BACKGROUND:Fewer ethical issues exist in adult stem cells, and some operating technologies are relatively mature. Therefore, to construct tissue-engineered salivary glands using adult stem cells is very attractive and seductive with an extremely important application prospect. OBJECTIVE:To establish a rat model of salivary gland injury by ligation of the main excretory duct of the submandibular gland and to explore the existing feasibility and location of adult stem cells in the injured models. METHODS:The main excretory duct of the right submandibular glands was ligated in Sprague-Dawley rats. After 1 week, rats were kil ed to remove the bilateral glands that were then subject to hematoxylin-eosin staining, PAS glycogen staining and immunohistochemical staining for determination of CK-19, Bcl-2, Ki-67. After that, we compared the normal submandibular gland with the damaged model after ligation of main excretory duct. RESULTS AND CONCLUSION:The rats showed differences in the volume and mass of the affected and normal submandibular glands. The normal submandibular gland was oval, ruddy, smooth, soft with an intact envelop. After ligation, the injured submandibular gland appeared to have atrophy with dark red in color, irregular morphology, envelop congestion, and rough texture;the surrounding vessels showed compensatory expansion. PAS-positive gland cells disappeared, CK-19-postive smal duct epithelial cells proliferated, and laminin-positive cells that were rarely found in the normal gland existed around the duct. In addition, Bcl-2/Ki-67 positive cells were both increased. These findings indicate that stem/progenitor cells may be located in the periductal area of the submandibular gland;and the model of submandibular gland injury established by ligation of the main excretory duct is effective to activate stem/progenitor cells in the submandibular gland.

Objective:To explore the mechanism of gentiopicroside （GPS） in preventing acute liver injury induced by carbon tetrachloride （CCl₄） in mice and its effect on the Toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear factor-κB （NF-κB） signaling pathway. Method:Sixty mice were randomly divided into a normal control group， a model group， a silymarin group （150 mg·kg^-1）， and high- （200 mg·kg^-1）， medium- （100 mg·kg^-1）， and low-dose （50 mg·kg^-1） GPS groups， with 10 in each group. The mice in the groups with drug intervention were administered correspondingly by gavage at 10 mL·kg^-1， and those in the normal control group and the model group receive an equal volume of distilled water， once per day. Ten days after administration， mice in the normal control group were subjected to the intraperitoneal injection of peanut oil （10 mL·kg^-1） and those in other groups were injected with peanut oil （10 mL·kg^-1） containing 0.12% CCl₄ for the induction of acute liver injury model. After fasting for 16 hours， blood was collected from eyeballs and liver tissues were collected. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of liver tissues. The content or activities of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， alkaline phosphatase （ALP）， total superoxide dismutase （T-SOD）， and γ-glutamyl transpeptidase （γ-GT） in the serum， malondialdehyde （MDA） and glutathione peroxidase （GSH-Px） in liver tissues were determined by biochemistry techniques. The levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in liver tissues were measured by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the protein expression of TLR4， MyD88， and NF-κB in liver tissues. The expression of phosphorylated NF-κB （p-NF-κB） was detected by immunohistochemistry. Result:Compared with the normal control group， the model group showed increased levels of ALT， AST， ALP， TBIL， γ-GT， and MDA （P<0.01）， and blunted activities of T-SOD and GSH-Px （P<0.01）. Compared with the model group， the high- and medium-dose GPS groups exhibited declining levels of ALT， AST， ALP， TBIL， γ-GT， and MDA （P<0.05， P<0.01） and potentiated T-SOD and GSH-Px activities （P<0.05， P<0.01）. Compared with the normal control group， the model group displayed elevated levels of TNF-α， IL-1β， and IL-6 in liver tissues （P<0.01） and increased protein expression of TLR4， MyD88， and p-NF-κB （P<0.01）. Compared with the model group， the high- and medium-dose GPS groups showed decreased TNF-α， IL-1β， and IL-6 content in liver tissues （P<0.05， P<0.01） and dwindled TLR4， MyD88， and p-NF-κB protein expression （P<0.05， P<0.01）. Conclusion:GPS possesses a protective effect on mice with acute liver injury induced by CCl₄， and its mechanism of action may be related to the regulation of TLR4/MyD88/NF-κB signaling pathway and inhibition of oxidative stress.

Objective: To explore the protective effect of formula of Gougancai decoction (FGD) on acute liver injury induced by carbon tetrachloride (CCl₄) in rats, in order to provide basis for the development of pharmaceutical preparations or healthcare products. Method: Sixty rats were randomly divided into normal group, Silymarin group (120 mg·kg^-1) and FGD groups (475, 950, 1 900 mg·kg^-1). The normal group and the model group were given equal volume of saline by gavage, while the other groups were administered with the corresponding dose of drugs according to the body weight. After 10 days, the acute liver injury model was established with 12% carbon tetrachloride peanut oil solution (5 mL·kg^-1), except the normal group. All of the rats were put to death to collect serum and liver tissues. The contents of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), total bilirubin (TBIL), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected by biochemical methods, the levels of tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) in liver tissues were determined by enzyme-linked immunosorbnent assay(ELISA). Nuclear factor-κB (NF-κB) and peroxisome proliferator-activated receptor-γ (PPAR-γ) protein expression in liver tissues were detected by Western blot, and htoxylin eosin (HE) staining was used to observe the variation of liver histopathological. Result: Compared with the normal group, the serum activities of AST, ALT, ALP and the content of TBIL, MDA in the model group were significantly increased (P<0.01), the levels of TNF-α, IL-1β, IL-6 in liver tissue were remarkably increased (P<0.01), but the serum activities of SOD, GSH-Px were significantly decreased (P<0.01), the expression of NF-κB was enhanced in liver tissue (P<0.01), and PPAR-γ was down-regulated (P<0.01), indicating the successful modeling of acute liver injury. Compared with the model group, FGD could reduce the activities of AST, ALT, ALP and the contents of TBIL, MDA (P<0.05, P<0.01), decease the level of TNF-α, IL-1β, IL-6 (P<0.05, P<0.01), and down-regulate the expression of NF-κB (P<0.05, P<0.01), but up-regulate the activities of SOD, GSH-Px and the expression of PPAR-γ (P<0.05, P<0.01). The liver tissue lesions were alleviated to varying degrees. Conclusion: FGD has a protective effect on CCl₄-induced acute liver injury in rats, and its mechanism may be related to the activation of PPAR-γ and the inhibition of NF-κB signaling pathway, with anti-inflammatory and anti-oxidative effects.

Objective To explore the technique outcome of fixing the ventral stress pedicle screws into the injured vertebrae, as a method to enhance the posterior internal fixation. Methods From March 2002 to March 2005, 33 single thoracolumbar fractures were studied retrospectively. Among which, 16 cases were treated with the above method(group A), and the other 17 were treated with traditional two-level fixation(group B). Group A involved 11 males and 5 females, aged 48 years averagely(range, 32-74 years); and group B included 12 males and 5 females, aged from 21 to 61 years(mean, 40 years). All the patients underwent the operation within up to 3 weeks after fracture. For the injured vertebral bodies, their pedicles were intact on either unilateral or bilateral side, and their lower half and endplate were free from split. In group A, the pedicle screws in the injured vertebrae were used to achieve the ventral press vertical to the distraction for the stress neutralization, and also with the routine distraction and lordosis restoration, simultaneously. The mean follow-up period was 11 months with a range from 6 to 24 months. Results After operation, the optimal Cobb angle and anterior column restoration were achieved through the ventral reduction from the injured vertebral body, which was the contribution from the vertical stress pedicle screw. The degree in anterior movement of injured vertebrae pre- and postoperatively was much more in group A than group B, and the difference was of statistical significance(P

OBJECTIVE: To observe the effect and mechanisms of Bauhinia championii flavones(BCF) on hypoxia/reoxygenation (H/R) myocardial cells of neoanatal rats. METHODS: The cardiomyocytes hypoxia/reoxygenation injury model was developed, and Bauhinia championii flavones was pretreated (final concentration as 50, 100, 200 mg·L-1 respectively). The morphological changes of cardiomyocytes were observed by invert microscope. ELISA was used to evaluate the activities of tumor necrosis factor alpha, the protein expression of Bcl-2, Bax and NF-κβ was observed by immunohisto-chemistry, and Annex v-FITC/PI staining was used to detect apoptosis rate. RESULTS: Compared with model group, Bauhinia championii flavones pretreatment eased cardiomyocytes injury, decreased the activities of TNF-α and iNO S, maintained the eNOS level, up-regulated the expression of Bcl-2, down-regulated Bax and NF-κβ, and inhibited cadiocyte apoptosis (P<0.01 or P<0.05). CONCLUSION: BCF has protective effects on hypoxia/reoxygenation injury, which are associated with adjusting iNOS and NF-κβ signal channel, up-regulating Bcl-2, down-regulating Bax and NF-κβ, and inhibiting cadiocyte apoptosis.

Aim To investigate the protective effect of mogroside V on hydrogen peroxide ( H,02 )-induced oxidative stress response in mouse islet (3 cells MIN6 and the relation of its mechanism to PI3K/Akt signa¬ling pathway.Methods MIN6 cells were treated with 500 (jimol • L_1 H,(), after mogroside V,and cell via¬bility was detected by MTT.The release of reactive ox¬ygen species ( ROS) and apoptotic percentage of MIN6 cells were determined by flow cytometry.The expres¬sions of apoptosis-related factor Bel-2 , proliferation-re¬lated factor PCNA, protein Akt and p-Akt were deter¬mined by Western blot.Results H,02 restrained the proliferation of MIN6 cells obviously, induced ROS pro¬duction and apoptosis, and reduced the expression of Bel-2 and PCN A.The expressions of protein Akt and p-Akt decreased.After treatment of mogroside V , the release of ROS decreased, and the apoptosis of MIN6 cells was inhibited.The expression levels of apoptosis- related protein Bcl-2 and proliferation-related protein PCN A were reversed.The expressions of protein Akt and P-Akt increased.The viability of MIN6 cells in¬duced by H,0, increased.In addition, mogroside V partly reversed the apoptosis induction and ROS pro¬duction of Akt inhibitor MK2206 (5 jjimol • L"1 ) on MIN6 cells.Conclusions Mogroside V has protec¬tive effect on H202-induced oxidative damage in MIN6 cells and its mechanism is related to PI3K/Akt signa¬ling pathway.

OBJECTIVETo investigate the effect of Jianpi Yiqi Qingyou decoction on lymphocyte subsets and IL-2 mRNA in gastric tissue in rats with chronic superficial gastritis.

METHODWistar rats were randomly divided into 6 groups (11 for each): a blank control groups, the model of the control groups, the treatment groups (low-dose groups of traditional Chinese medicine, moderate-dose groups of traditional Chinese medicine, high-dose groups of traditional Chinese medicine) and lansoprazole groups. The models were made with the method in reference except a blank control groups. These rats are drinking freely with 0. 02% ammonia, continuous 90 days, and made preparations for experimental animal model of superficial gastritis. Making the model were detected by HE dying. The count of CD3+, CD4+ and CD8+ T cells were detected by immunohistochemistry. Using reverse transcriptase polymerase chain reaction (RT-PCR), the expression levels of IL-2 mRNA in gastric tissue were quantified.

RESULTCompared with that in model groups, the content of CD3+ T cells and CD4+ T cells in gastric tissue obviously increased in high dose of traditional Chinese medicine groups , the content of CD8+ T cells in gastric tissue obviously decreased in high dose of traditional Chinese medicine groups and the difference was significant (P < 0.01). The expression levels of IL-2 mRNA in gastric tissue obviously increased in moderate and high doses of traditional Chinese medicine groups, and the difference was significant compared with that in model group (P < 0.01).

CONCLUSIONJianpi Yiqi Qingyou decoction can obviously improve the content of CD3+ T cells and CD4+ T cells and the expression levels of IL-2 mRNA, decrease the content of CD8+ T cells in gastric tissue, improve immunity of rats. So the research results can provide some evidences for the treatment for chronic superficial gastritis.

Animals ; CD4-CD8 Ratio ; Chronic Disease ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gastritis ; genetics ; immunology ; pathology ; Interleukin-2 ; genetics ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach ; drug effects ; immunology ; metabolism ; pathology ; T-Lymphocyte Subsets ; drug effects ; metabolism

OBJECTIVE： To study the in vitro inhibitory effects and antioxidant activity of different solvent extracts of Boehmeria nivea leaves against influenza A virus（H1N1）， and to expand the medicinal parts of B. nivea and develop natural antiviral and antioxidant drugs. METHODS： The leaves of B. nivea were extracted with 95％ ethanol. The ethanol extract was dissolved by water heating， and extracted with different solvents to obtain petroleum ether phase， trichloromethane phase， ethyl acetate phase， n-butanol phase and aqueous phase extracts of B. nivea leaves. The toxicity of aqueous extract of B. nivea leaves （50-400 μg/mL） on Madin-Darby canine kidney （MDCK） cell line was investigated. Using ribavirin as positive control， MDCK cells were attacked by influenza A virus（H1N1）. Western blotting assay was used to detect the expression of nucleoproteins （NP） in viral infected cells after treated with same concentrations of petroleum ether phase， trichloromethane phase， ethyl acetate phase， n-butanol phase and aqueous phase extracts of B. nivea leaves （100 μg/mL）， different concentrations of aqueous phase extract solution of B. nivea leaves （50， 100， 200， 400 μg/mL） and different concentrations of ribavirin solution （0.31， 0.63， 1.25 μg/mL）. Using vitamin C as a positive control， hydroxyl radical（·OH） scavenging test， DPPH radical scavenging test and reduction test were used to investigate in vitro antioxidant activity of the extracts. RESULTS： Aqueous phase extract of B. nivea leaves with concentration less than 400 μg/mL was nontoxic to MDCK cells. The petroleum ether phase， trichloromethane phase， ethyl acetate phase and aqueous phase extracts at 100 g/mL could significantly reduce the expression of NP protein in influenza A virus（H1N1） infected cells （P＜0.01）. Different concentrations （50-400 μg/mL） of aqueous extract could significantly reduce the protein expression of NP （P＜0.01） in concentration-dependent manner. The in vitro antioxidant activity of petroleum ether phase and ethyl acetate phase was similar to that of vitamin C. CONCLUSIONS： B. nivea leaves extract have better anti-influenza A virus（H1N1） effects in vitro， and the extracts of petroleum ether phase and ethyl acetate phase show good antioxidant activity in vitro.

Objective PINK1 and Parkin are directly invoveled in the regulation and maintenance of mitochondrial functional morphology. We aim to explore the effect of Wnt2 overexpression on PINK1^B9 Mutant Drosophila and its mechanism in this study. Methods The GAL4-UAS system was used to construct the normal control flies（W1118/ + ；MHC-GAL4/+）, PINK1^B9 transgenic Drosophila model flies（UAS-PINK1^B9 /y；MHC-GAL4 / +；Parkinson's disease model of Drosophila melanogaster), the Wnt2 overexpression flies（UAS-PINK1^B9 /y；MHC-GAL4 / Wnt2 OE） and the Wnt2 RNAi flies（UAS-PINK1^B9 /y；MHC-GAL4 /Wnt2）. On the 5th day, the abnormal wings phenotype rate and flying rate of flies were observed. The contents of Ndufs3 proteins were detected by Western blot. The mRNA expression levels of PGC-1α, Nrf1 and TFAM related to mitochondrial metabolism and synthesis were detected by real-time fluorescence quantitative PCR. The morphology of mitochondria was observed by electron microscopy. Complex I and Complex II function was detected by high-resolution mitochondrial respiratory system. Results Compared with the normal control flies, PINK1^B9 transgenic Drosophila model flies showed increased abnormal wings phenotype rate([1.87±0.06]% vs [68.79±0.70]%), decreased flying rate([ 97.51±0.52)% vs(3.95±0.53)%], and the differences were statistically significant(P<0.05); Compared with PINK1^B9 transgenic Drosophila model flies, the Wnt2 RNAi flies showed decreased abnormal wings phenotype rate[(10.14±1.72)%], increased flying rate([41.83±2.57]%)(P<0.05). Compared with the normal control flies, PINK1^B9 transgenic Drosophila model flies showed decreased expression levels of PGC-1α and Nrf1,Ndufs3 proteins, Complex I and Complex II(P<0.05);On the contrary, the Wnt2 RNAi flies showed increased trends compared with PINK1^B9 transgenic Drosophila model flies(P<0.05). Conclusion Overexpression of Wnt2 protects PINK1^B9 transgenic Drosophila models, which is related to the improvement of mitochondrial function.

OBJECTIVE: To investigate the effect of ANP32A silencing on invasion and migration of colon cancer cells and the influence of the activity of AKT signaling pathway on this effect. METHODS: Colorectal cancer HCT116 and SW480 were transfected with a small interfering RNA targeting ANP32A via a lentiviral vector. At 24, 48 and 72 h after the transfection, the changes in cell proliferation and AKT activity in the cells were detected using MTT assay and Western blotting, respectively. HCT116 and SW480 cells were treated with the AKT agonist SC79 or its inhibitor MK2206 for 24, 48, 72 and 96 h, and the changes in cell migration and invasion ability were analyzed using Transwell chamber assay and cell proliferation was assessed using MTT assay. The effects of SC79 and MK2206 on migration and invasion abilities of HCT116 and SW480 cells with or without ANP32A silencing were examined using wound healing and Transwell chamber assays, and the changes in the expression of metadherin (MTDH), a factor associated with cells invasion and migration, was detected with Western blotting. RESULTS: Lentivirus-mediated ANP32A silencing significantly down-regulated the activity of AKT and inhibited the proliferation of both HCT116 and SW480 cells (P < 0.01). The application of AKT inhibitor MK2206 obviously inhibited the proliferation, invasion and migration of the colorectal cancer cells (P < 0.05), while the AKT agonist SC79 significantly promoted the invasion and migration of the cells (P < 0.01). In HCT116 and SW480 cells with ANP32A silencing, treatment with MK2206 strongly enhanced the inhibitory effects of ANP32A silencing on cell invasion and migration (P < 0.05) and the expression of MTDH, while SC79 partially reversed these inhibitory effects (P < 0.01). CONCLUSION ANP32A silencing inhibits invasion and migration of colorectal cancer cells possibly by inhibiting the activation of the AKT signaling pathway.
Humans ; Proto-Oncogene Proteins c-akt ; Cell Proliferation ; Blotting, Western ; Cell Movement ; Colonic Neoplasms ; Membrane Proteins ; RNA-Binding Proteins/genetics* ; Nuclear Proteins