[Objective] Subtractive cDNA libraries from the leaves of D. candidum were constructed to supply evidence for the clone of dendrobine-biosynthesis-associated genes. [ Methods ] Suppression subtractive hybridization ( SSH) technology was used. With the cDNA of perennial leaf as the tester and that of annual leaf as the driver, forward and reverse hybridization was performed. The two obtained subtracted cDNA fragments were cloned into PointTMXa-1 plasmid vectors, and then the vectors were transformed into E. Coli JM109. The inserted fragments were amplified by PCR and then identified. [Results] The subtractive cDNA libraries related with dendrobine-biosynthesis-associated genes were successfully constructed. The forward and reverse subtractive libraries included 560 and 220 clones respectively. Detected by PCR, the length of the inserted fragments was 550 bp on average. [Conclusion] The constructed subtractive libraries are suitable for further study on the functional genes associated with dendrobine biosynthesis of D. candidum.

[Objective] cDNA expression libraries from the leaves of Dendrobium candidum Wall. ex lindl. were constructed to supply evidence for the clones of the full-length genes involved in the biosynthesis of bioactive compounds. [Methods] Total RNA was isolated and purified from tender leaves of 4-year-growing Dendrobium candidum by using Trizol single-step method. cDNA was synthesized by long distance polymerase chain reaction (LD-PCR) and then was connected to ?TripIEX2. After that, the recombinant bacteriophages were packaged and cDNA library of Dendrobium candidum was constructed. The titer of cDNA library was expressed as the number of phage formation unit (pfu) per milliliter and the inserted fragment was identified by PCR amphfication. [Results] cDNA expression libraries from the leaves of Dendrobium candidum were constructed successfully. The titer of the original library was 4.3?105 pfu/mL and 3.1?105 clones in total, with a recombinant rate of 97.6%. The amplified titer was 6.8?109pfu/mL. PCR amplification suggested that the inserted cDNA fragments ranged from 0.5 to 2.0 kb, mostly from 1.2 to 1.7 kb. [Conclusion] The constructed Dendrobium candidum cDNA library has a higher titer, a higher recombinant percentage and larger inserted fragments.

OBJECTIVE:To establish a HPLC method for determination of the content of baicalin in Bifukang oral liq?uid.METHODS:The ODS C 18 column was used with methanol-water-glacial acetic acid(45∶55∶1.5)as mobile phase,and de?tective wavelength was274nm,the flow rate was1ml/min.RESULTS:Within the range of0.215?g～4.3?g,baicalin presented a fine linear relationship between amount and peak area(r=0.9995).The average recovery was99.44%(RSD=0.79%,n=5). CONCLUSION:This HPLC method is convenient,rapid and accurate,and can be used for quality control in the production of Bifukang oral liquid.

Object To explore the protoplast fusion of Dendrobium candidum Wall ex. Lindl. and Gynostemma pentaphyllum. (Thunb) Makino. Methods The two kinds of protoplasts were fused by PEG method, and then cultured in the modified liquid medium containing 2 mg/L BA and 1 mg/L NAA. Results High yield, viability, and pure mesophyll protoplasts were isolated from D. candidum and G. pentaphyllum. Conclusion The first cell division occurres within three days after fusion. And some of the cells are divided three times.

Objective To explore the effects of interleukin1?(IL-1?) and matrix metalloproteinase-9(MMP-9) in middle ear cholesteatoma and to explore the correlation between their expression and the ability of destruction of cholesteatoma.Methods IL-1? and MMP-9 were determined immunohistochemically in the specimens from 21 cases cholesteatomas and 10 external ear skin of patients with cholesteatoma.Results The positive expression of IL-1? and MMP-9 in cholesteatoma epithelialium were increased greatly than that in external epidermis(P

Objective To evaluate effects of steep meridian clear corneal incisions on corneal anterior surface and total astigmatism in phacoemulsification surgery . Methods Clinical data of 56 cases of phacoemulsification surgery with steep meridian clear corneal incisions from August to December 2015 were analyzed in this retrospective study .Corneal anterior surface and total astigmatism were measured with the Pentacam 3D anterior segment measurement and analysis system at the time before surgery and 1 month and 3 months after surgery .The polar coordinate analysis was used to evaluate effects of steep meridian incisions on corneal astigmatism . Results All the operations were accomplished successfully with smooth recovery .The corneal anterior surface and total astigmatism reduced significantly after 1 month and 3 months in astigmatic polar value AKP ( +0) (P<0.05), but no significant difference was found in AKP(+45) (P>0.05).No significant differences were detected between both corneal anterior surface and total astigmatism in AKP(+0) or AKP(+45) between 1 month and 3 months after surgery. Conclusion Steep meridian incision performed on the preoperative steeper meridian of keratometric astigmatism may reduce corneal total astigmatism .

Objective To discuss the efficacy of photoselective vaporization of the prostate(PVP) for patients with benign prostatic hyperplasia(BPH).Methods From July 2006 to August 2007,110 patients with BPH received PVP in our hospital.The safety of the procedure,Pre- and postoperative Qmax,and IPSS of the cases were recorded and analyzed.Results The mean operation time was(51.2?36.3) minutes(ranged from 15 to 180).In the patients,23 cases had a prostate weighed ≥100 g,9 of them received TURP during the procedure.89 patients underwent bladder irrigation for 15 to 48 hours(mean,36 hours).After the operation,urinary catheter was left indwelling in all the patients except for 6(

BACKGROUND:Pulsed laser deposition synthesis technology has been used to prepare new nano-hydroxyapatite thin film coating by colagen deposition on artificial mechanical heart valve. OBJECTIVE: To investigate the toxicity of new nano-hydroxyapatite thin film on human umbilical vein endothelial cels. METHODS: Human umbilical vein endothelial cels were cultured with nano-hydroxyapatite film room-temperature leaching solution, nano-hydroxyapatite film high-temperature leaching solution, high-density polyethylene and phenol solution. Within 72 hours, cel growth was observed under the inverted phase contrast microscope. At 7 days after culture, cel proliferation and toxicity grading were detected using Cel Counting Kit-8. RESULTS AND CONCLUSION:At 24 hours after culture, cels grew wel, showed fusiform shape, and had strongrefraction in the nano-hydroxyapatite film room-temperature leaching solution, nano-hydroxyapatite film high-temperature leaching solution, high-density polyethylene groups, and no significant differences in cel morphology and number were detected among above groups. Cels in the phenol solution group were suspended, round, pyknotic and dead. At 48 hours, except phenol solution group, cel number increased significantly, and cel grew densely in other three groups. At 72 hours, cels grew strongly, and the gap became smal obviously. Within 7 days after culture, cel proliferation activity was not significant in the nano-hydroxyapatite film room-temperature leaching solution, nano-hydroxyapatite film high-temperature leaching solution, and high-density polyethylene groups, which was significantly higher than in the phenol solution group (P < 0.05). The toxicity of nano-hydroxyapatite film graded 0 to 1. These results suggested that nano-hydroxyapatite artificial mechanical heart valve has good histocompatibility, but no toxicity.

BACKGROUND:A new type of nano-hydroxyapatite artificial mechanical heart valve has been developed using pulsed laser deposition technology at the Department of Materials, Hefei University and Anhui Institute of Optics and Fine Mechanics, Chinese Academy of Sciences, China. OBJECTIVE:To investigate the compatibility of nano-hydroxyapatite artificial mechanical heart valve with human umbilical vein endothelial cels. METHODS:Human umbilical vein endothelial cels were in vitroisolated, cultured and passaged to the 2-4 generations, and then the cel suspension was inoculated onto the nano-hydroxyapatite artificial mechanical heart valve. After 3, 7, 12 days of culture, the cel growth on the artificial mechanical heart valve was observed under scanning electron microscope. In addition, the human umbilical vein endothelial cels were respectively cultured in room-temperature and high-temperature extract liquids of nano-hydroxyapatite artificial mechanical heart valve, high-density polyethylene and phenol solution extracts for 72 hours, and then, the proliferation of cels was detected by MTT method. RESULTS AND CONCLUSION:Under the scanning electron microscope, the human umbilical vein endothelial cels were fusiform- or polygon-shaped with protuberances adhered to the artificial mechanical heart value at 3 days of culture; the cels were stretched thoroughly and fused at 7 days of culture; and the cels were confluent to pieces that tightly overlaid the heart valve surface and the extracelular matrix was formed localy at 21 days of culture. Results from MTT test displayed that the nano-hydroxyapatite artificial mechanical heart valve had no cytotoxicity to the human umbilical vein endothelial cels, indicating a good cytocompatibility.