Objective To study the simple preparation method and structure of nano-hydroxyapatite/collagen composite, to investigate new substitute of repairing bone for using in tissue engineering. Methods Porous nano-hydroxyapatite was made of Ca (OH)2 and H3PO4. Collagen was drawn from fresh adult bovine tendon. The two materials were prepared into biomembrane through the glutaraldehyde and freeze-drying. The crystallite phase, micro-morphology, structure, crystallite size of the composite were examined by XRD and scanning electronic microscop (SEM). Results The results showed that the composite structure was porous and consisted of nano-hydroxyapatite (10 nm ? 50 nm - 20 nm ? 80 nm) and collagen fiber. The crystallite phases and size of the composite was similar to that of natural bone. Conclusion The porous nano-hydroxyapatite /collagen composite is expected to be an ideal substitute of repairing bone.

OBJECTIVE:To optimize the content determination conditions of total alkaloids from Zhuang medicine Munronia delavayi,and to determine the content of total alkaloids in M. delavayi from different producing areas. METHODS:With the con-tent of total alkaloids as index,using solvent amount,ultrasonic time,ultrasonic extraction times and pH value of buffer as fac-tors,the content determination conditions of total alkaloids from Zhuang medicine M. delavayi were optimized by orthogonal test. Optimized content determination conditions were adopted to determine the content of total alkaloids in M. delavayi from 18 produc-ing areas in different harvest time. RESULTS:The optimum content determination conditions were as follows as the amount of sol-vent(CHCl3)20 ml,ultrasonic processing for 3 times,lasting for 15 min each time,pH value of buffer 4.5. The contents of total alkaloids in M. delavayi from 18 producing areas were between 0.6-11.98 mg/g,showing great difference. M. delavayi from Long-lin county and Tianlin county harvested in Oct. had the lowest content of total alkaloids. CONCLUSIONS:Optimized content deter-mination condition can be used for the content determination of total alkaloids in Zhuang medicine M. delavayi and quality control of it. The content determination of total alkaloids in M. delavayi is related to producing area and harvest time.

Objective To summarize the experience of surgical management of carotid tumors ( CBT ) and application of shunt between common and internal carotid artery intraoperatively. Methods Thirty patients of CBT (mean age:39. 2 ±2. 3 years old,10 male and 20 female, 15 in left, 14 in right and 1 in both sides) who underwent surgical resection, were retrospectively reviewed. The average size of CBT was 4.9 ±0.3 cm. The diagnosis was established by ultrasound, CT, MRI or carotid arteriography. 16 patients underwent surgical resection of CBT, 10 patients underwent additional ligation of external carotid artery, and 4 patients underwent additional intraoperative shunt between common and internal carotid artery. Results Surgical procedures were successfully performeded in all 30 patients with CBT. Intraoperative shunts were successfully used between common and internal carotid artery in 4 patients. The postoperative complications included hoarseness (15) , bucking (11), crooked tongue ( 17) , dyspnea (1), dysphagia(3). There was no hemiplegia and death. Conclusion Surgical resection is the choice of treatment of carotid body tumor. The application of intraoperative shunt between common and internal carotid artery in complicated Shamblin Ⅲ stage is safe and effective.

OBJECTIVETo explore the chemopreventive effect of curcumin on DMH induced colorectal carcinogenesis and the underlining mechanism.

METHODSTotally 40 Wistar rats were divided into the model group and the curcumin group by random digit table, 20 in each group. Meanwhile, a normal control group was set up (n =10). A colorectal cancer model was induced by subcutaneously injecting 20 mg/kg DMH. The tumor incidence and the inhibition rate were calculated. The effect of curcumin on the expression of peroxisome proliferator-activated receptor gamma (PPARγ) in rat colon mucosal tissues was observed using immunohistochemistry and Western blot. HT 29 cell line were cultured and divided into a control group, the curcumin + GW9662 (2-chloro-5-nitro-N-4-phenylbenzamide) intervention group, and the curcumin group. The inhibition of different concentrations curcumin on HT29 cell line was detected using MTT. The expression of curcumin on PPARy was also detected using Western blot.

RESULTSThe tumor incidence was 80. 00% (12/15 cases) in the model group, obviously higher than that of the curcumin group (58. 82%, 10/17 cases, P <0. 05). The inhibition rate of curcumin on DMH induced colorected carcinoma reached 26. 46%. Compared with the normal control group, the expression of PPARγ protein was significantly increased in the curcumin group and the model group (P <0. 01). Compared with the model group at the same time point, the expression of PPARy protein was significantly enhanced in the curcumin group (P <0. 05). MTT analysis showed that curcumin could inhibit the proliferation of in vitro HT 29 cells in dose and time dependent manners. The expression of PPARy protein was significantly increased in the GW9662 group and the curcumin group, showing statistical difference when compared with the normal control group (P <0. 01). Compared with the GW9662 group, the expression of PPARγ protein was significantly increased in the curcumin group (P <0. 01).

CONCLUSIONCurcumin could inhibit DMH-induced rat colorectal carcinogenesis and the growth of in vitro cultured HT 29 cell line, which might be achieved by activating PPARy signal transduction pathway.

Anilides ; Animals ; Carcinogenesis ; Colorectal Neoplasms ; drug therapy ; metabolism ; Curcumin ; pharmacology ; therapeutic use ; PPAR gamma ; metabolism ; Rats ; Rats, Wistar ; Signal Transduction

[Objective] Based on the resting state functional magnetic resonance imaging to investigate the abnormal features of the functional connectivity of resting brain neural network in the patients with primary insomnia,by using voxel-wise whole-brain finctional networks analysis of degree centrality (DC) for imaging evidence of neural mechanisms underlying primary insomnia.[Methods] The resting state fMRI were performed in 59 PI patients and 47 age,education,and sex-matched normal healthy subjects.Analysis of DC map changes between the two patient groups and the control group were performed by two sample t test.(threshold at P ＜ 0.05).[Results] Compared with the control group,the patients with PI showed significantly reduced DC value in bilateral medial frontal gyrus (MFG),bilateral anterior cingulate gyrus (ACG),and right insula;and increased DC value in right middle temporal gyrus (MTG),and left cuneus,(CUN),P ＜ 0.05.[Conclusion]Changes of DC value occurred in some region of brain in the P[patient groups when compared with the control group.It was indicated that DC,as a novel resting-state fMRI parameter in the voxel-wise whole-brain functional networks,might be an appealing alternative approach for further study on pathologic and neuropsychological states of PI.

Background Dyslipidemia is one of the major causes of age-related macular degeneration (AMD).At present,the study of the preventive and treating methods of A MD is still a hot spot.Objective The purpose of this study was to observe the effect of tonifying the spleen and promoting blood circulation on the retina and Bruch membrane in apolipoprotein E-deficient (ApoE-/-) mice with dyslipidemia.Methods Thirty-six ApoE-/-mice aged 2 months were randomly divided into the normal diet group,high fat diet group and medicine group.A diet with a higher content of fat was given for 5 consecutive months to the mice of the high fat diet group and medicine group,and in the last month,a concoction that tonifies the spleen and promotes blood circulation was gavagely administered in the medicine group,and an equivalent volumes of normal saline solution was administered in the same way in the normal diet group and high fat diet group.Total plasma cholesterol (TC),low density lipoprotein (LDL)and triglyceride (TG) were detected by (ELISA? Name of assay?) using the 7170 Hitachi automatic biochemical analyzer,and morphological changes of the retina and Bruch membrane were examined by light microscopy and transmission electron microscopy.The number of outer nuclear layer (ONL) cells,retinal pigment epithelial (RPE) cells and the thickness of the Bruch membrane were examined by semi-quantitative histopathology with the Mias 2000 Imaging Analyzer System.Results The concentrations of TC,LDL and TG were (6.47 ±0.49) mmol/L,(1.46 ±0.10)mmol/L and (0.62 ±0.21) mmol/L,respectively,in 7-month-old mice of the medicine group,showing a significant reduction in comparison with (10.53 ±0.30) mmol/L,(1.90±0.13) mmol/L,(1.15±0.29) mmol/L of the high fat diet group,and (9.63 ± 0.18) mmol/L,(1.12 ± 0.15) mmol/L,(0.88 ± 0.21) mmol/L in the normal diet group (P＜0.05-0.01).The disorder and atrophy of ONL and RPE cells,divergence of fiber of the Bruch membranes were found in both the high fat diet group and normal diet control group under the light microscope,and drusen formed in some of the mice in the high fat diet group.However,ONL and RPE were well organized in the medicine group.The cell numbers in the ONL and RPE layer in the 7-month-old mice were (23 124.00±755.18) and (10.75±0.59),respectively,in the medicine group,(19 107.00 ± 1436.82) and (8.55 ± 1.11),respectively,in the high fat diet group,(21 663.00± 1073.27) and (9.75 ±0.58),respectively,in the normal diet group,with significant differences among them (P＜0.05-0.001).Thickness of the Bruch membrane in the medicine group extensively reduced in high fat diet group and normal diet control group (P＜0.01).The ultrastructures of the RPE and Bruch membrane were much more improved in the mdedicine group.Conclusions Tonifying the spleen and promoting blood circulation can attenuate hyperlipemia in ApoE-/-mouse;furthermore,it lessens the pathological abnormalities in the ONL,RPE and Bruch membrane.

Objective To explore the effect of dexmedetomidine combined with ropivacaine for continuous lumbar plexus block on the quality of early recovery in patients undergoing total hip ar-throplasty.Methods Sixty patients (35 males and 25 females)with ASA physical status Ⅱ or Ⅲ, aged 65-84 years,undergoing total hip replacement were randomly assigned to dexmedetomidine com-bined with ropivacaine group (group D)or ropivacaine group (group C).All patients received ropiva-caine which was administered via continuous lumbar plexus block as patient-controlled analgesia (PCA)after surgery.The PCA were programmed with a background infusion 8 ml/h of ropivacaine, bolus dose was 4 m1 and the block time was 30 min;the patients in group C received 0.2% ropiva-caine,and the patients in group D received 0.1% ropivaciane combined with 1 μg/ml of dexmedeto-midine by the way of intravenous infusion.In addition,all patients received another patient-controlled intravenous analgesia (PCIA)with 1 mg/ml of morphine for relieving the explosive pain.The PCIA was programmed with a lobus dose of morphine 1 mg without background dose,the block time was 5 min.Consumption of morphine and visual analog scale (VAS)score,muscle strength and maximum flexion and abduction of hip joint were recorded at the time points of 6,12,24 and 48 h after opera-tion.The side-effect reactions such as nausea,vomiting,drowsiness and itching were recorded.The sleep quality was assessed with Pittsburgh sleep quality index (PSQI)on day 1 before operation,day 1 and day 7 after operation.Results The consumption of morphine in group D was significantly fewer than group C (P <0.05).Compared with group C,muscle strength and maximum flexion and abduc-tion of hip joint in group D were increased at 6,12,24 and 48 h after operation (P <0.05 ).PSQI scores were decreased on the day 1 and day 7 after operation.The incidence of delirium,nausea and vomiting,drowsiness and itching in group D were decreased (P <0.05).Conclusion 0.1% ropiva-ciane continuous lumbar plexus block combined with 1 μg/ml of dexmedetomidine by the way of in-travenous infusion could provide satisfied analgesia for the operation of total hip arthroplasty;the sleep quality is improved,the functional exercise of hip joint is promoted.

Objective To evaluate the effectiveness of arthroscopically assisted percutaneous reduction and internal fixation with eannulated screws.Methods The fracture was reduced by closed manipulation or percutaneous leverage force by using the Kirschner wire.Then the patella was temporarily fixed by using a large size towel clamp or Kirschner wires.Under the guidance of knee arthroscopy,a micro-incision was made at the size of cannulated screw placement,the pilot holes were drilled at a proper depth,and the thread was configured.Two titanium screws were inserted parallelly.Results Following-up chekups for 4～24 months in 18 cases showed a satisfactory recovery of knee functions.According to the Bostman' standard,excellent effects were obtained in 16 cases and good effects in 2 cases.Conclusion Treatment of patellar fractures by percutaneous cannulated screw fixation under arrhroscope of- fered advantages of minimal invasion,accurate reduction,reliable fixation,and excellent recovery of joint functions.

Constructing prokaryotic expression vector pKY-RMBAY by gene recombination and research its optimizing productive conditions.By PCR technology synthesizing the gene of the RMBAY with preference codon of E.coli and the RMBAY gene was inserted into high efficiency expression vector pKYB-MCS.Expressed fusion proteins in E.coli ER2566 were purified with Chitin-Beads column.Fusion proteins binding on Chitin-Beads was cut on N-terminus of intein due to the induction of ?-mercaptoethanol and the target peptide RMBAY was released.The RMBAY was identified by mass spectrum.Experiment results showed RMBAY can be high efficiently expressed in E.coli ER2566,with optimizing productive conditions the yield of the RMBAY may be 6.7mg/L fermentation product and its purity is greater than 98%.The molecular weight of RMBAY is 3.887 kDa by mass spectrum and that accords with its theory value.

Background There is no effective method to regenerate the optic nerve after injury. It has been recently reported that α-crystallin could promote the survive rate and axon regeneration of retinal ganglion cells (RGCs) effectively. However,the molecular mechanism is not clear. Objective This study was to identify the site of RGCs where the exogenous α-crystallin bind to. Methods RGCs was isolated from retinas of two 2-day-old Long Evans rats and primarily cultured. The positive rate of the RGCs was assessed by counting the number of positive cells for fluorescently-labeled thy1. 1 and cy3 under the fluorescence microscope. The biotinylated exogenous α-crystallin was evaluated by direct coloration and the activity of molecular chaperones was measured by insulin test.After identifying the success of biotinylation along with the activity of molecular chaperones,biotinylated α-crystallin was co-incubated with RGCs and the cells then were reacted to fluorescently labeled avidin for the observation of binding site of exogenous α-crystallin under the laser confocal microscope. Results RGCs of 94% were survived through primary culture. The coloration of biotinylated α-crystallin labeled by the direct coloration method was more intensive, and the value of A450 descended as the decrease of biotinylated α-crystallin concentration,indicating that the α-crystallin was biotinylated successfully. The activity of molecular chaperones of biotinylated α-crystallin was significantly strong but no significant change after being biotinylated after co-incubation of RGCs with biotinylated α-crystallin. Laser confocal microscope examination revealed that co-incubated RGCs with biotinylated α-crystallin showed the red fluorescence on membrane and axon of RGCs rather than cytoplasm and nucleus. The absent response was seen in the control group. Conclusion Exogenous α-crystallin can specifically combine with the membrane of RGCs to play the biological function,but its binding mode and mechanism need further study.