Mycoepoxydiene is a novel antitumor agent extracted from marine lignicolous fungi HLY-2, which is Diaporthe phaseolorum by molecule identification. The medium optimization for mycoepoxydiene by orthogonal design and the comparison of submerged fermentation and solid state fermentation were studied. The rusult is that the maximal yield of the compound is 543mg/L, which is 43 times compared to the customary half-seawater PD medium and 15 times to the best submerged condition. This optimum culture medium included potato 250g/L, seawater 300mL/L, glucose 30g/L, lactose 50g/L, KH_ 2 PO_ 4 0.65mmol/L and (NH_ 4 )_ 2 SO_ 4 1g/L in the solid state condition. Differentiation analysis between submerged and solid state fermentation, and antitumor activity of these ferment products were also studied. The antitumor activity of products of the optimum medium approached the pure compound.

One hundred and seventy-two strains of endophytic fungi were isolated from Taxus mairei,Cephalotaxus fortunei and Torreya grandis cv.merrillia.The result of the antifungal assay shows that ninety strains of the fungi have antagonism against one or more botanical pathogenic fungi,such as Neurospora sp.,Trichoderma sp.,Fusarium sp.etc.The percentage of antifungal strains to tested strains are as follows:40% Cephalotaxus fortunei,54.2% Taxus mairei,57.1% Torreya grandis cv.merrillia.Thirty-five strains have high antifungal activities,and their inhibition zone diameter is at least 15mm.The active endophytic fungi were identified as 18 genera,most of which belong to Paecilomyces and Fusarium etc.

BACKGROUND: It has been proved that the application of basic fibroblast growth factor (bFGF) combined with Brucea Javanica oil emulsion can accelerate wound healing and inhibit scar formation.OBJECTIVE: To observe the effects of bFGF plus Brucea Javanica oil emulsion cream on accelerating the skin wound healing of rabbits.DESIGN: A randomized controlled observation.SETTING: Center of Biotechnological Research and Development, Jinan University; College of Pharmacy, Jinan University.MATERIALS:The experiment was carried out in the College of Pharmacy, Jinan University from June to September in 2004. Eight Beijing big-ear white rabbits (4 males and 4 females) of 2.0-2.5 kg were provided by the experimental animal center of Southern Medical University (certification number: SoKx-2002-010). bFGF sterile freeze dried powder agent,provided by Guangzhou Changsheng Gene Pharmaceutical Co.,Ltd (batch number: 20040219; specific activity was 6 000 U/bottle), was prepared to solution with water for injection before application. Brucea Javanica oil emulsion (manufactured by Zhejiang 999 Bang'erkang Pharmaceutical Co.,Ltd.) was provided by Professor Yao from staff Room of Pharmacy, Shenyang Pharmaceutical University.METHODS: The rabbits were anesthetized and disinfected, 5 round wounds with diameter of 1.8 cm and area of 2.54 cm2were induced from front to back by bilateral incision at 1.5 cm from middle spine of rabbit. The 5 wounds of each rabbit were randomly divided into bFGF-treated group (90 U/cm2), bFGF+Brucea Javanica oil emulsion group [the wound was smeared with Brucea Javanica oil emulsion (30 mg/cm2) 30 minutes after bFGF (90 U/cm2)], Brucea Javanica oil emulsion treated group [the wound was smeared with Brucea Javanica oil emulsion (30 mg/cm2)], blank emulsion group (30 mg/cm2) and blank control group (the wound was smeared with saline). The medication was give immediately after injury, and changed once a day for 16 days. At 4, 8, 12 and 16 days after injury, the wound areas were recorded with the method of hyaline membrane tracing (the wound was covered with clean saran wrap, the size of wound was traced,and then sheared to be weighed, and converted to calculate the area), and the volume of wounded cavity was measured by infusing water. At 8 and 16 days, the wound tissue was removed, stained after routine tissue sections, and the conditions of growth of granulation tissue and reepithelization on the wound surface were observed.MAIN OUTCOME MEASURE: The wound area, volume of wounded cavity, and the conditions of growth of granulation tissue and reepithelization on the wound surface were obviously at different time points after injury in each group.RESULTS:All the 8 rabbits were involved in the analysis of results. ① Wound areas at different time points in each group: The wound areas in the bFGF+Brucea Javanica oil emulsion group at 4, 8 and 12 days after medication were smaller than those in the blank control group at corresponding time points [(2.05±0.35), (1.59±0.25), (0.55±0.25) cm2;(2.53±0.30), (2.41±0.19), (1.09±0.34) cm2, P＜0.05-0.01]. The wound areas in the bFGF group at 8 and 12 days after medication were (1.71±0.31) and (0.51±0.10) cm2, which were significantly smaller than those in the blank control group at corresponding time points (P＜0.05-0.01). ② Volume of the wounded cavity in each group: The wound volume in the bFGF group at 4 days after injury was markedly smaller than that in the blank control group at corresponding time point [(0.49±0.12), (0.59±0.1) mL, P＜0.05]. The wound volumes in the bFGF+Brucea Javanica oil emulsion group at 4 and 8days after injury were significantly smaller than those in the blank control group at corresponding time points [(0.47±0.12), (0.30±0.08) mL; (0.59±0.1), (0.41±0.07) mL, P＜0.05, 0.01]. ③ Growth of granulation tissue and reepithelization on the wound surface in each group: At 8 days after injury, the inflammatory reaction was milder and fibroblasts proliferated significantly in the bFGF+Brucea Javanica oil emulsion group, and the numbers of capillary plumules and fibroblasts were significantly more than those in the blank control group. The conditions in the blank control group and blank emulsion group were generally the same that there were severe inflammatory reactions, obvious increase of granulation tissue, fewer new capillaries, and unobvious proliferation of epidermic cells. At 16 days after injury, the contraction and reepithelization on the wound surface were obvious, and the new epithelia went towards the wound center rapidly in the bFGF+Brucea Javanica oil emulsion group.CONCLUSION : The application of Brucea Javanica oil emulsion plus bFGF can obviously accelerate the repair of incised wound on the back of rabbits.

Objective To evaluate the recognition and uptake of transglutaminase 3 (TG3) by dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN) receptors on the membrane surface of DC-SIGN-transfected human embryonic kidney (HEK) 293T cells and monocytederived dendritic cells (MDDCs).Methods The eukaryotic expression vector pGCMV-enhanced green fluorescent protein (EGFP) containing DC-SIGN gene fragments was transfected into HEK293T cells to prepare DC-SIGN-EGFP-HEK293T cells by using liposome transfection method.CD14+ monocytes were isolated from peripheral blood samples by magnetic bead-based negative selection,and then were induced by granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) to prepare MDDCs.Laser confocal microscopy and flow cytometry were performed to evaluate the recognition and uptake of TG3 protein by DC-SIGN receptors on the surface of HEK293T cells and MDDCs.MDDCs treated without Alexa Fluor 647 dye-tagged TG3 served as blank control group,and those treated with Alexa Fluor 647 dye alone served as negative control group.Results After co-culture with TG3 for 3 hours,laser confocal microscopy and flow cytometry both showed that TG3 could be recognized by and uptaken through DC-SIGN receptors into HEK293T cells and MDDCs.Flow cytometry also revealed that the binding of TG3 to MDDCs could be partially blocked by DC-SIGN blocking antibodies.Neither the negative control group nor the blank control group showed the recognition and binding of TG3 to HEK293T cells and MDDCs.Conclusion TG3 can serve as a kind of autoantigen to be recognized and bound by DC-SIGN receptors,followed by uptake by dendritic cells.

BackgroundDiabetic retinopathy (DR) is the result of the cytokine network disorders,the imbalance of angiogenic factor and vascular inhibitory factor is the start factor.ObjectiveTo analyze the levels of CXC chemotatic factors of type 2 diabetes mellitus patients,evaluate the clinical application value of them in different clinical types of DR using receiver operating characteristic (ROC)analysis and to approach the new way of individualized treatment.Methods This was a prospective research.The gold standard was ophthalmolscope and fundus fluorescein angiography.The levels of CXC chemotatic factors and multiplicaiton factors were measured in 96 cases with type 2 diabetes mellitus (66 cases with retinopathy and 30 cases without retinopathy as control).The assessment tasks were performed for these index and courses of DR with ROC curve.Results The expression of age,course of disease has significant difference in different courses of DR ( F =8.507,P =0.001 ; F =28.143,P =0.000).Compared with the control group,the expression of growth-related oncogene-α ( GROα ) ( t =- 2.172,P =0.035,AUC =0.625 ),whole blood viscosity 200 ( t =- 3.724,P =0.001,AUC =0.904 ) and neutrophilic leukocyte (t=-2.562,P =0.013,AUC =0.577 ) has significant difference in the group of mild NPDR.Compared with the control group,the expression of interferon-γ-inducible protein 10 ( IP-10 ) ( t =-3.591,P =0.001,AUC =0.592 ),platelet derivation growth factor-BB ( PDGF-BB ) ( t =- 3.233,P =0.003,AUC =0.735 ),vascular endothelial growth factor(VEGF) ( t =- 3.617,P =0.001,AUC =0.776 ),C peptide ( t =- 3.366,P =0.002,AUC =0.962 ),leukocyte ( t=-3.201,P =0.003,AUC =0.852) and neutrophilic leukocyte(t =-4.201,P=0.000,AUC =0.852) has significant difference in the group of moderate and severe NPDR.ConclusionsCXC chemotatic factors may act as reactivator in the pathogenesis of DR,GROα and IP-10 may be useful for clinical monitoring of the severity of DR,and evaluating the imbalance state of chemotatic factors maybe a new approach to clinical monitoring and prognosis of DR.

Objective To detect the serum level of transglutaminase 2 (TG2)-specific IgE (slgE) in patients with atopic dermatitis (AD),and to analyze its correlation with the disease condition.Methods A total of 77 patients with AD were enrolled into this study,including 44 patients aged ≥ 12 years and 33 patients aged ＜ 12 years.Of the 77 patients,20 were diagnosed with intrinsic AD,which was characterized by the absence of sIgE and total serum IgE values ＜ 150 kU/L,and 49 with extrinsic AD characterized by positive (++) or even strongly positive slgE for more than 1 kind of exogenous allergens,or total serum lgE values ≥ 150 kU/L.[mmunocapture-biotinylated detector enzyme immunoassay was performed to detect the serum level of TG2-sIgE in 77 patients with AD,40 adult patients with psoriasis vulgaris (PV) and 30 healthy adult controls.Clinical data on the AD patients were recorded,including age,disease duration,SCORAD score,blood eosinophil count,levels of total IgE and TG2-sIgE.Results The serum levels of TG2-sIgE in AD patients aged ≥ 12 years,AD patients aged ＜ 12 years,PV patients and healthy controls were 1.02 ± 0.2,1.04 ± 0.044,0.93 ± 0.25 and 0.71 ± 0.13,respectively.Additionally,the serum level of TG2-sIgE significantly differed among AD patients aged ≥ 12 years,PV patients and healthy controls (x2 =37.407,P ＜ 0.001),and was significantly higher in both AD patients aged ≥ 12 years and PV patients than in the healthy controls (t =7.38,4.83,respectively,both P ＜ 0.001).Moreover,the intrinsic AD group showed significantly higher TG2-sIgE levels compared with the extrinsic AD group (1.16 ± 0.03 vs.1.02 ± 0.20,t =2.27,P =0.02).The TG2-sIgE level was uncorrelated with the patients' age,disease duration,SCORAD score,blood eosinophil count or serum total IgE levels in AD patients (r =0.03,0.14,-0.04,-0.08,0.06,respectively,all P ＞ 0.05).Conclusion The serum level of TG2-sIgE obviously increases in AD patients,so TG2 may be one kind of autoantigen in AD patients,but there is no significant correlation between the TG2-sIgE level and AD severity.

Objective To evaluate the clinical application and significance of lengthen-stem bipolar-femur prosthetic replacement for the treatment of old-age patients with intertrochanteric fracture osteoporosis.Methods 28 cases of patients aging from 75 to 99 years old of intertrochanteric fracture osteoporosis treated with lengthen- stem cemented bipolar prosthesis were studicd from March 2000 to December 2006.After taking the blank margin, the bones of different sizes were replaced and the steel wire was fixed.After determining the depth of the front an- gle,the artificial bone was placed.Results After 28 examples attaining the following-up examination for 7 months to 3 years,with an average of 1.5 years,its function according to Harris standard was evaluated 3 months after the operation,8 examples were excellent,13 examples good,5 examples pass,2 examples inferior.The excellent or good rate reached 75% ,with no abnormal cases,no joint dislocation during the followed-up period.1 example had the phantom phenomenon 1 year after the operation.2 examples among the inferior had got more serious internal medicine disease which affected the restoring function.1 example died of the internal medicine disease 1 year after the operation.Conclusion By using the lengthen-stem bipolar-femur prosthetic replacement for the treatment of old patients with inrertrochanteric fracture osteoporosis,the patients will restore quickly after the operation and can carry a heavy load at an early time.The illness complication and the mortality rate will be redaced.But its related disease must be strictly dealt with and the surgery operating skill must be grasped.

OBJECTIVETo study the antiviral constituents in the stems and leaves of Pithecellibium clypearia.

METHODThe constituents of P. clypearia were systematically separated with various chromatographic techniques in combination with antiviral activity monitoring. Their structures were elucidated by physical and chemical properties and spectral data.

RESULTSix compounds were isolated from P. clypearia and were identified as: tricetiflavan (5, 7, 3', 4', 5'-pentahydroxylflavan) (1), myricitrin (myricetin-3-O-alpha-L-rhamnopyranoside) (2), quercitrin (quercetin-3-O-alpha-L-rhamnopyranoside) (3), quereetin (4), methyl gallate (5) and gallic acid (6).

CONCLUSIONCompound 1 approximately 5 were obtained from this plant for the first time. Compound 4 was found to show an obvious anti-respiratory syncytial virus (RSV) activity.

Antiviral Agents ; chemistry ; isolation & purification ; pharmacology ; Fabaceae ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; pharmacology ; Gallic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; Inhibitory Concentration 50 ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; chemistry ; isolation & purification ; pharmacology ; Respiratory Syncytial Viruses ; drug effects

AIMTo study the expression pattern of the retinoic acid metabolizing enzymes RALDH2 and CYP26b1 during mouse postnatal testis development at both mRNA and protein levels.

METHODSReal-time polymerase chain reaction and Western blot analysis were performed to determine the relative quantity of RALDH2 and CYP26b1 at both mRNA and protein levels at postnatal day 1, 5, 10, 20, and in adult mice (70 days testes). Testicular localization of RALDH2 and CYP26b1 during mouse postnatal development was examined using immunohistochemistry assay.

RESULTSAldh1a2 transcripts and its protein RALDH2 began to increase at postnatal day 10, and remained at a high level through postnatal day 20 to adulthood. Cyp26b1 transcripts and CYP26b1 protein did not change significantly during mouse postnatal testis development. RALDH2 was undetectable in the postnatal 1, 5 and 10 day testes using immunohistochemistry assay. At postnatal day 20 it was detected in pachytene spermatocytes. Robust expression of RALDH2 was restricted in round spermatids in the adult mouse testis. In the developing and adult testis, CYP26b1 protein was confined to the peritubular myoepithelial cells.

CONCLUSIONOur results indicate that following birth, the level of retinoic acid in the seminiferous tubules might begin to increase at postnatal day 10, and maintain a high level through postnatal day 20 to adulthood.

Aldehyde Oxidoreductases ; genetics ; metabolism ; Animals ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Gene Expression Regulation, Developmental ; Male ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; metabolism ; Rabbits ; Retinoic Acid 4-Hydroxylase ; Seminiferous Epithelium ; cytology ; metabolism ; Sensitivity and Specificity ; Spermatids ; cytology ; metabolism ; Testis ; cytology ; growth & development ; metabolism

OBJECTIVETo observe the suppressive effect on the expression of pro-inflammatory cytokines in liver from brain dead (BD) rats through inhibition of p38 mitogen-activated protein kinase (MAPK) signaling pathway by SB203580.

METHODSA total of 30 male Wistar rats weighing from 180 to 200 g were randomly divided into 3 experimental groups: (1) BD group (n = 10): brain death was induced in rats; (2) BD+SB203580 group (n = 10): brain death was successfully induced and SB203580 (10 mg/kg) was given through dorsal vein of penis. After brain death artificial ventilation was maintained for 6 hours and only those with mean arterial blood pressure more than 80 mm Hg (1 mm Hg = 0.133 kPa) were accepted as BD donors. (3) Control group (n = 10): living healthy rats. The expressions of TNFalpha and IL-1beta mRNA in liver tissues were analyzed by RT-PCR and the protein expressions of TNFalpha, IL-1beta and phosphorylated p38MAPK were detected by Western blot.

RESULTSThe phosphorylated p38MAPK detected in the liver in BD group was significantly increased compared with the control group (q = 172.53, P < 0.01), and the expressions of TNFalpha and IL-1beta mRNA and proteins in liver were also significantly increased in BD group compared with the control group (q = 123.99, 135.35, 243.09 and 192.23, respectively, P < 0.01). The phosphorylated p38MAPK was decreased in BD+SB203580 group and significantly decreased compared with the BD group (q = 63.90, P is less than 0.05), but higher than that in control group (q = 108.63, P < 0.01). The expressions of TNFalpha and IL-1beta mRNA and proteins in liver were significantly decreased in BD+SB203580 group compared with the BD group (q = 55.11, 98.13, 61.03 and 50.85, respectively, P < 0.01), but higher than that in control group (q = 68.89, 37.22, 182.06 and 141.38, respectively, P < 0.01). SB203580 can suppress the expression of pro-inflammatory cytokines in the liver of brain dead rats through the inhibition of p38MAPK signaling pathway which may reduce the immunogenicity of donor livers.

Animals ; Brain Death ; metabolism ; Imidazoles ; pharmacology ; Inflammation ; Interleukin-1beta ; metabolism ; Liver ; drug effects ; metabolism ; pathology ; Male ; Pyridines ; pharmacology ; Rats ; Rats, Wistar ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism