS: Grafted full-thickness human skin took and survived well in 86 out of 100 nude mice. There was obvious and continuous hyperplasia of scar in 67 mice (78%). The external appearance and histological features of the HS appeared similar to those in human HS. The average thickness of the scar was 0.34 cm, with the thickest part measuring 0.6 cm. In addition, the time of hyperplastic change lasted for 63 - 217 days in average of 128 days. CONCLUSION: Obvious and continuous scar hyperplasia could be found in this model, and the whole process beginning from wound healing to the formation of HS could be easily observed. The model was therefore suitable and ideal for the study of HS.