Adiponectin is an adipocyte-derived secretory protein. It was found to be associated with insulin resistance, inflammation and arteriosclerosis. To further study the biological function and expression of adiponection in vivo, adipoenctin gene knock-out and LacZ gene knock-in mouse model was constructed. Gene targeting strategy was designed to replace part of exon 2 and exon 3 of adiponectin gene with full length LacZ gene in frame with remaining upstream ATG and signal peptide sequence of exon 2. The targeting vector (Adipo-LacZ-XpPNT) was constructed and verified by restriction enzyme digestion and sequencing. CJ7 ES cells were transfected with targeting vector linearized by NotⅠ digestion, selected in the medium containing both G418 and ganciclovoir. Resistant clones were screened by PCR and further confirmed by Southern blot for correct homologous recombinants. Chimera mice were obtained by routing microinjection of homologous recombined ES cells into blastocysts. After mating, mice heterozygous and further homozygous for adiponectin knockout and LacZ gene knock-in were established. Expression of both endogenous adiponectin and exogenous LacZ gene in mouse tissues and sera were detected by RT-PCR, Northern-blot, Western blot and ELISA. The results show that adiponectin was disrupted at both mRNA and protein levels. LacZ gene is expressed exclusively in adipose tissue of mutant mice. Its expression profile is identical to endogenous adiponection. Unexpectedly, LacZ activity could not be detected in both adipose tissue and serum although LacZ protein can be detected in adipose tissue but not in serum of mutant mice. In conclusion, mice homozygous for adiponectin knockout and LacZ gene knock-in have been successfully constructed. Mutant mice display LacZ expression profile identical to endogenous adiponectin albeit neither LacZ activity nor protein can be detected in serum of mutant mice.

Objective To study the clinical and imaging features of patients with bone metastases from breast cancer and identify the factors related to the incidence of bone metastases. Methods Three hundred and thirty-four patients with breast cancer were recruited into this study. Whole-body 99Tcm-methylene disphosphonate (MDP) bone scan, clinical staging, pathological, immunohistochemical and serological test results were analyzed retrospectively. χ2 test was used for statistical analysis. Results The incidence rate of bone metastases for patients with and without lymph node metastases was 71% (152/214) and 22. 5% (27/120), respectively (χ2 =72.80, P =0.000). The incidence rate of bone metastases from infiltrated non-specified and specified breast cancer was 69% (203/294) and 41.7% (5/12), respectively (χ2 =3. 97, P=0.046). Alkaline phosphatase (ALP) was elevated in 28.5% (51/179) and 14.9%(11/74) of patients with and without bone metastases, respectively (χ2 = 5. 25, P = 0.022 ). Carcinoembryonic antigen (CEA), carbohydrate antigen (CA) 15-3, CA125, CA19-9 increased in 68.7% ( 123/179) and 27.0% (20/74) of patients with and without bone metastases, respectively (χ2 = 37. 03, P =0. 000). Conclusions The incidence of bone metastases from breast cancer is correlated to pathological types of primary tumor and lymph node metastases. Bone metastases occurs more frequently in patients with infiltrated, non-specified, primary cancer and with lymph node metastases. Serum ALP, CEA, CA15-3,CA125, CA19-9 might be the tumor makers for early diagnosis of bone metastases from breast cancer.

OBJECTIVETo observe the protective effects of the Huanglian Jiedu decoction aqueous extract and its active fraction, which consists of total alkaloids, total flavonoids and total iridoid, on the thalamus of cerebral ischemia in rats.

METHODThe rat model of middle cerebral artery occlusion (MCAO) was chosen. Male SD rats were randomly divided into sham-operation group, model group, aqueous extract group (800 mg x kg(-1)), total alkaloids group(44 mg x kg(-1)), total flavonoids group (50 mg x kg(-1)) and the total iridoid group (80 mg x kg(-1)). The rats were administered the appropriate drugs intragastrically once a day, for 7 days after surgery. An equivalent volume of saline was given in the sham surgery and model groups. The HE staining was adopted to observe the pathological changes. Determination of Glu and gamma-GABA in thalamus were detected by HPLC with fluorescence detection. The expression of GAD65 was examined with immunohistochemistry and double staining with uorescent-conjugated antibodies against GFAP and Cx43 was chosen in this study.

RESULTThe neurons degenerated in MCAO rats after cerebral ischemia 7 d. The content of Glu, gamma-GABA decreased (P < 0.05), the expression of GAD65 reduced (P < 0.05) and the expression of GFAP and Cx43 increased (P < 0.01) in thalamus of rats compared with sham-operation group. Huanglian Jiedu decoction aqueous extract, total alkaloids, total flavonoids and total iridoid reduced the degeneration of neurons. Total flavonoids could promote the expression of GAD65 (P < 0.05) and decrease the expression of GFAP and Cx43 (P < 0.01) in thalamus compared with model group while it could also increased the content of Glu,gamma-GA BA to normal levels. Compared with model group, Huanglian Jiedu decoction aqueous extract, total alkaloids and total iridoid could raise the expression of Cx43, and Huanglian Jiedu decoction aqueous extract could also increase the expression of GAD65 (P < 0.05). The expression of GFAP in Huanglian Jiedu decoction aqueous extract group, total alkaloids group and total iridoid group were not different compared with model group while the content of gamma-GABA decreased (P < 0.05) compared with sham-operation group.

CONCLUSIONThe degeneration of nerve cells, the reduction of neurotransmitter amino acids content, the aberrant activation of astrocytes and the abnormal expression of GFAP and Cx43 will appear in thalamus of MCAO rats after ischemia. Huanglian Jiedu decoction total flavonoids could relieve the injury of nerve cell through inhibiting the abnormal activation of astrocytes and regulating the expression of GFAP and GAD65.

Animals ; Astrocytes ; drug effects ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Infarction, Middle Cerebral Artery ; drug therapy ; Male ; Protective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Thalamus ; drug effects

Objective To understand the pathogenic distribution and epidemiological trend of hand-foot-and-mouth disease (HFMD),and provide evidence for the prevention and control of HFMD.Methods Children who were diagnosed with HFMD in a hospital between January and December 2015 were investigated,real time fluorescence PCR was used to detect enterovirus universal type EV,enterovirus 71 (EV71),and Coxsackievirus A16 (CoxA16) in specimens from children with HFMD.Positive rates and distribution of various types of EV among children of different months,genders,age groups,and infection types were analyzed.Results A total of 837 throat swab specimens from HFMD children were collected in 2015,380 (45.40％) of which were EV positive specimens.Virus typing showed that 110 (28.95 ％),7 (1.84 ％),6(1.58 ％),and 257(67.63 ％) were positive specimens for EV71,CoxA16,EV71 + CoxA16,and other types of EV.HFMD had a high prevalence since April,reached a peak in May-June,and remained high incidence in July-December.Positive rates of EV in children of different months were statistically different (P＜0.05).The age of onset was mainly in children under 3 years.Positive rates of EV and constitute ratios of different types of EV in children of different age groups were all statistically different (all P＜0.05).The positive rate of EV in severe HFMD cases was higher than common cases (65.34％ vs 27.06％,P＜0.001).The proportion of severe cases in children with EV71 infection and other types of EV infection were 90.00％ and 60.70％ respectively;children with EV71 + CoxA16 double infection were all severe cases.Constitute of EV types in children with different infection types was statistically different(P＜0.001).Conclusion In 2015,EV infection in hospitalized children with HFMD in this hospital was mainly caused by other types of EV (nonEV71 and non-CoxA16),the high prevalence season,high-risk population under 3 years of age,and severe cases should be paid high attention,prevention and treatment should be performed well.

OBJECTIVETo prepare sustained-release microsphere containing extract of Sanguis Draconis and to measure its dissolution in vitro.

METHODSustained-release microsphere was prepared with polylactic acid (PLA) as carriers using the oil-in-water (O/W) emulsion solvent evaporation method. The powder particle's characteristics of sustainded-release microsphere were evaluated comprehensively, and its dissolution characteristics in vitro were studied.

RESULTThe microsphere was round and its surface was smooth, drug-loading rate was 21.97% and the entrapment rate was 55.76%, the accumulative release percentage was 76. 71% in 16 hours.

CONCLUSIONThe sustained release effect of Sanguis Draconis microspheres was formed with potentially wide applications.

Arecaceae ; chemistry ; Delayed-Action Preparations ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Lactic Acid ; chemistry ; Microspheres ; Particle Size ; Plants, Medicinal ; chemistry ; Polyesters ; Polymers ; chemistry ; Reproducibility of Results ; Resins, Plant ; chemistry ; Technology, Pharmaceutical ; methods

OBJECTIVETo explore a new method and estimate its outcome to repair deep heel tissue injuries with local tissue flap.

METHODSAt lower cnemis, a fat fascial flap combined with perineal tendofascial flap were designed and elevated between the lateral malleolars blank, the largest area is 6 cm x 13 cm, which were transplanted to cover naked calcaneal tendon and calcaneal bone, full-thickness free skin graft or middle-thickness free skin graft were grafted on flap.

RESULTS6 patients suffered from heel tissue defects had been treated from January 2004 to November 2005. The smallest area was 3.5 cm x 4.0 cm and the largest was 4 cm x 6 cm. All fascial flaps and free skin grafts were successfully survived after operations. All patients were followed up 3-23 months, the operation areas were healed, the functions and configurations were satisfied.

CONCLUSIONSPerineal tendofascial flap conjoined with adipofasical flap and free skin graft are used to repair heel deep tissue defects, which is a handy, dependable way for repairing of the small tissue defects of the lower third of leg and heel.

Achilles Tendon ; injuries ; surgery ; Adult ; Fascia ; transplantation ; Female ; Fibula ; surgery ; Heel ; injuries ; surgery ; Humans ; Male ; Middle Aged ; Skin Transplantation ; Soft Tissue Injuries ; surgery ; Surgical Flaps

The aim of this study was to explore the difference of MSC migration mediated by SDF-1/CXCR4 axis through Boyden chamber in vitro migration assay. The SDF-1 density-dependence of MSC migration was observed. Subsequently, the effects of different blocking agents on hSDF-MSC migration were observed after MSC were treated with 50 nmol/L wortmannin, 10 micromol/L LY294002, 50 micromol/L PD98059, 10 micromol/L U73122, 126 micromol/L AMD3100 and 50 nmol/L verapamil respectively. The results showed the efficiency of MSC migration increased gradually with the increasing of hSDF-1 density. And after MSCs treatment with 50 nmol/L wortmannin, 10 micromol/L LY294002, 50 micromol/L PD98059, 10 micromol/L U73122 and 126 micromol/L AMD3100 respectively, the ability of MSC migration decreased. The ability of MSCs migration obviously decreased when MSCs were treated with U73122, AMD3100. It is concluded that the SDF-1/CXCR4-mediated MSC migration may be related to mitogen-activated protein kinase (MAPK), phosphatidylinositol phospholipase C (PI-PLC) and protein kinase (PKC) signal pathways.
Animals ; Bone Marrow Cells ; cytology ; Cell Movement ; Flavonoids ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; Protein Kinase C ; metabolism ; Rats ; Rats, Wistar ; Receptors, CXCR4 ; metabolism ; Signal Transduction

Objective To validate the analysis capability of RapidHITTM 200 system for four kinds of routine forensic samples and the recyclable capability of template, template DNA and PCR products in the process of twice duplicate detection. Methods The buccal swabs underwent the test twice by RapidHITTM 200 system, and the template DNA and PCR products that arose in the system were also tested for two times. After four kinds of routine forensic samples were detected by RapidHITTM 200 system, the follow-up tests of the template, template DNA and PCR products that arose in the system were performed. Re-sults The STR loci could be detected in the buccal swabs by the system for the first time. However, part of the STR loci lost during the second test. And the peak value obtained in the second test was significantly reduced than the one in the first time. The average STR loci detection rates of the template DNA and PCR products were both less than 50％ in the second test, which were significantly reduced than that in the first test. In addition, the analysis capability of the system for the tissues and buccal swabs was better than that for the blood and cigarette butts. Compared with the first test, the STR loci detection rate of the tested items, template DNA and PCR products decreased with the numbers of tests. Conclusion RapidHITTM 200 system is more effective in retesting buccal swabs than other samples, whereas the items, DNA template, PCR products obtained in the first and second time cannot be directly used for the further application and study of forensic medicine.

Paris is a commonly used traditional Chinese medicine (TCM), and has antitumor, antibacterial, sedative, analgesic and hemostatic effects. It has been used as an ingredient of 81 Chinese patent medicines, with a wide application and large market demand. Based on the data retrieved from state Intellectual Property Office patent database, a comprehensive analysis was made on Paris patents, so as to explore the current features of Paris patents in the aspects of domestic patent output, development trend, technology field distribution, time dimension, technology growth rate and patent applicant, and reveal the development trend of China's Paris industry. In addition, based on the current Paris resource application and development, a sustainable, multi-channel and multi-level industrial development approach was built. According to the results, studies of Paris in China are at the rapid development period, with a good development trend. However, because wild Paris resources tend to be exhausted, the studies for artificial cultivation technology should be strengthened to promote the industrial development.
Drug Development ; trends ; Drugs, Chinese Herbal ; pharmacology ; Medicine, Chinese Traditional ; Melanthiaceae ; chemistry ; Patents as Topic ; Plants, Medicinal ; chemistry

Objective To collect the kinematics and kinetics functional parameters of healthy human lower limbs during stair ascent and descent, so as to provide data for designing the gait trajectory and selecting the driving components of the exoskeleton walker to achieve the motion of stair climbing. Methods Common staircase as an experimental setup was designed, and the three-dimensional (3D) motion capture system and 3D force plate were used to simultaneously measure the ankle, knee, hip motion information during stair ascent and descent. The movement characteristics of the lower limb during stair climbing was analyzed by SPSS statistical software and Origin graphics software, and the functional parameters during stair ascent and decent were also compared and analyzed. Results The joint angle at the sagittal plane, joint moment variation with gait and activity limits in lower limbs of healthy youth during stair ascent and descent were obtained by the experiment. The ranges of motion in ankle，knee and hip joints during stair ascent were -10.66°-13.26°, 6.85°-88.92°, 1.31°-50.18°,while those during stair descent were -37.42°-27.18°,9.83°-95.53°,8.01°-31.62°.The maximum ankle, knee and hip joint moment during stair ascent and descent were 1.788, 1.121, 0.946 N•m/kg, respectively. Conclusions The gait parameters variation with gait and the joint range of motion in human lower limb were significantly different during stair ascent and descent. Separate design targeting at stair ascent or stair descent is required for gait planning of walking aids, and the selection of driving element should consider the maximum moment in each joint during stair ascent and descent.