OBJECTIVETo investigate the effect of compound Coptidis Rhizoma capsule (CCRC) on unbalanced expression of renal tissue TGF-β1/BMP-7 and Smad signaling pathway in rats with early diabetic nephropathy (DN), and discuss CCRC's effect on DN rats with early diabetic nephropathy and its possible mechanism.

METHODDN model rats were established by injecting streptozotocin (STZ). The rats were randomly divided into seven groups: the normal group, the model group, the enalapril treatment group, the xiaoke pill treatment group and three CRCC treatment groups. They were orally administered once a day for five weeks. The fasting blood glucose (FBG), blood urea nitrogen (BUN), serum creatinine (Scr), insulin (Ins), 24 h urinary protein (24 h Upro) and 24 h urinary microalbumin (24 h UmAlb) were tested. The pathological changes in renal tissues were examined by optical microscopy. Immuno- histochemical measures were used to detect the expressions of TGF-β1, BMP-7, Smad2/3, Smad1/5, and Smad7 protein, and RT-PCR was used to detect TGF-β1 mRNA and BMP-7 mRNA in renal tissues.

RESULTCompared with model group, BUN, Scr, Ins, 24 h Upro and 24 h UmAlb levels decreased at different degrees in CCRC treatment groups; the abnormal pathomorphology in renal tissue was improved; immunohistochemistry results showed that the expression of TGF-β1 and Smad2/3 were reduced, while the expression of BMP-7, Smad1/5 and Smad7 increased in CRCC treatment groups; the expression of TGF-β1 mRNA were reduced, but the expression of BMP-7 mRNA had no obvious change in CRCC treatment groups.

CONCLUSIONCRCC can improve the early renal function, delay the progression of chronic renal pathology and maintain the dynamic balance of TGF-β1/BMP-7 expression in renal tissues of DN rats. The mechanism may be related to down-regulation of renal TGF-β1 and up-regulation of BMP-7 through Smad signaling pathway.

Animals ; Bone Morphogenetic Protein 7 ; genetics ; metabolism ; Coptis ; chemistry ; Diabetic Nephropathies ; drug therapy ; genetics ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Kidney ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Rhizome ; chemistry ; Signal Transduction ; drug effects ; Smad Proteins ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo investigate the effect of a phosphorothioate antisense oligodeoxynucleotide "ASOND" combined with cis-Diamminedichloroplatinum (DDP), 5-fluorouracil (5-FU) and adriamycin (ADM) respectively on inhibiting the proliferation of HepG2 cells.

METHODSA phosphorothioate antisense oligodeoxynucleotide (5'-ACTCACTCAGG CCTCAGACT-3') targeted to human telomerase reverse transcriptase (hTERT) mRNA, which named cantide, was synthesized. ASODN was transfected into HepG2 by lipofectin. And cell growth activity was evaluated by MTT assay. SAS software and Jin Zhengjun Method were used to evaluate the interaction of ASODN and these chemotherapeutic drugs.

RESULTSCombination treatments with 0.1micromol/L ASODN reduced the IC50 of DDP, 5-FU and ADM from 1.07, 4.15 and 0.29microg/ml to 0.25, 1.52 and 0.12microg/ml respectively. The inhibitory ability of combination treatments on HepG2 cells was higher than that of these drugs alone (F=66.92, 25.96, 8.56, P<0.001). And synergism (Q>or=1.15) was observed at the lower concentration of DDP (

CONCLUSIONASODN may enhance therapeutic effectiveness of chemotherapeutic drugs in human hepatocellular carcinoma cells.

Antineoplastic Agents ; administration & dosage ; Cell Line, Tumor ; Cisplatin ; administration & dosage ; DNA-Binding Proteins ; Doxorubicin ; administration & dosage ; Drug Synergism ; Fluorouracil ; administration & dosage ; Humans ; Liver Neoplasms ; drug therapy ; Oligodeoxyribonucleotides, Antisense ; administration & dosage ; Telomerase ; antagonists & inhibitors ; genetics

AIM:To investigate the influence of pinellia heart-draining Decoction, Licorice heart-draining Decoction, Fresh ginger heart-draining Decoction and its dismantlements on the amount of gastric mucus in rats. METHODS:20 different combination of Banxiaxiexin Decoction and its analogus according to the unform design layout, the amount of gastric mucus in rats was determinated. Stepwise regression analysis was adopted to estimate the relationship between the pharmacological data and compatibility. RESULTS:Rhizoma Coptidis could reduce the amount of gastric mucus remarkbably, the interaction of Rhizoma Pinelliae、Fructus Ziziphi Jujubae and Radix Glycyrrhizae could increase the amount of gastric mucus, and interaction was confirmed among different medicinal ingredients in prescription. CONCLUSION:Uniform design provides a referable exeperimental design for studying analogus decoction compatibility law.

Objective To explore the role of Clara cell secretory protein(CCSP) in asthmatic children and compare the levels of CCSP in sera and induced sputum.Methods Thirty-four children with asthma who were in remission and 25 healthy controls were enrolled.Sera and hypertonic saline-induced sputum were obtained in asthmatic children,and sera alone were obtained in control subjects.The le-(vels) of CCSP were measured in sera and induced sputum by enzyme linked immunosorbent assay.Results Asthmatic children,compared with controls,had significantly lower concentration of CCSP in sera(P

Objective To evaluate the health-related quality of life (HRQOL) for prostate cancer patients receiving androgen deprivation therapy (ADT).Methods We studied 200 patients with prostate cancer who were treated with primary ADT in Peking University People's Hospital from June 2014 to June 2015.The patients'average age was 73.9 years.The mean PSA level was 21.57 ng/ml when they were diagnosed with prostate cancer.Of these 200 patients,79％ (158/200) were localized and seclected ADT therapy due to age,body condition,basic diseases or individual will.21％ (42/200) were locally advanced or metastasic,which accord with the indication of ADT therapy.The scales,including the MOS item short from health survey (SF-36),Functional Assessment of Cancer Therapy-General module (FACT-G),Functional Assessment of Cancer Therapy-Prostate instrument (FACT-P),Self-Rating Depression Scale (SDS) and Self-Rating Anxiety Scale (SAS) were used to evaluate their HRQOL.Results According to results of SF36,the physical health component score and mental health component score was 67.41 ± 16.39 and 64.81 ± 17.51,respectively.They indicated that the overall quality of life of these patients was at an acceptable level.And the average score of all domains exceeded 60 except general health domain,which the score was only 40.03 ± 21.89.When it comes to FACT-P,the sum score,emotional well-being score and prostate cancer component score were 8.15 ± 3.72,12.30 ± 4.04 and 77.41 ± 9.95,respectively,which were less than half of their respective top score.However the physical well-being score was 20.41 ±4.29,which was a relatively satisfactory value.In addition,61％ (121/200) patient selected not at all in item I am able to have and keep an erection.The score of SDS and SAS was 46.76 ± 8.29 and 43.25 ±9.69,respectively.And there were 23.0％ (46/200) and 21.5％ (43/200) patiens exhibited depression and anxiety,respectively.Conclusion In the present study,the prostate cancer patients receiving ADT showed acceptable HRQOL,but some patients sufferd from depression,anxiety and erectile dysfunction.

OBJECTIVETo investigate the effects of serum containing Gumibao (Chinese character: see text) on the proliferation and differentiation of osteoblast induced by dexamethasone.

METHODSOsteoblasts were extracted from skulls in newly born (within 24 hours) SD rats, and digested with collagenase. The first passage of cells were used for experiments. Cells were cultured in the medium containing different concentrations of dexamethasone (0, 10(-8), 10(-7), 10(-6), 10(-5) ,10(-4) mol/L). Alkaline phosphatase staining were carried out after 1 week and numbers of mineralized nodes with alizarin red staining were observed after 3 weeks. Accordingly, following the treatment of 10(-5) mol/L dexamethasone for 1 week, cells were cultured in the medium with serum containing Gumibao (Chinese character: see text). One week after Cumibao (Chinese character: see text) treatment, cells were stained with Alkaline phosphatase and collagen I and PCNA were examined by Western-blot. However, the observation of numbers of mineralized nodes with alizarin red stain required one more week.

RESULTSHigh concentration of dexamethasone could inhibit the expression of PCNA, collagen I, alkaline phosphatase and reduce the number of mineralized nodes of osteoblast, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

CONCLUSIONHigh concentration of dexamethasone could inhibit the proliferation and differentiation of osteoblastic cells, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; analysis ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Female ; Osteoblasts ; cytology ; drug effects ; physiology ; Proliferating Cell Nuclear Antigen ; analysis ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo establish a reliable model for drug screening and therapy by culturing rat femoral head and inducing cartilage degeneration quickly in vitro.

METHODSThe femoral heads from the same SD rats of two-month old were divided into control group and experimental group respectively. They were cultured with DMEM medium plus 10% fetal bovine serum or DMEM medium plus 10% fetal bovine serum plus 50 ng/ml IL-1β for three days. Femoral heads were fixed in 4% paraformaldehyde, decalcified, dehydrated, embedded in paraffin and cut into slices. Specimens were stained with Toluidine blue and Safranine O-Fast Green FCF. The protein expression levels of type II collagen, MMP13, Sox9 and ADAMTS5 were analyzed by immunofluorescence.

RESULTSBoth the Toluidine blue and Safranine O staining were pale in the margin of femoral heads which were stimulated with IL-1β for three days compared to that in control group. The Fast Green FCF staining was positive at the edge of the femoral head in experimental group, which indicated that cartilage became degenerated. The expression levels of both type H collagen and Sox9 were decreased significantly while the expression levels of MMP13 and ADAMTS5 were increased in experimental group.

CONCLUSIONThe model of cartilage degeneration is established by culturing and inducing the degeneration of the femoral heads quickly in vitro.

Animals ; Cartilage Diseases ; genetics ; metabolism ; Collagen Type II ; genetics ; metabolism ; Disease Models, Animal ; Femur Head ; metabolism ; Humans ; In Vitro Techniques ; Interleukin-1beta ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 13 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; SOX9 Transcription Factor ; genetics ; metabolism

OBJECTIVETo investigate an safe and effective new technology (treatment) to repair maxillofacial region penetrating defect.

METHODSThe lower trapezius musculocutaneous flap is parallel just like as two leaves which is connected to each other, and was folded to provide the liner of oral cavity and external cover.

RESULTSTotally twelve folding lower trapezius musculocutaneous pedicle flap survived. Postoperative follow-up for 1 approximately 3 years, the patients restored the function as well as the shape of maxillofacial region.

CONCLUSIONSThe lower trapezius musculocutaneous pedicle flap is a suitable material for maxillofacial region reconstruction, further more, the successful rate is perfect.

Female ; Humans ; Male ; Mouth Neoplasms ; surgery ; Muscle, Skeletal ; transplantation ; Oral Surgical Procedures ; methods ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; methods ; Surgical Flaps ; Treatment Outcome

OBJECTIVETo compared the activity and yield rate of osteoblast obtained by different collagenase digestion methods, to find a better way to extract osteoblast for the experimental researches of osteoporosis.

METHODSTen 24-hour-old SD rats were were euthanized. The cranium of rats were removed and cuted into blocks of 1 mm x 1 mm size. After digested by trypsin for 15 min, all the cranium were divided into two equal parts, and randomly divided into two groups which would be digested by type I collagenase and type II collagenase separately for two times. The rat cells of the two groups were cultured in thermostat incubator with 5% CO2 under the condition of 37 degrees C. The primary culture osteoblasts were counted by using a haemacytometer after digestion and 72 hours later. The second generation osteoblasts cultured 48 h were dyed by NBT/BCIP staining solution, and were detected by quantitative measurement with PNPP.

RESULTSThe cells had irregular shapes. The results of cell counting showed that the cell number of type I group was larger than type 11 group. Alkaline phosphatase dyeing were positive. Detecting of alkaline phosphatase using the method of PNPP showed that the absorbance value in type I group were higher than type II group (P<0.05).

CONCLUSIONTwo types of collagenase are both suitable for the in vitro culture of rat osteoblasts. The activity and yield rate of osteoblasts in type I group are higher which could provide more stable seed cells for the treatment of osteoporosis.

Animals ; Cell Count ; Cell Culture Techniques ; methods ; Collagenases ; metabolism ; Female ; Male ; Osteoblasts ; cytology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo detect the effect of connective tissue growth factor (CTGF) on podocalyxin expression in mouse podocytes exposed to high glucose in vitro and explore the possible pathway involved.

METHODSThe expression vector carrying a small interfering RNA (siRNA) targeting CTGF was transfected into mouse podocytes cultured in the presence of 1 g/L glucose (normal control), 4.5 g/L glucose (high glucose group), 1 g/L glucose + 3.5 g/L mannitol (iso-osmolar control group). The changes in the protein expression levels of podocalyxin, CTGF and ERK1/2 in the cells in response to the treatments were investigated using Western blotting.

RESULTSHigh glucose exposure for 24 and 48 h resulted in significantly decreased expression of podocalyxin and increased CTGF in the podocytes (P<0.05). Phosphorylation of ERK1/2 occurred as early as 30 min after the exposure, and the activation was maintained till 24 h. Transfection of the cells with siRNA targeting CTGF significantly inhibited these changes.

CONCLUSIONCTGF is an important mediator of high glucose-induced podocyte damage and decreases the protein level of podocalyxin by the ERK1/2 pathway. CTGF-specific siRNA can alleviate high glucose-induced podocyte injury, suggesting its potential value in treatment of diabetic nephropathy.

Animals ; Cells, Cultured ; Connective Tissue Growth Factor ; metabolism ; Diabetic Nephropathies ; Glucose ; adverse effects ; MAP Kinase Signaling System ; drug effects ; Mice ; Podocytes ; cytology ; drug effects ; metabolism ; RNA, Small Interfering ; genetics ; Sialoglycoproteins ; metabolism