This study retrospectively reviewed the experimental clinical programs at The First Affiliated Hospital of Nanjing Medical University. Experience was summarized, and methods for improving clinical pharmaceutical research were discussed including recruiting talent people, managing resources and trail results, and establishing international cooperations.

OBJECTIVETo investigate the antagonizing effect of Hirsutella sinensis (HS) on renal tubular epithelial-myofibroblast transdifferentiation (TEMT) and its possible pathogenic mechanism in rats with chronic aristolochic acid nephropathy (CAAN).

METHODSEighteen male Sprague-Dawley rats were equally divided into 3 groups, the model (M) group, the intervention (I) group and the control (C) group. The 24 h urinary protein (UP) in rats was measured before intervention and at the end of the 1st, 4th, 8th, and 12th week, and creatinine clearance rate (CCr) was measured before intervention and at the end of the 12th week respectively. All rats were sacrificed at the end of the 12th week, their kidney was taken for examining the degree of fibrosis in renal interstitial with Masson's stain and determining mRNA and protein expressions of transforming growth factor-beta1 (TGF-beta1), Snail, alpha-smooth muscle actin (alpha-SMA) and cytokeratin in renal tissue by Real-time RT-PCR and immunohistochemistry staining, respectively.

RESULTSCompared with the C group, CCr was significantly lower, while 24 h UP was higher; the relative area of interstitial fibrosis was significantly larger in the M group; besides, the mRNA and protein expressions of TGF-beta1, Snail and alpha-SMA were significantly up-regulated (P < 0.01 or P < 0.05), and those of cytokeratin were significantly down-regulated (P < 0.01) in renal tissue of the M group. While in the I group, all the above-mentioned abnormalities were restored to some extent (P < 0.05) and showed significant difference (all P < 0.05) as compared with those in the M group.

CONCLUSIONHS can downregulate TGF-beta1 and Snail expressions in renal tissue, antagonize TEMT and renal interstitial fibrosis, and improve renal function in CAAN rats.

Actins ; genetics ; metabolism ; Animals ; Aristolochic Acids ; toxicity ; Cell Transdifferentiation ; Chronic Disease ; Cordyceps ; chemistry ; Drugs, Chinese Herbal ; therapeutic use ; Fibroblasts ; drug effects ; Kidney Diseases ; chemically induced ; metabolism ; Kidney Tubules ; pathology ; Male ; Phytotherapy ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Snail Family Transcription Factors ; Transcription Factors ; genetics ; metabolism ; Transforming Growth Factor alpha ; genetics ; metabolism

Objective To investigate the prevalence for endemic fluorosis of drinking water type and to discuss the relationship between endemic fluorosis and urinary fluorine in Linyi county, Shanxi province. Methods In 2006, three counties were selected as heavy, medium and control areas according to the distributing feature of the disease. The dental fluorosis in each spots was examined by Dean method. The levels of urinary fluorine were determined by fluorine selective ion electrode. The skeletal fluorosis of adults were examined by X-ray. Results There was evident differences of dental fluorosis and skeletal fluoresis among the heavy and the medium endemic fluorosis and control areas(X~2 = 410.945, P < 0.01 ), the prevalence of dental fluoresis in the medium area and the heavy area were 92.34% (253/274), 90.09% ( 291/323), significantly higher than in the control area[23.27% (64/275), X~2 = 274.927,268.287, all P < 0.01]. The heavy area had the highest rate of the skeletal fluorosis rate [59.75% (141/236) ], the medium area had the middle-level of the skeletal fluorosis rate[24.76%(52/210), X~2 = 183.578, P< 0.01]. Urine fluorine contents in both beavy[ (4.69 ± 0.17)mg/L] and medium areal (4.86 ± 0.13)mg/L] were higher than that in the control areas[ (1.75 ± 0.04)mg/L, H = 411.197, P< 0.01], and there was linear relevance between the different degree of skeletal fluorosis and urine fluorine contents (r = 0.508, P < 0.01). Conclusions The local fluoresis condition of Linyi county in Shanxi province was serious. The degree of skeletal fluorosis is associated with the fluoride content in urine.

Angiography, Digital Subtraction ; Cranial Nerve Neoplasms ; diagnosis ; diagnostic imaging ; surgery ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Paraganglioma ; diagnosis ; diagnostic imaging ; surgery ; Vagus Nerve Diseases ; diagnosis ; diagnostic imaging ; surgery

Two hundred thirty specimens of wild birds were collected from some areas in Heilongjiang Province during the period of 2003~2004, including two batches of specimens collected randomly from a same flock of mallards in Zhalong Natural Reserve in August and December, 2004, respectively. Primary virus isolation and identification for avian influenza virus (AIV) and Newcastle disease virus (NDV) were performed. The results showed that only two specimens of young mallards collected from Zhalong Natural Reserve in August, 2004 were positive to AIV (isolation rate 0.9%), and one strain (D57) of these two virus isolates was identified to be H9 subtype by hemagglutination inhibition test. Meanwhile, the two batches of blood serum samples of mallards from Zhalong were also examined for antibodies against AIV and NDV. Among 38 blood serum samples collected in August, antibodies against the hemagglutinin of H1, H3, H5, H6 and H9 subtypes of AIV were found in 1, 0, 2, 0 and 8 samples, respectively; and 11 samples were found with antibody against NDV. Whereas the NDV isolation in both two batches of specimens of mallard was negative, all of the 32 blood serum samples collected in December were negative for antibodies against AIV and NDV.
Animals ; Animals, Wild/*virology ; Antibodies, Viral/isolation&purification ; Birds/virology ; China/epidemiology ; Hemagglutination Tests ; Influenza A virus/*isolation&purification ; Influenza in Birds/epidemiology/immunology/*virology ; Newcastle Disease/epidemiology/immunology/*virology ; Newcastle disease virus/*isolation&purification ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo observe the expressions of caspase-8 and caspase-9 mRNA, and explore the changes of apoptosis of bone marrow hematopoietic cells in patients with chronic mountain sickness (CMS).

METHODSOf 18 CMS patients and 16 controls were enrolled in this study. The apoptotic index (AI) of bone marrow mononuclear cells (BMMNC) was measured by TUNEL technique, the levels of caspase-8 and caspase-9 mRNA in BMMNC of CMS patients and controls were determined by RT-PCR. Results (1)The AI of BMMNC in patients with CMS (8.51 ± 3.35)% was lower than that in controls (16.00 ± 4.28)% (P < 0.01); (2) The values of caspase-8 and caspase-9 mRNA were (0.28 ± 0.07) and (0.23 ± 0.08) respectively, in CMS patients, which were significantly lower than those of (0.45 ± 0.09) and (0.41 ± 0.09) respectively, in the controls (both P < 0.01); (3) Hemoglobin (Hb) value was negatively correlated with levels of caspase-8 and caspase-9 mRNA (r values were -0.52 and -0.61 respectively, both P < 0.05) in CMS patients. There was a negative correlation between AI and Hb (r value was -0.89, P < 0.01) in CMS patients. However, the significant relationship was not found between AI and level of caspase-8 or caspase-9 mRNA (P > 0.05).

CONCLUSIONSThe results showed a decrease apoptosis of BMMNCs and reduced levels of caspase-8 and caspase-9 mRNA in CMS patients, the latter might be involved in the change of BMMNCs apoptosis.

Adult ; Altitude Sickness ; metabolism ; pathology ; Apoptosis ; Bone Marrow Cells ; metabolism ; Case-Control Studies ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Humans ; Male ; Middle Aged

OBJECTIVETo construct of tissue engineering skin including active composite dermal matrix.

METHODSThe human fibroblasts and bovine collagen with type I were inoculated on the surface of porcine acellular dermal matrix (PADM) for construction of active dermal substitute, then epidermal cells were inoculated on the dermal matrix for gas-liquid interface culture. The tissue-engineering skin was observed by histological examinations.

RESULTSThe structure of fibroblasts in collagen was intact, which was used to construct composite dermal matrix with PADM. The epithelial structure of tissue-engineering skin was similar to that of normal skin with good cell differentiation. Some phenomena were showed in epidermis: basic layer, stratum spinosum, granular layer and stratum corneum, desmosomes.

CONCLUSIONFibroblasts-Collagen-PADM can be an optimal dermal matrix for construction of tissue-engineering skin.

Animals ; Cattle ; Cell Culture Techniques ; Collagen Type I ; Dermis ; transplantation ; Epidermis ; cytology ; Extracellular Matrix ; transplantation ; Fibroblasts ; cytology ; Humans ; Skin ; cytology ; Skin, Artificial ; Swine ; Tissue Engineering

OBJECTIVETo investigate changes in apoptosis-related ligands in serum in rats with severe scald and the effect of intensive insulin therapy on the changes.

METHODSOne hundred and fifty Wistar rats were randomly divided into 3 groups: sham burn (SB), scald (S) and treatment (T) groups. Rats in S and T groups were inflicted with 40% TBSA full-thickness burn, followed by intraperitoneal injection with 40 mL/kg of isotonic saline for resuscitation. Rats in T group were subcutaneously injected insulin in a dose of 0.25 U/100 g 24 hours after burn injury, and every 12 hours for 5 days (0.25, 0.50, 0.75, 1.00, 1.25 U/100 g each day, respectively) to control the level of blood glucose between 3 and 6 mmol/L. Rats in SB group were sham scalded at 37 degrees C without resuscitation. Blood was drawn from abdominal aorta on 1, 4, 7, 10, 14 post burn day (PBD) for determination of serum levels of TNF-alpha, soluble Fas ligand (sFasL) and soluble Fas receptor (sFas) by enzyme-linked immunosorbent assay (ELISA), and insulin by radioimmunity assay (RIA).

RESULTSThe serum level of TNF-alpha in S group peaked on 1 PBD (30.9 +/- 8.7) ng/L, which showed statistically significant difference when compared with that of SB and T groups (12.7 +/- 2.8) ng/L, (16.8 +/- 4.7) ng/L, respectively, P < 0.01), then lowered gradually to become similar to that of SB group on 7 PBD. The level of TNF-alpha in T group increased gradually, but was obviously lower than that of S group on 1, 4, 7 PBD (P < 0.01). The level of sFasL in S (on 7-14 PBD) and T (4-10 PBD) groups was significantly higher than that in SB group (P < 0.05), then lowered to normal level. The levels of sFas on 4-10 PBD in T group were obviously higher than that in S and SB group (P < 0.05). Ratio of sFasL to sFas in serum of S group was higher than that in SB group on 7, 10 PBD, which was higher than that in T group on 7 PBD (P < 0.05). There was significant decrease in serum level of insulin in S group compared with that of SB group on 4-10 PBD (P < 0.05). The level of insulin in T group increased on 1 PBD, peaked on 4 PBD (327 +/- 15 microU/mL), which was significantly higher than that in SB and S groups (42 +/- 15, 28 +/- 10 microU/mL, respectively, P < 0.01), then decreased gradually to normal level.

CONCLUSIONSInsulin may inhibit apoptosis after burn by down-regulating secretion of apoptotic ligands.

Animals ; Apoptosis ; Blood Glucose ; analysis ; Burns ; blood ; drug therapy ; Fas Ligand Protein ; blood ; Insulin ; therapeutic use ; Male ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; blood ; fas Receptor ; blood

Objective To investigate the seroprevalence of tick-borne diseases in humans and domestic animals from rural areas of Zhejiang province. Methods Anji county, Jindong district and Tiantai county were selected for samples collection according to their geographic locations and historical prevalence of tick-borne diseases. Blood samples of humans and domestic animals were collected in the three sites. An indirect immuno-fluorescent antibody test was used to determine the presence of IgG antibodies of Rickettsiae heilongjiangii, Orientia tsutsugamushi, R. typhi, Anaplasma phagocytos, Ehrlichia chaffeensis, Bartonella, R. hainan and Coxiella burnetii in these samples.Results Six hundred and eighty-three blood samples including 579 from humans and 104 from domestic animals(53 from cattles and 51 from sheep)were collected from the three sites. Antibody positive rates of Orientia tsutsugamushi, R. typhi, Ehrlichia chaffeensis and Coxiella burnetii were significantly different between these sites. IgG from all the 8 pathogens were detected in samples from humans. It was found that the sero-prevalence rates of R. typhi, Bartonella and C. burnetii(20.7%,10.9%, 5.5%)of adults were higher than those of other Rickettsiae under investigation. The seroprevalence of R. typhi increased along with age. IgG from the 7 pathogens were detected in samples from domestic animals except for Anaplasma phagocytos. The sero-prevalence rates of R. typhi, Bartonella and R. hainan(69.2%, 51.0%, 22.1%)of adults were higher than those of other Rickettsiae investigated. Conclusion Tick-borne diseases did spread widely in humans and domestic animals from different rural areas of Zhejiang province. The sero-prevalence rates of R. typhi,B. henselae, R. hainan and C. burnetii were higher than that from other pathogens.

OBJECTIVETo prepare a porcine acellular dermal matrix (PADM), and to optimize the interpore distance between PADM and co-grafted split-thickness autologous skin.

METHODSPorcine skin was treated with trypsin/Triton X-100 to prepare an acellular dermal matrix. Micropores were produced on the PADM with a laser punch. The distance between micropores varied as 0.8 mm, 1.0 mm, 1.2 mm and 1.5 mm. Full-thickness defect wounds were created on the back of 144 SD rats. The rats were randomly divided into 6 groups as follows, with 24 rats in each group. Micropore groups I -IV: the wounds were grafted with PADM with micropores in four different intervals respectively, and covered with split-thickness autologous skin graft. Mesh group: the wounds were grafted with meshed PADM and split-thickness autograft.

CONTROL GROUPwith simple split-thickness autografting. The gross observation of wound healing and histological observation were performed at 2, 4, 6 weeks after surgery. The wound healing rate and contraction rate were calculated.

RESULTSTwo and four weeks after surgery, the wound healing rate in micropore groups I and II was lower than that in control group (P < 0.05), but no obvious difference was between micropore groups I , II and mesh group (P > 0.05) until 6 weeks after grafting( P <0.05). The wound contraction rate in micropore groups I and II ([(16.0 +/- 2.6)%, (15.1 +/- 2.4)%] was remarkably lower than that in control group 4 and 6 weeks after grafting (P < 0.05), and it was significantly lower than that in mesh group [(19.3 +/- 2.4)%] 6 weeks after surgery (P <0.05). Histological examination showed good epithelization, regularly arranged collagenous fibers, and integral structure of basement membrane.

CONCLUSIONLaser micropore PADM (0.8 mm or 1.0 mm in distance) grafting in combination with split-thickness autografting can improve the quality of wound healing. PADM with laser micropores in 1.0 mm distance is the best choice among them.

Animals ; Dermis ; transplantation ; Lasers ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; methods ; Skin, Artificial ; Swine ; Transplantation, Heterologous