At present, filtration surgery remains an important treatment of glaucoma, and filtering bleb fibrosis is the main cause for treatment failure. Filtering bleb fibrosis is a common fiber hyperplastic disease, and it relates to the activation and proliferation of fibroblasts and the excessive production of extracellular matrix ( ECM) such as collagen protein. The most frequently-used drugs for filtering bleb fibrosis in clinic are 5-fluoro-2,4 (1h, 3h) pyrimidinedione ( 5 - Fu ) and mitomycin ( MMC ) . Although they are effective in some degree, they also have some serious side effects which restrict their clinical use. Triptolide ( TPL) is a major active component of the medicinal plant, tripterygium wilfordii hook. f. ( TWHF) . TPL has multiple pharmacological activities including immunosuppressive, anti- inflammatory, anti- cancer and anti-fertility activity. Reviewing related literatures published in recent ten years, we confirmed that TPL seemed to possess a pharmacological activity in treating filtering bleb fibrosis. Since it has three major functions:1. inhibit the activation and proliferation of fibroblasts and the excessive production of collagen protein; 2. alleviate the inflammatory reaction after surgical wound to suppress fibrous scar formation; 3. TPL has a protective effect on retinal ganglion cells ( RGCs ) . We further find that TPL's anti-fibrosis activity mainly results from that it inhibits TGF-β/Smad,NF-κB and PI3K/AKT signal transduction pathway. This comprehensive analysis about the feasibility of Triptolide's role in treating filtering bleb fibrosis after the filtration surgery of glaucoma can help us develop new drugs for filtering bleb fibrosis and exploit TPL's clinical value on some level.

Objective To investigate the induction mechanism of MT ⅢmRNA in the brain after cerebral ischemia.Methods The forebrain ischemia reperfusion model was established in rats. The changes of the expression of MT Ⅲ mRNA in hippocampus after forebrain ischemia reperfusion were observed by in situ hybridization method. The changes of free Zn 2+ in hippocampus after forebrain ischemia reperfusion was examined using Zn 2+ specific fluorescent probe(TSQ). The Zn 2+ chelator (CaEDTA) was injected into the lateral ventricles for determining influences of Zn 2+ on the MT Ⅲ mRNA expression and the neuronal damage after forebrain ischemia/reperfusion.Results (1) The expression of MT ⅢmRNA in hippocampus increased gradually after cerebral ischemia and reached the peak in 96 hours after reperfusion. Seven days after reperfusion the expression of MT ⅢmRNA reduced to the normal level. (2) Zn 2+ fluorescence in the hilus of dentate gyrus, CA 3 region and the stratum radiatum and stratum oriens of CA 1 decreased slightly at 48 hours after reperfusion. From 72 to 96 hours after reperfusion, the fluorescence returned to normal, but some new fluorescence dots appeared in pyramidal neurons of CA 1 and the hilus of dentate gyrus increased gradually. Seven days after reperfusion, the fluorescence returned to normal. (3) The cell membrane impermeable Zn 2+ chelator could reduce the intracellular concentration of free Zn 2+ and the expression of MT Ⅲ mRNA.Conclusion The expression of MT Ⅲ mRNA can be induced by the increase in the concentration of intracellular free Zn 2+ after forebrain ischemia/reperfusion.

Animals ; Brain Ischemia ; metabolism ; pathology ; Male ; Metallothionein ; metabolism ; Nerve Tissue Proteins ; metabolism ; Neurons ; pathology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism ; pathology

Objective To explore the clinical value of wave intensity (WI) in evaluating early changes of cardiovascular function in patients with type 2 diabetes mellitus (T2DM).Methods Thirty T2DM patients without atherosclerosis by conventional ultrasonography were enrolled as case group,while thirty healthy volunteers were enrolled as control group.Two groups were well-matched in age and sex.Both of the case and control groups were taken WI test of carotid artery.The parameters of WI examinations in two groups were measured and compared.Results Compared to the control group,values of arterial stiffness index (β),elasticity modulus (Ep),pulse wave transit velocity (PWVβ),and decelerating wave intensity (W2) were significantly higher in T2DM group (P ＜0.05).Arterial compliance (AC) value was significantly reduced in T2DM group (P ＜0.05).Accelerating wave intensity (W1) value was higher in T2DM group without significant statistical differences (P ＞ 0.05).Conclusions WI technique which can early assess the changes of left ventricular function and vascular elasticity in T2DM patients has important clinical value.

Objective To investigate the role of apoptosis and the regulating role of receptor interacting protein 1(RIP1) in acute pancreatitis .Methods Thirty C57 mice were divided into three groups :control group ,acute edematous pancreatitis (AEP) group and acute necrotizing pancreatitis (ANP) group .The AEP group was continuously injected by cerulein 50 μg/kg for 13 times ,the ANP group was continuously injected by cerulein 50μg/kg for 13 times and lipopolysaccharide 15 mg/kg once;the con‐trol group was injected by the same volume of normal saline for 7 times .The acinar cell apoptosis was observed by the terminal de‐oxynucleotidyl transferase‐mediated deoxyuridine triphosphate nick‐end labeling (TUNEL) assay .The RIP1 mRNA expression was measured by real time fluorescence PCR .The expression of RIP1 protein was detected by Western blotting .Results The mouse models of AEP and ANP were established successfully .Compared with the control group ,acinar cell apoptosis existed in both AEP and ANP model groups ,moreover compared with the AEP group ,apoptosis in the ANP group were decreased ,the differences were statistically significant(P<0 .05) .Compared with the control group ,the expression of RIP1 mRNA and protein in the AEP group was increased ,while which in the ANP group were decreased ,the differences were statistically significant(P<0 .05) .Conclusion RIP1 participate in the pathogenesis of acute pancreatitis ,which may associate with acinar cell apoptosis .

Glaucoma is the first leading cause of irreversible blindness and the second leading cause of blindness worldwide. Numerous studies have shown that elevated intraocular pressure ( IOP ) is one of the major risk factors for the development and progression of glaucomatous optic nerve damage. However, there have been 50% of primary open-angle glaucoma ( POAG ) patients with typical glaucomatous optic neuropathy in whom the IOP measurements have always been in the normal range, and some patients develop typical glaucomatous optic neuropathy with the well controlled IOP. These phenomena cannot be explained by the theory of high intraocular pressure. The pathogenesis of glaucomatous optic nerve damage in these patients with normal IOP needs to be further discussed. Numerous studies at home and abroad have shown that: 1. the surrounding anatomy of the optic nerve including the IOP, the anatomy and biomechanics of the lamina cribrosa and retrobulbar orbital cerebrospinal fluid pressure may be of importance for the pathogenesis of the POAG;2. patients with normal tension glaucoma had significantly lower cerebrospinal fluid pressure and a higher trans - lamina cribrosa pressure difference compared with normal subjects; 3. patients with ocular hypertension had significantly higher cerebrospinal fluid pressure, however, there is no difference in trans -lamina cribrosa pressure compared with normal subjects. Based on the above research, now we make a review about the research advance of the relation between intracranial pressure and glaucoma optic nerve damage and the available measurements about noninvasive intracranial pressure in clinical in this paper.

Objective To study on algicidal bioactive substances from mangrove bacteria and its algicidal effect on red tide algae Alexandriam tamarense.Methods A red algicidal bioactive substance in Guangxi mangrove bacteria was isolated and purified by Silica-gel Column Chromatography,and its structure was determined on the basis of UV,IR,1H NMR,13C NMR and HREIMS spectroscopic analyses and comparison with the literatures.Its algicidal affect was primarily studied in this paper.Results and Conelusion The results showed that the red algicidal bioactive substance was prodigiosin,which had an inhibitory effect on the cells growth of Alexandrium tamarense,and the inhibitory effect increased with the increase of prodigiosin concentration.The prodigiosin stimulated the production of MDA and reduced the content of sulfhydryl group.In addition,plasma membrane permeability of leaked cell solution was increased.These indicated the extinguishing mechanism: the cell membrane was damaged followed by the inhibition of the cell growth.

BACKGROUND:Xenogeneic bone has a natural porous structure that is similar to human bone. In the treatment of bone defects, the porous structure is helpful to guide bone regeneration, but different degrees of immune responses wil be caused during the implantation process. OBJECTIVE:To prepare a freeze-dried antigen-extracted sheep cancelous bone scaffold and to evaluate its biocompatibility. METHODS: The sheep vertebral cancelous bone was colected to prepare two kinds of antigen-extracted heterologous bone scaffolds that were treated with chemical methods as chemical group and treated with chemical methods+cryopreservation at a-80℃ refrigerator for 4 weeks+drying in vacuum apparatus+60 RESULTS AND CONCLUSION:Freeze-dried bone had no cytotoxicity, no acute toxicity and heat reaction, and was negative for the intracutaneous stimulation test. The scaffold in the chemical group had cytotoxicity and mild irradiation as freeze-dried bone group. (1)Cytotoxicity test: Bone marrow mesenchymal stem cels isolated from sheep were cultured in extracts of the chemical group, free-dried bone group and Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12. (2) Heat reaction and acute toxicity tests: Extracts from the chemical group, freeze-dried bone group and normal saline were respectively injected into the ear vein of rabbits. (3) Intracutaneous stimulation test: Extracts from the chemical group, freeze-dried bone group and normal saline were respectively injected subcutaneously into the back of rabbits. Co acute toxicity reaction, sent heat source and had mild irritation. Results show that after freeze drying processing, the sheep vertebral cancelous bone has good biocompatibility, can meet the requirements of bone tissue engineering, but the bone that through chemical processing exhibits a relatively poor biocompatibility that cannot achieve the safety standard of biological scaffold materials.

Objective To establish a sensitive,specific,simple and high-throughput method for detection of the epidermal growth factor receptor (EGFR) mutation in the plasma samples of patients with non-small cell lung cancer (NSCLS) by the use of mutant-enriched liquid chip (MEL) assay.Methods The specific probes for the EGFR exon19 E746-750 deletion,exon 21 L858R mutation and wild-type sequence were designed and coupled to the microspheres coding with different fluorescent dye.The probe coupling efficiency was verified by crossing hybridization test with biotin-labeled reverse sequence.A blood-based MEL approach which integrates a sensitive mutant-enriched PCR and quantitative high throughput liquid chip assay for assessment of EGFR mutations was developed.The sensitivity and specificity of MEL was further evaluated using the mixture with different copy numbers of mutant and wild-type plasmids as template.The mutations of exon 19 and 21 of EGFR gene in plasma samples from 201 patients with stage ⅢB or Ⅳ NSCLC who enrolled in the First Affiliated Hospital of Guangzhou Medical College from September 2008 to April 2010 were analyzed by both the MEL and the mutant-enriched PCR assay.The result comparison was made between direct sequencing and MEL in 50 cases whose EGFR gene type had been tested by MEL and mutantenriched PCR.The correlation of EGFR gene mutation and the response to the Geftinib treatment was analyzed in 16 patients with lung adenocarcinoma as well.Results The probes were successfully coupled to the microspheres encoding with different fluorescent dye,and could be specifically recognized by the corresponding target sequence.The MEL was capable of detecting as few as 10 copies of EGFR mutants (sensitivity was 0.1％).Among the enrolled 201 cases of advanced NSCLC,the detection rate of the EGFR exon19 E746-750 del and exon 21 L858R was 55.7％ (112/201) by MEL assay.Conpared with mutantenriched PCR[58.2％ (117/201)],the coincidence rate was 97.5％ (196/201).There was no statistically significant difference between the results of mutant-enriched PCR and MEL (x2 =3.20,P ＞ 0.05).The mutations detection rate was 22.0％ (11/50) by directing sequencing,which was significantly lower than by MEL[50.0％ (25/50),x2 =12.07,P ＜ 0.05].Among the 16 patients treated with Gefitinib,9 cases who had EGFR mutation showed a higher response rate(P =0.041)and prolonged progression-free survival (x2 =6.76,P =0.009) after the treatment compared to those 7 who without EGFR mutation.Conclusions A new method of MEL with accuracy,specificity,fast and high-throughput is established for the detection of EGFR 19 E746-750 deletion and exon 21 L858R mutations in plasma from advanced NSCLC patients.It has the ability to provide the most direct and valuable guidance for clinicians to make decision on EGFR tyrosine kinase inhibitors therapy in the advanced NSCLC paticnts.

BACKGROUND:Xenogeneic bone structure and biological characteristics are similar to human bone tissue, and the xenogeneic bone has a decreased antigenicity after physicochemical treatment, with a natural porous structure and rich source, and can be kept for a long time, which is considered to be an effective way to solve the shortage of the autogenous bone and al ograft bone.
OBJECTIVE:To compare the physical and chemical properties of xenogenic bone materials prepared by two different methods.
METHODS:Sheep cancellous bone treated with chemical method was placed into the muffle furnace at 1 000 ℃for 2 hours to prepare calcined bone. Another cancellous bone was placed into an 80 ℃ refrigerator for 4 weeks and then placed into a vacuum instrument to prepare freeze-dried bone. Cancellous bone rinsed with ultra-pure water served as controls.
RESULTS AND CONCLUSION: Three groups of samples retained three-dimensional porous structure which similar with human bone tissue under microscopic observation. The framework was stil intact, with a smal pore of 55-650μm and high porosity of 65%-80%. For the calcined bone, the toughness was decreased and the brittleness increased significantly, but the freeze-dried bone had a little changes in the mechanical properties. Through diffraction analysis, hydroxyapatite was the main composition of the three groups. However, there was a smal amount ofβ-tricalcium phosphate in the calcined bone. Spectrum analysis confirmed that calcium and phosphorus content in these three groups were al close to the human body. The results suggest the cancellous bone treated with these two methods is similar to human bone structure, and the major elements are close to the body. In addition, the cancellous bone after processing has enough smal pore and higher porosity. However, calcination process has a more influence on the mechanical property of scaffold materials, and the freeze-dried bone has a little change but the antigen cannot be completely removed that can reach the basic requirements.