Objective To investigate the antimicrobial properties of human amniotic homogenate supernatant (HAHS) in order to found a theoretical base for the use of human amniotic membrane in ophthalmological field. MethodsFresh human amniotic membranes were used to make HAHS and acellular amniotic membranes. Then, we observed their antimicrobial effects and antimicrobial spectrums, compared the antimicrobial capacity with 10 commonly used antibiotics in eyes, and investigated the effects of time, temperature and pH value on the antimicrobial capacity. Finally, transmission electron microscopy (TEM) was used to explore the possible targeting site of the antibiosis. ResultsHuman amnion membrane contained antimicrobial components locating in its epithelial cells. HAHS had a broad antimicrobial spectrum and was steady in nature. Its antimicrobial capacity was stronger than those of sulfasulfonamide, chloromycetin and cefuroxime sodium. TEM indicated that the antimicrobial effect were exerted through plasma membrane of microorganism. ConclusionHAHS can be an effective and convenient treatment for ocular surface infectious diseases. Traditional amnion transplantation should employ fresh human amniotic membranes containing complete epithelial lamina to reconstruct the ocular surface.

This study aimed to assess the specificity of auricular acupoints in responding to changes of qi and blood which, in traditional Chinese medicine, are considered as the fundamental substances in the human body for sustaining normal vital activity.

In this study, to analyze the influence of the brightness value of the supranasal point and the apex nasi on their dominant wavelength and excitation purity according to the spectrocolorimetry data of the supranasal point and the apex nasi in healthy adults that were collected based on optical spectrum colorimetry.

Objective To express and purify recombinant and biologically active Clostridium difficile toxin B (rTcdB). Methods The genes of TcdB were amplified by polymerase chain reaction (PCR) using chromosomal DNA from a toxigenic strain, and cloned into a shuttle vector pHis1522.The sequences of TcdB genes in the vector were verified by DNA sequencing. The construction was transformed into Bacillus megaterium protoplasts and the protein expression was driven by a xylose promoter. The purified protein was tested for biological activity. Results rTcdB was successfully purified from bacterial crude extracts. Approximately 5-10 mg of highly purified recombinant toxin was obtained from one liter of bacterial culture. The expressed rTcdB had molecular mass similar to the native toxin, and its biological activity was proved to be similar to its native counterpart after an extensive examination. Conclusion rTcdB with biological activities is successfully expressed in Bacillus megaterium.

Objective To investigate the effects of chondroitin sulfate proteoglycans (CSPGs) degradation induced by chondroitinase ABC (ChABC) on the rat flash visual evoked potential (F-VEP) around the end of the critical period of visual development.Methods A total of 30 Long Evans rats were randomly divided into 3 groups according to their age of 14,21 and 35 d.Then in each group,5 animals served as CSPGs degradation model induced by ChABC microinjection into 5 sites of the rat cortex [750 nL (48 U/ml) for 1 site,for 2 times at a 3-day interval],and the other 5 rats served as normal control and received normal saline microinjection.Immunofluorescence microscopy and laser scanning confocal microscopy were used to determine the establishment of animal model.F-VEPs were evaluated in 7 d after first microinjection.Results In the normal rats,the latency of F-VEP main wave became shorter with the increase of their age,the number was [(63.5?10.1),(50.8?6.4),(44.9?6.3) ms,P

To study the effect of Buyang Huanwu Decoction (BHD) on neural cell apoptosis after cerebral ischemia in gerbils and its mechanism, gerbil model with cerebral ischemia was established by clamping of bilateral carotid arteries and then reperfusion. Neural cell apoptosis of hippocampal gyrus slides was observed under electron microscope 120 hours after reperfusion. The activities of Na + -K + ATPase , Ca 2+ ATPase , Mg 2+ ATPase and NOS, and the contents of lactic acid, NO and amino acid in cerebral tissues were detected 120 hours after reperfusion. The results showed that BHD can reduce the incidence of neural cell apoptosis of hippocampal gyrus, prevent the decrease of Na + -K + ATPase and Ca 2+ ATPase activities, increase NO content and NOS activity, and reduce cerebral glucose content. It is indicated that BHD can counteract neural cell apoptosis after cerebral ischemia in gerbils and its mechanism may be related to the improvement of cerebral energy metabolism, regulation of NO synthesis and antagonism of toxic effect of excitatory amino acid.

During placental development, oxygen environment is not only critical for trophoblasts migration and invasion, but also fundamental for appropriate placental perfusion. Cysteine-rich 61 (Cyr61, CCN1) was expressed in the extravillous trophoblasts (EVTs) and decreased in preeclampsia. Its regulatory properties in human first-trimester extravillous trophoblast cell line (TEV-1 cells) upon a low oxygen tension were investigated. The present study examined functional changes involved in adaptation to hypoxia of the TEV-1 cells, using cobalt chloride (CoCl(2)) as hypoxic mimic. It was found that hypoxia inhibited growth of TEV-1 cells and induced the increase of cell apoptosis (P<0.05). The Cyr61 expression in human EVTs was transcriptionally induced by CoCl(2). Inappropriate EVTs apoptosis has been implicated in the failure of trophoblasts to fully invade and modify the uterine environment and Cyr61 down-regulation, potentially leading to preeclampsia.

OBJECTIVE To type the genes of plasmid DNA in 54 clinical Klebsiella pneumoniae isolates producing extended -spectrum beta-lactamases (SHV) by denaturing high-performance liquid chromatography (DHPLc) and evaluate their sensitivity and specificity, and explore a rapid and convenient method for detecting the antibiotic resistance of K. pneumoniae. METHODS Plasmid DNA from each extended-spectrum beta-lactamase (SHV) producing strain was subjected to PCR amplification. After we performed DNA sequencing of these amplicons and identification of mutation and their genotype, DHPLc was undertaken to investigate whether its results correlate the distinctive chromatogram with each genotype. RESULTS All the strains were found abnormal elution peaks (two or three peaks) which were different from each other. The result of DNA sequencing demonstrated that all the strains had DNA mutation in comparison with SHV-1. Moreover, DHPLc could produce specific peak patterns that correlate with genotype. CONCLUSIONS The sensitivity of DHPLc is 100% in this study. And each genotype is corresponded to specific peak pattern. So we can use DHPLc technique to type the genes of plasmid DNA in K. pneumoniae and detect mutations rapidly. DHPLc not only has high accuracy , but also is a convenient and rapid technique for the detection of mutation in the bacterial genome. It has a great potential clinical value.

Objective To identify and quantitate the conjunctival aerobic bacterial flora from healthy people of 3 age groups in Chongqing city.Methods Samples taken with moist swabs from the lower fornix of both eyes of 33 children(9.85?0.57 years old),31 youths(21.23?0.88 years old)and 30 elders(72.97?4.94 years old)were given bacterial cultures.Positive ones were identified and colony forming units(CFUs)were counted.Bacterial quantity was expressed as the number of CFUs divided by the weight of each sample measured before and after collection.Results The elders showed a higher isolation frequency(93.33%)than children(45.45%)and youths(50.00%)which had statistically difference(P

0.05).Conclusion The rat corneal neovascularization induced by alkali cauterization can be significantly inhibited by local application of 0.025%,0.1% Nordy eye drops that have no toxicity and adverse effect.