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.

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.

Objective To observe the clinical efficacy and safety of alendronate tablets combined with alfacalcidolon capsules in the treatment of perimenopausal women with osteoporosis.Methods A total of 56 perimenopausal women with osteoporosis were randomly divided into control group and treatment group with 28 cases per group.Control group was treated with alfacalcidolon capsule 0.25 μg qd oral.Treatment group was given alendronate sodium tablet 70 mg qd oral,on the basis of control group.The chnical efficacy,serum aquaporin 1 (AQP1),matrix metalloproteinase 2 (MMP-2),bone alkaline phosphatase (BALP),type Ⅰ collagen C end peptide (CTX-Ⅰ)and adverse drug reactions were compared between two groups.Results After treatment,the total effective rates in treatment and control groups were 92.86％ (26/28 cases) and 71.43％ (20/28 cases) with significant difference (P ＜0.05).After treatment,the main indexes in treatment and control groups were compared:AQP1 were (23.53 ± 3.02),(30.65 ± 4.31) μg · L-1;MMP-2 were (60.37 ± 8.35),(66.45 ± 8.47) μg · L-1;BALP were (20.04±2.57),(24.38 ±3.32) pg · L-1;CTX-Ⅰ were (0.75 ±0.06),(0.83 ±0.09)ng · L-1,the differences were statistically significant (P ＜ 0.05).The adverse drug reactions in treatment group were based on diarrhea and nausea,which in control group was based on diarrhea and nausea.The incidence of adverse drug reactions in two groups were 7.14％ without significant difference (P ＞ 0.05).Conclusion Alendronate tablets combined with alfacalcidolon capsules have a definitive clinical efficacy in the treatment of perimenopausal women with osteoporosis,which can significantly reduce the serum levels of AQP1,MMP-2,BALP and CTX-Ⅰ,without increasing the incidence of adverse drug reactions.

Background:Reduced expression of tripartite motif-containing 3 (TRIM3) has been reported to be involved in the pathogenesis of human glioblastoma.In our previous research,we found that TRIM3 expression was markedly reduced in human primary hepatocellular carcinoma (HCC) tissues and that low TRIM3 expression was associated with short survival of HCC patients.However,the role of TRIM3 in liver cancer remains unknown.This study aimed to investigate the function of TRIM3 in liver cancer cells.Methods:The protein levels of TRIM3 in five liver cancer cell lines (SK-Hep1,Hep3B,Huh7,HepG2,Bel-7402) and one normal liver cell line (L02) were detected with Western blotting.HepG2 and Bel-7402 cells with IowTRIM3 expression were infected with recombinant lentiviruses overexpressing TRIM3 (LV-TRIM3),whereas Huh7 and Hep3B cells with high TRIM3 expression were transfected with TRIM3-targeted small interfering RNA (siTRIM3).The functions of TRIM3 in the proliferation,colony formation,cell cycle,migration,invasion,and apoptosis of the above cell lines were examined.The effect of TRIM3 on tumor growth and metastases in nude mice was also investigated.Results:TRIM3 was overexpressed in HepG2 and Bel-7402 cells with LV-TRIM3 infection,which further reduced proliferation,colony formation,migration,and invasion of both cell lines.Cell cycle analysis showed thatTRIM3 overexpression induced G0/G1 phase arrest in HepG2 and Bel-7402 cells.Moreover,apoptosis was not increased in HepG2 or Bel-7402 cells overexpressing TRIM3.Contrarily,silencing TRIM3 expression in Huh7 and Hep3B cells by siTRIM3 led to significantly decreased percentages of both cells in the G0/G1 phase and promoted cell proliferation,colony formation,migration,and invasion.In vivo experiment results confirmed thatTRIM3 overexpression suppressed tumor growth and metastasis.Conclusions:TRIM3 plays a tumor-suppressing role in the regulation of liver cancer development by reducing cell proliferation through cell cycle arrest at the G0/G1 phase.

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 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.