Nerve growth factor is one of the most important factors playing an important role in regulating the growth, development and survival of the neuron. The purified NGF from human placenta has been reported, the tissue from which can be isolated the NGF is very limited. It is important for basic research and clinic application to expression hNGF by genetic engineering. By polymerase chain reaction,gene fragment encoding the mature part of ?-NGF was amplified using the DNA of human placenta as template. The fragment was sequenced and inserted into expression vector pET-15b, and the recombinant expression vector pET15b-NGF was transformed into E.coli BL21(DE3)pLysS. After inducing with IPTG the NGF was higher expressed up to 25% of the total cell proteins. The expression product was purified with metal chelate affinity chromatography on Ni-IDA agarose under denaturing condition. The purity of rNGF was higher than 90% and yield of rNGF was 4.56mg/L expressing bacteria. SDS-PAGE revealed the NGF expression product had a Mr 16kDa. Western-blot displayed the recombinant product had strong immunological activity with rabbit anti-human ?-NGF polyclonic antibodies. The expression products were dealed with solubilizing inclusion bodies and refolding protein. The test of nerve fiber growth of chicken embryo DRG neurons displayed rNGF had biological activity.

OBJECTIVETo develop a new method for the determination of cholesteryl palmitate in Oviductus Ranae.

METHODA HPLC method was set up, using Zorbax Silica column and cyclohexane-diethyl ether (40:1) as mobile phase with a flow rate of 1.0 mL x min(-1), and the UV detection wavelength was 203 nm.

RESULTThe calibration curve was linear over the range of 0.60-8.92 microg (r = 0.9997), the average recovery of the method was 98.4%. RSD 1.8% (n = 6).

CONCLUSIONThe results showed that method was reliable and accurate.

Animals ; Cholesterol Esters ; analysis ; Chromatography, High Pressure Liquid ; methods ; Female ; Materia Medica ; analysis ; chemistry ; Oviducts ; chemistry ; Quality Control ; Rana temporaria

OBJECTIVEStudies on the substance with nourishment Yin for a reasonable and rational auality appraise for turtleback.

METHODTo separate by chromatography and identifying with MS, 1H NMR, 13C NMR, IR.

RESULTTwo compounds were separated and identified as hexadecanoyl cholesterol ester and cholesterol.

CONCLUSIONThe two compounds are isolated from turtleback for the first time.

Animals ; Cholesterol ; isolation & purification ; Cholesterol Esters ; chemistry ; isolation & purification ; Materia Medica ; chemistry ; Turtles

Abstract: Our previous work revealed berberine can significantly enhance the susceptibility of fluconazole against fluconazole-resistant Candida albicans, which suggested that berberine has synergistic antifungal activity with fluconazole. Preliminary SAR of berberine needs to be studied for the possibility of investigating its target and SAR, improving its drug-likeness, and exploring new scaffold. In this work, 13-substitutited benzyl berberine derivatives and N-benzyl isoquinoline analogues were synthesized and characterized by 1H NMR and MS. Their synergetic activity with fluconazole against fluconazole-resistant Candida albicans was evaluated in vitro. The 13-substitutited benzyl berberine derivatives 1a-1e exhibited comparable activity to berberine, which suggested that the introduction of functional groups to C-13 can maintain its activity. The N-benzyl isoquinolines, which were designed as analogues of berberine with its D ring opened, exhibited lower activity than berberine. However, compound 2b, 2c, and 4b showed moderate activity, which indicated that berberine may be deconstructed to new scaffold with synergistic antifungal activity with fluconazole. The results of our research may be helpful to the SAR studies on its other biological activities.
Antifungal Agents ; pharmacology ; Berberine ; pharmacology ; Candida albicans ; drug effects ; Drug Resistance, Fungal ; Drug Synergism ; Fluconazole ; pharmacology ; Isoquinolines ; pharmacology ; Microbial Sensitivity Tests

OBJECTIVETo evaluate the utility of neuronavigation in the microsurgery for cerebral cavernous malformations.

METHODSIn a retrospective study, 47 patients with cavernous malformations were involved from January 1995 to December 2010. Fourteen cases admitted into hospital from January 1995 to December 2002 were treated without neuronavigation (conventional group), the focus localization was based on magnetic resonance imaging (MRI) images, anatomic landmarks, and the experiences of the neurosurgeon. In the other 33 cases admitted after February 2002 to December 2010, surgeries were performed with neuronavigation using the Medtronic Stealth Station TREON or TRIA system (neuronavigation group). Excision of the cavernomas were all performed microsurgically, surrounding gliotic rim and hemosiderin stained tissue were resected in the case of epilepsy, and a few patients underwent extended hippocampal resection or multiple subpial transection.

RESULTSWith the use of neuronavigation, the extent of craniotomy reduced from 5.2 cm to 3.6 cm (P < 0.01), and deeper cavernoma focuses could be treated surgically. There were no changes with regard to the mean size of the cavernomas, the mean time of surgery and hospital stay (P > 0.05), but the mean time of anaesthesia was prolonged from 164 min to 197 min (P < 0.01). Cavernomas were resected completely in all 47 cases, which was confirmed by postoperative MRI recheck. The conditions of all patients were improved or remain unchanged, and no significant differences in the clinical outcome could be evaluated between the two groups.

CONCLUSIONSApplication of neuronavigation-assisted microsurgery of intracranial cavernous malformations surgery, helps reduce the surgical trauma, and has security and reliability.

Adolescent ; Adult ; Female ; Hemangioma, Cavernous, Central Nervous System ; surgery ; Humans ; Male ; Microsurgery ; methods ; Middle Aged ; Neuronavigation ; Retrospective Studies ; Young Adult

OBJECTIVETo investigate the effect of multi-plane Hyaluronic acid (HA) injection for rhinoplasty.

METHODSThe HA was injected below or above the periosteum at the nasal bone, above the perichondrium at the cartilage portion of nose, and between the great alar cartilage at the nasal tip. The HA volume was 1-1.5 ml, according to the nose form and aesthetic assessment. Over-injection was not permitted. Touch-up injection could be performed one week after the first injection if need.

RESULTSFrom Jan. 2010 to Jan. 2012, 60 cases underwent rhinoplasty with HA injection. The patients were followed up for 10-13 months with satisfactory result. The effect lasted about 9 months with the longest period as 12 months and the shortest period as 6 months.

CONCLUSIONSGood results can be achieved with multi-plane HA injection for rhinoplasty.

Adult ; Female ; Follow-Up Studies ; Humans ; Hyaluronic Acid ; administration & dosage ; therapeutic use ; Injections ; Male ; Rhinoplasty ; methods ; Treatment Outcome ; Young Adult

OBJECTIVETo determine the origin of Rana temporaria for quality Oviduetus Ranae in the light of historical documents and modern researches on the classification of Rana temporaria chensinensis.

METHODWorks of Chinese meteria medica of all ages, related historical documents and reports from home and abroad on researches of R. temporaria chensinensis were consulted, sorted out, analyzed and summarized.

RESULTThe original Shange recorded in the works of Chinese meteria medica is R. temporaria chensinensis, which is the independent species, not one of species of European forest frogs. R. temporaria chensinensis is divided into 4 subspecies: R. temporaria chensinensis, Lanzhou, Kangding, and Changbaishan. The origin of R. temporaria is Changbaishan subspecies of R. temporaria chensinensis.

CONCLUSIONChangbaishan subspecies of R. temporaria chensinensis is determined as the origin for quality Oviduetus Ranae.

Animals ; Female ; History, 18th Century ; History, 19th Century ; History, 20th Century ; History, Ancient ; Materia Medica ; history ; isolation & purification ; Oviducts ; chemistry ; Rana temporaria ; anatomy & histology ; Ranidae ; anatomy & histology ; classification ; Species Specificity ; Terminology as Topic

BACKGROUNDThere are few researches for the healing of metaphyseal fractures; moreover, the animal models to study the metaphyseal fractures are usually made by the oscillating saw osteotomy without reliable fixation, which is not in accordance with our current clinical practice. In this study, we established a new model to observe the healing process of metaphyseal fractures.

METHODSEighteen New Zealand rabbits were used in the study. The fracture model was created by splitting the medial tibial plateau in rabbits, then reset, and fixed with compression screws. At 1, 2, 3, 4, 6, and 8 weeks postoperatively, the tibial specimens were collected; firstly, a general observation and an X-ray examination of the specimens was done, and then they were embedded in methylmethacrylate and cut into sections with hard tissue slicer. The sections were stained with Giemsa reagent and examined under light microscopy.

RESULTSThere was no fracture displacement in the tibial specimens of all time points, except for one showing a collapse. No external callus formation could be observed by X-ray and general examination. After 1 week of the operation, the fracture gap was filled by mesenchymal tissue; 2 weeks postoperatively, a large number of woven bones were formed; from the third week onwards, the woven bone began to turn into lamellar bone, and new trabecular structure began to form. In all of the slices, no obvious chondrocytes formed in fracture areas; thus, there was no endochondral ossification.

CONCLUSIONSThis model was an ideal fracture animal model and suitable for the study of metaphyseal fracture healing. The X-ray and histological images demonstrated that metaphyseal fracture healing was a process of direct bone healing through intramembranous bone formation under the conditions of minor trauma, good reduction, and firm fixation.

Animals ; Fracture Healing ; physiology ; Fractures, Bone ; diagnostic imaging ; pathology ; Rabbits ; Radiography

The discovery of high performance leading antidiabetic compounds containing sulfonamide and 4-aminophenylacetic acid moieties is reported. This was achieved by the synthesis of 6 intermediates and subsequently 20 target molecules using 4-aminophenylacetic acid as the starting materials, and through a few synthetic routes aided by multi-step reactions including sulfonylation of amino group, deacylation of amides and esterification of carboxyl group, as well as acylation of amino group. The chemical structures of the twenty-four new compounds were determined using 1H NMR, 13C NMR and HR-MS techniques. Screening in vitro of their peroxisome proliferator-activated receptor (PPAR) activation activities showed weak relative PPAR activation activities to most of the target molecules. However, 4 target molecules exhibit PPAR over 58%, and as high as 81.79% for TM2-i, presenting itself as potent leading compound for antidiabetic drugs. This research also confirms that it is probable to achieve esterification of carboxyl group and deacylation of fatty acid N-phenyl amides concurrently in SOCl2/alcohol solvent system. This provides new synthetic method for the selective reaction within molecules containing both carboxyl and N-aryl amido groups of fatty acids.
Aniline Compounds ; chemistry ; Fatty Acids ; chemistry ; Hep G2 Cells ; metabolism ; Humans ; Hypoglycemic Agents ; chemical synthesis ; chemistry ; pharmacology ; Molecular Structure ; Peroxisome Proliferator-Activated Receptors ; metabolism ; Phenylacetates ; chemical synthesis ; chemistry ; pharmacology ; Structure-Activity Relationship ; Sulfonamides ; chemistry

OBJECTIVETo analyze the impact of efforts of community-based organizations (CBO) in HIV testing mobilization and case finding among men who have sex with men(MSM).

METHODSResults of HIV testing mobilization among MSM through CBOs in 15 program areas were collected and compared with corresponding HIV case reporting data to demonstrate the contribution of CBO-based HIV testing in HIV case finding among MSM from July 2008 to December 2011. Meanwhile,the proportion of screened HIV positives who received testing results notification,confirmatory test, following up and CD4 cell tests were analyzed and compared with those identified in medical institutions.

RESULTSA total of 196 075 HIV tests were performed for MSM, as a result of mobilization efforts of CBOs. Cumulatively 7704 new HIV cases were identified, accounting for 51.7% (7704/14 914) of all newly diagnosed HIV cases infected via homosexual sex in the program areas.Among the newly diagnosed MSM HIV infections in the program areas,the proportion of infections detected through the mobilization of CBOs increased from 35.4% (609/1722) in 2008 to 63.7% (2371/3722) in 2010, and 58.3% (3024/5189) in 2011. Compared with those identified through medical institutions, newly diagnosed MSM infections detected though CBOs testing mobilization have higher rates of receiving screening testing results notification (97.3% (4441/4563) vs 92.8% (13 140/14 153)) , (84.6% (2559/3024) vs 79.8% (5589/7002)) and CD4 cell tests (66.1% (1999/3024) vs 52.9% (3705/7002)), and a lower rate of receiving confirmatory test (78.6% (3588/4563) vs 85.6% (12 115/14 153)).

CONCLUSIONCBOs can take their advantages in mobilizing MSM to receive HIV test, and MSM HIV cases detected through CBOs have become the main source of MSM HIV case finding in program areas.

Community Health Services ; HIV Infections ; prevention & control ; HIV Seropositivity ; Health Promotion ; Homosexuality, Male ; Humans ; Male ; Mass Screening