OBJECTIVETo observe the effect of Tangke Decoction (TD) on the expression of TGF-beta1/Smad4 of rats with early diabetes and to explore the effect and mechanism of TD against the renal injury induced by diabetes.

METHODSSD rats were randomly divided into the normal control group (n = 12), the model group (n = 10), the Chinese herbs prevented group (n =10), the Chinese herbs treated group (n = 10), and the Western medicine control group (n = 10). TD (18 mg/kg) was given by gastrogavage to rats in the Chinese herbs prevented group immediately after successful modeling for 12 weeks, once daily. At the 4th week of successful modeling, rats in the rest 4 groups were administered by gastrogavage. Equal volume of normal saline was given to rats in the model group and the normal control group. Benazepril suspension (1 mg/kg) was administered by gastrogavage to rats in the Western medicine control group for 8 weeks, once daily. TD (18 mg/kg) was given by gastrogavage to rats in the Chinese herbs treated group for 8 weeks, once daily. The body weight, kidney weight, index of kidney weight, fasting blood sugar, 24 h urinary albumin excretion rate were examined after experiment. The pathological changes of the renal tissue were observed by HE staining, Masson staining, and electron microscope. The expression of renal transforming growth factor-beta1, (TGF-beta1) and Smad4 were detected using immunohistochemical assay.

RESULTSCompared with the normal control group, the body weight of rats decreased significantly; the kidney weight, index of kidney weight, blood sugar, 24 h urinary protein excretion, the urinary albumin excretion rate,TGF-beta1 and Smad4 expression increased significantly in the model group (all P < 0.01). Compared with the model group, the aforesaid indices were improved in each treatment group with statistical difference (P < 0.05, P < 0.01). Compared with the Western medicine control group, the kidney weight, index of kidney weight, blood sugar, 24 h urinary protein excretion, and the urinary albumin excretion rate were obviously improved in the Chinese herbs prevented group (P < 0.01). The renal pathological changes were most obvious in the model group significantly, but they were improved in all treatment groups.

CONCLUSIONTD could obviously improve the symptoms of diabetes and down-regulate the expression of renal TGF-beta1 and Smad4 of early diabetic nephropathy rats, which suggested that TD had certain preventive effect on early diabetic nephropathy.

Animals ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Kidney ; metabolism ; Male ; Rats ; Rats, Wistar ; Smad4 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism

Objective To develop an online organ doses reporting software VirtualDose-IR, which can compute the radiation doses and provide an easy access to evaluation and control of patients ′radiation doses.Methods Monte Carlo method was applied to simulating various interventional radiology ( IR) processes , which included various parameters such as different patient models at different ages and with different weights , different projection angles and regions of interest , and other parameters .All of the dose data was acquired and then integrated into a database , and displayed with hyper text markup language (HTML), so only a web browser was necessary for users .Results A web-based software that reports organ doses for patients under IR progress was developed .The organ doses assessed with VirtualDose-IR were compared with other experiment and simulation data , and the results were basically consistent with each other .Conclusions VirtualDose-IR is a easy and efficient method to assess patients′radiation doses of IR.

Object: To investigate the effects of Huaiqihuang Granule, a compound Chinese herbal medicine, on expressions of nephrin and podocin of slit diaphragm of glomerular podocytes in rats with adriamycin-induced nephrosis and to explore the mechanism in reducing the proteinuria. Methods: Twenty SD rats were randomly divided into five groups: control group, model group, glucocorticoid group, Huaiqihuang Granule group and Huaiqihuang Granule plus glucocorticoid group. The 24-hour urine was collected 7, 14, 21 and 28 days after adriamycin injection respectively to measure 24-hour urinary protein, and all rats were sacrificed after 28-day treatment. Pathological changes in renal tissues were observed under a light microscope and an electron microscope. Expressions of nephrin and podocin mRNAs in renal cortex were determined by real-time polymerase chain reaction, and protein levels of nephrin and podocin were detected by Western blotting. Results: (1) In the model group and the treatment groups, the level of urinary protein increased significantly from the 14th day. (2) Under the light microscope, inflammatory cells and slight fibroplasia were found in renal interstitium of the model group, but there were less inflammatory cells in renal interstitium in the intervention groups than in the model group. Under the electron microscope, 29 days after adriamycin injection, extensive fusion of foot processes was observed. (3) The expressions of nephrin and podocin were higher in treatment groups than in the model group. (4) Proteinuria level was negatively correlated with the expressions of nephrin mRNA and nephrin and podocin proteins. Conclusion: The above results indicate that Huaiqihuang Granule can maintain the integrity of the slid diaphragram in podocyte, alleviate the lesion of glomerular filtration membrane, and decrease the proteinuria by up-regulating the expressions of nephrin and podocin. Huaiqihuang Granule plus glucocorticoid maybe has better effects than glucocorticoid alone.

Objective To evaluate the security of big animals with a new bioartificial liver system .Methods Six Tibet pigs re-spectively received treatment of hybrid artificial liver and simple bioartificial liver ,observed and recorded the vital signs ,venous pressure ,transmembrane pressure and slurry pot pressure each hour ,and collected blood to make endotoxin and bacterial culture test in the zero hour ,fourth and eighth hour .Results Compared with the zero hour ,venous pressure ,transmembrane pressure ,slur-ry pot pressure were much higher in the fifth hour (P< 0 .05) ,and there were no significant difference in the rest of other time points(P>0 .05) .The mean arterial pressure and respiratory rate in all time point showed no significant changes (P>0 .05) .Com-pared with the zero hour ,the heart rate was much lower in the second hour (P<0 .01) .The values of blood endotoxin were less than 0 .5 EU/mL in the zero hour ,fourth and eighth hour from beginning ,and the bacterial culture test showed no growth of bacteria . Conclusion The experiment of big animals with a new bioartificial liver system was safe ,the efficacy in the treatment of hepatic failure could be assessed further .

Objective To retrospectively analyze 45 cases of oral and maxillofacial malignant tumors and to assess the efficacy and feasibility of oral and maxillofacial malignant tumors implanted with 125I radioactive seeds under guidance of ultrasound.Methods A total of 47 focuses in these 45 patients were determined with the size of these tumors by imaging study,the section was planed by ultrasound,the number and distribution of radioactive seeds were determined with the help of the particle treatment planning system,and were percutaneously implanted particles under guidance of ultrasound.The number and the distribution of particles were assessed by CT.Efficacy endpoints were reexamined and evaluated regularly by ultrasonic and CT according to the standards of WHO.Results The total percentage of efficacy was 70.2% (including complete remission,partial remission).The treatment effect of metastatic carcinoma of lymph node is superior to the parotid tumor.There was no serious complication during the period of implanting and 2 patients with oral ulcers were found after operation.Conclusion The oral and maxillofacial malignant tumor treated implanted with 125I radioactive seeds under guidance of ultrasound is very effective and safe,which is deserved to popularize.The ultrasound is the first choice as a guided method for oral and maxillofacial malignant tumors.

OBJECTIVETo investigate whether hypertonic saline (HS) can induce the synthesis and release of glutamate in cultured hypothalamic astrocytes or C6 cell line.

METHODSAstrocytes were isolated, cultured, purified and identified from the hypothalamus of newborn rat (1 day). The astrocytes were randomly divided into five groups: isotonic (IS) and HS groups, astrocytes were incubated by IS and HS (320 mosM NaCl) medium, respectively, for 1, 3, 5, 10 or 15 min; carbenoxolone (CBX)+IS and CBX+HS groups, astrocytes were pre-treated with CBX (100 mmol/L) for 1 h at 37 degrees C in a 5% CO(2) / 95% atmosphere, then removed to IS and HS medium, respectively, for 1, 3, 5, 10 or 15 min; Ca(2+)+HS group, astrocytes were pre-incubated with Ca Ca(2+) (1,000 micromol/L) for 1 h at 37 degrees C in a 5% CO(2) / 95% atmosphere, followed by a wash with isotonic FBS/DMEM, and then removed to hypertonic saline for 1, 3, 5, 10 or 15 min. The media of five groups were collected to analyze the medium glutamate concentration with high performance liquid chromatography. The astrocytes were fixed and double immunofluorescent stained with anti-glial fibrillary acidic protein (GFAP) and anti-glutamate. The C6 cells were divided into four groups: IS, HS, CBX+IS and CBX+HS groups, and used for quantitative measurement of glutamate in cells by flow cytometry (FCM).

RESULTS(1) Anti-GFAP immunofluorescent signal revealed no significant difference among various time points in each group, or among the five groups. (2) The anti-glutamate immunofluorescent signal was increased in HS group and peaked at 5 min, and decreased and returned to the level of IS group at 15 min (P < 0.01 vs the 5 min of HS group). In CBX+HS group, the glutamate intensity was higher than that in CBX+IS and HS groups. (3) The medium glutamate concentration had no change after treatment with HS for 1 and 3 min, while increased markedly after treatment for 5 min to 15 min (P< 0.01 vs 1 min and 3 min). On the contrary, the medium glutamate concentrations in the CBX+HS or Ca(2+)+HS group were significant lower than that in the HS group (P < 0.01). (4) FCM showed HS and CBX+HS induced glutamate increase in C6 cells.

CONCLUSIONHS induced cultured rat hypothalamic astrocytes or C6 cells to synthesize and release glutamate; CBX could block glutamate release, but could not disrupt glutamate synthesis.

Analysis of Variance ; Animals ; Animals, Newborn ; Astrocytes ; drug effects ; metabolism ; Calcium ; pharmacology ; Carbenoxolone ; pharmacology ; Cells, Cultured ; Chromatography, High Pressure Liquid ; methods ; Flow Cytometry ; Gene Expression Regulation ; drug effects ; Glial Fibrillary Acidic Protein ; metabolism ; Glutamic Acid ; metabolism ; Hypothalamus ; cytology ; Rats ; Saline Solution, Hypertonic ; pharmacology ; Time Factors

OBJECTIVETo study DNA and nucleus damage in human embryo lung fibroblast (HLF) exposed to hydroquinone (HQ) and its genotoxicity.

METHODSHLF were treated with HQ (0, 10, 20, 40, 80 micro mol/L, respectively) for 3 h and DNA damage was detected by comet assay. HLF was also treated with the same concentrations of HQ for 1 h and micronucleus test was performed after they were cultured for 24 h.

RESULTSComet assay showed that percentage of cells with tails in each groups treated with varied doses of HQ was 12%, 19%, 42%, 79% and 95%, respectively, with mean tail length of 7.87, 9.35, 11.03, 19.28 and 23.32 micro m, respectively, in an obvious dose-dependent manner (P < 0.05). Very significant increase in percentage of cells with tails and length of their comet tail were observed in those groups treated with HQ of 20, 40 and 80 micro mol/L (P < 0.01). And, proportion of high and severe DNA damage increased with dose of HQ. HQ could also induce formation of micronucleus and abnormal nucleus in all groups treated by varied doses of HQ, with rates of micronucleus and abnormal nucleus of 2%, 3%, 10%, 9% and 15%, and 6%, 7%, 16%, 27% and 28%, respectively, in a significant dose-dependent manner. There was significant increase in rates of micronuclei and abnormal nuclei in cells treated with HQ at doses of 20, 40 and 80 micro mol/L (P < 0.05).

CONCLUSIONSExposure to HQ could cause DNA and nucleus damage inducing genotoxic effects on HLF.

Cell Nucleus ; drug effects ; Comet Assay ; DNA Damage ; drug effects ; Embryo, Mammalian ; Fibroblasts ; cytology ; Humans ; Hydroquinones ; toxicity ; Lung ; cytology ; Micronucleus Tests

OBJECTIVETo construct pEGFP-C1-T vector, an eukaryotic expression plasmid of hMTH1 gene antisense RNA.

METHODSThe conservative region of hMTH1 gene was amplified by RT-PCR after total RNA being extracted from human embryo lung fibroblast (HLF) and then cloned into pGEM-T vector. After the recombinant plasmid was certified by DNA sequencing, the conservative region of hMTH1 gene was inserted into pEGFP-C1 vector reversedly and pEGFP-C1-T vector was constructed. The efficiency of antisense inhibition was verified by Western blotting after cell transfection.

RESULTS423 bp fragment including conservative region of hMTH1 gene was obtained by RT-PCR. After cloned by pGEM-T vector and certified by DNA sequencing, pEGFP-C1-T vector was successfully constructed by means of recombinant DNA technology. Additionally pEGFP-C1-T vector could efficiently decrease hMTH1 protein level by 46%.

CONCLUSIONThe efficient expression vector of hMTH1 gene antisense RNA, pEGFP-C1-T has been constructed successfully.

DNA Repair Enzymes ; Genetic Vectors ; genetics ; Humans ; Phosphoric Monoester Hydrolases ; genetics ; Plasmids ; RNA, Antisense ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo detect the point mutations by chip-based capillary electrophoresis and to provide a rapid and sensitive technique detection for beta-thalassemia.

METHODSMultiplex primer-extension reaction was used to amplify the common loci of the samples for beta-thalassemia. The reaction products were detected by the chip-based capillary electrophoresis and the genotypes of the samples were discrened.

RESULTSA system was constructed to detect the point mutations of beta-thalassemia by chip-based capillary electrophoresis, and the technology was ralidated by the patients' samples and the results coincided with those of detection kit.

CONCLUSIONBeta-thalassemia can be detected by chip-based capillary electrophoresis rapidly with a small amount of samples. It would be a new detection method of the genetic disorders.

Electrophoresis, Capillary ; methods ; Female ; Genetic Diseases, Inborn ; diagnosis ; genetics ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; beta-Thalassemia ; diagnosis ; genetics

OBJECTIVETo construct DNA double-strand break (DSB) repair protein hKu70 deficient cell strain and to observe its biological characters for studying the functions of hKu70 gene and the effects of occupational harmfulness factors on DSB repair.

METHODSHuman lung fibroblasts (HLF) were transfected with the eukaryotic expression plasmids of hKu70 gene antisense RNA (pEGFP-C1-K) to construct hKu70 protein deficient cells (named as "HLFK"). The protein expression levels of hKu70 gene in HLFC and HLFK were detected by the Western blotting to estimate the effects of antisense inhibition. Morphology, growth character and growth status in soft agar of transfected HLFK were observed.

RESULTSpEGFP-C1-K vector was successfully expressed in HLF. The protein expression level of hKu70 gene in HLFK was decreased by 42% as compared with that in HLFC. No obvious changes of the biologic characters were observed in HLFK.

CONCLUSIONThe hKu70 protein deficient cell strain was successfully constructed. The hKu70 protein deficiency alone didn't induce obvious changes of the biological characters in HLFK.

Antigens, Nuclear ; analysis ; Cell Division ; DNA Damage ; DNA Helicases ; DNA Repair ; DNA-Binding Proteins ; analysis ; deficiency ; Humans ; Ku Autoantigen ; RNA, Antisense ; Transfection