The purification of kallikrein using CTAB/hexanol/octane reverse micelles extraction has been studied and optimized,under various aqueous pH values, ionic strength and species, CTAB concentration and co-surfactant concentration. The result shows that the extraction efficiency approaches 100%, and the activity recovery is more than 80%, the commercial enzyme specific activity is increased by 1.97 times and the crude enzyme activity is increased by 7.15 times, which from 31U/mg to 219U/mg,under the conditions of[CTAB]=0.02 mol/L, hexanol/octane (V/V)=1∶5, pH=9.0 and[KBr]=0.1 mol/L in forward extraction, pH=7.0 ,[KBr]=1.5 mol/L and 15% ethanol(V/V) in backward extraction. The result of purified kallikrein is examined by the SDS-PAGE analysis. Reverse micelles extraction is a potential technique for the application in the downstream biotechnological processes.

Guanylate-binding protein 1 (GBP1) is an interferon induced protein, that belongs to the guany late-binding protein family. GBP1 is widely involved in anti-infection immune responses, anti-tumor activity and various biological reactions. Recent studies have proved that IFN-alpha, IFN-beta, IFN-gamma, IL1alpha, IL1beta, TNF-alpha and LPS can induce GBP1 expression; hence, the diverse biological functions of GBP1 have been gradually deduced and exploited. Many studies have been performed over recent years to understand the exact mechanisms that underlie the anti-infection and anti-tumor properties of GBP1. This review describes the molecular structure, biological activity, anti-infective properties and other functions of GBP1, in order to provide insights into the divergent roles of GBP1 in the regulation of various biological processes.
Animals ; Antineoplastic Agents ; metabolism ; Antiviral Agents ; metabolism ; GTP-Binding Proteins ; chemistry ; genetics ; metabolism ; Humans ; Interferons ; genetics ; metabolism

Bacillus subtilis B6-1 was used as an original strain for mutagenic treatment and a defined medium was used as the selective medium. A mutant named B. subtilis W003 was isolated after three serial ultraviolet (UV) irradiations and one diethyl sulfate (DES) treatment. The ?-PGA yield on a rotary shaker was enhanced from 10.9 g/L in parental strain to 20.5 g/L in the mutant. It was illustrated by single factor experiments that the optimal carbon and nitrogen sources were glucose and (NH4)2SO4 respectively. The optimal fermentation medium was achieved by orthogonal test. In the optimal medium, a ?-PGA yield of 45.3 g/L was obtained after 36 h cultivation.

OBJECTIVE Cistanche deserticola Y. C. Ma is an official plant that grows in arid or semi-arid areas.Our early work demonstrated that Cistanche extracts protect against sperm damage in mice under bisphenol A induced reproductive damage. Echinacoside (ECH)is one of the major active components. The present study investigated the mechanisms behind the possible protective effects of ECH against oligoasthenospermia in rat and identified the interaction between ECH and AR. METH-ODS The distribution of ECH was assayed by HPLC,the quantity and quality of sperm was evaluated and hormone levels were determined by radioimmunosorbent assay. The levels of androgen receptor (AR)and key steroidogenic-related genes were reduced as determined by Western blotting and qPCR analysis.The interaction between ECH and AR were evaluated by fluorescence localization assay,indi-rect ELISA and molecular docking. RESULTS ECH significantly increased the quantity of sperm and secretions of luteinizing hormone and testosterone.ECH was distributed to the hypothalamus but not in the testes.ECH combined with hypothalamic AR in the pocket of Met-894 and Val-713 to inhibit transfer of AR from the cytoplasm to nuclei in the hypothalamus.While negative feedback of sex hormone regula-tion was inhibited, positive feedback was stimulated to increase the secretion of luteinizing hormone and testosterone subsequently enhancing the quantity of sperm. C. militaris significantly alleviated the BPA-induced reproductive damage by increasing testicular superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and glutathione (GSH); as well as by reducing serum malondialdehyde (MDA). C. militaris not only obviously enhanced the levels of serum LH and T, but it also improved the sperm count and motility compared to the BPA-treated group.CONCLUSION C.militaris and ECH protect the BPA induced reproductive damage.ECH blocks AR activity in the hypothalamus to increase the quantity of sperm and protect against oligoasthenospermia in rats.

Worldwide sales of biologic drugs exceeded 100 billion USD in 2011. About 32% is from therapeutic monoclonal antibody (mAb). With many blockbuster biopharmaceutical patents expiring over the next decade, there is a great opportunity for biosimilar to enter the worldwide especially emerging market. Both European Medicines Agency (EMA) and Food and Drug Administration (FDA) have introduced regulatory frameworks for the potential approval of biosimilar mAb therapeutics. Rather than providing a highly abbreviated path, as in the case for small molecule chemical drug, approval for biosimilar mAb will require clinical trial and the details will be very much on a case-by-case basis. Since mAb is the dominant category of biologic drugs, mAb will be the focus of this review. First, the United States (US) and European Union (EU) approved mAb and those in phase 3 trials will be reviewed, then strategies on how to win biosimilar competition will be reviewed.
Animals ; Antibodies, Bispecific ; therapeutic use ; Antibodies, Monoclonal ; therapeutic use ; Antibodies, Monoclonal, Humanized ; therapeutic use ; Biosimilar Pharmaceuticals ; standards ; Clinical Trials, Phase III as Topic ; Drug Approval ; European Union ; Humans ; United States ; United States Food and Drug Administration

Adult ; Aged ; Bone Nails ; Female ; Femoral Fractures ; surgery ; Femur ; surgery ; Fracture Fixation, Intramedullary ; Fractures, Comminuted ; surgery ; Humans ; Internal Fixators ; Male ; Middle Aged

OBJECTIVETo investigate the effects of prostacyclin (PGI(2)) and nitric oxide (NO) in the development of hyperdynamic circulatory state on chronic portal hypertensive rats.

METHODSSixty-six male SD rats were divided into three groups, namely intrahepatic portal hypertension (IHPH) by injection of CCl(4), prehepatic portal hypertension (PHPH) by partial stenosis of the portal vein for 2 weeks and sham-operated controls (SO). Animals in each group were divided further into 3 subgroups and received N(omega)-nitro-L-arginine (L-NNA), indomethacin and saline (as control), respectively. Splanchnic and systemic hemodynamics was measured using radioactive microsphere techniques. The NO concentration in serum was determined by nitrates-nitrites which were measured using a colorimetric method, and concentration of PGI(2) was determined using specific radioimmunoassay for its stable hydrolytic product, 6-keto-PGF(1 alpha).

RESULTSThe concentrations of plasma 6-keto-PGF(1 alpha) and serum nitrates + nitrites in IHPH rats (1 123.85 +/- 153.64; 73.34 +/- 4.31) and in PHPH rats (891.88 +/- 83.11; 75.21 +/- 6.89) were significantly higher than those of SO rats (725.53 +/- 105.54;58.79 +/- 8.47). L-NNA and indomethacin could decrease the concentrations of plasma 6-keto-PGF(1 alpha) and serum nitrates + nitrites in IHPH and PHPH rats (P < 0.05). At the same time, CI, FPP and PVI were lowered while MAP, TPR and SVR were increased (P < 0.05). After deduction of NO action, there were no significant correlation between plasma PGI(2) level and hemodynamic parameters such as CI, TPR, PVI and SVR. However, after deduction of PGI(2) action, NO was still correlated highly with those hemodynamic parameters.

CONCLUSIONIt is NO rather then PGI(2) that is a mediator in the formation and development of hyperdynamic circulatory state in chronic portal hypertensive rats.

6-Ketoprostaglandin F1 alpha ; blood ; Animals ; Cyclooxygenase Inhibitors ; pharmacology ; Disease Models, Animal ; Enzyme Inhibitors ; pharmacology ; Epoprostenol ; blood ; physiology ; Hemodynamics ; drug effects ; Hypertension, Portal ; blood ; physiopathology ; Male ; Nitric Oxide ; blood ; physiology ; Nitric Oxide Synthase ; antagonists & inhibitors ; Nitroarginine ; blood ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; omega-N-Methylarginine ; pharmacology

OBJECTIVETo observe the effect of single herb pilose antler (PA) on the expression of Smad2 and Smad3 in the cartilage of osteoarthritis (OA) rats.

METHODSOne hundred 3-month old female healthy SD rats, (200 +/- 20) g, were recruited and routinely fed for 1 week. They were randomly divided into 5 groups, i.e., the low dose PA group, the high dose PA group, the normal saline control group, the model group, and the normal control group, 20 in each group. The model was prepared using classic Hulth method except the normal control group. After 6-week modeling, the model was confirmed successful by pathologic observation. PA at 0.021 g/100 g and 0.084 g/1 00 g was given by gastrogavage to rats in the low dose PA group and the high dose PA group respectively. Normal saline was administered to those in the normal saline control group. No treatment was given to rats in the normal control group and the model group. Bilateral knee cartilages were harvested at week 2,4, and 6. mRNA and protein expressions of Smad2 and Smad3 were detected by immunohistochemical assay, fluorescent quantitative PCR, and Western blot.

RESULTSOA model was successfully prepared by pathological observation. Results of immunohistochemical assay showed that Smad2 and Smad3 expressed extensively in the cartilage, and located inside the chondrocyte membrane. Compared with the model group, mRNA expression of Smad2 and Smad3 obviously increased in the low dose PA group and the high dose PA group at week 2, 4, and 6, showing statistical difference (P < 0.05). Compared with the same group at week 4 after gastrogavage, mRNA expression of Smad2 and Smad3 obviously decreased in the low dose PA group and the high dose PA group at week 6, showing statistical difference (P < 0.05). Compared with the model group, protein expression of Smad2 and Smad3 obviously increased in the chondrocytes of the low dose PA group and the high dose PA group at week 2 and 4, showing statistical difference (P < 0.01). Compared with the same group at week 2 after gastrogavage, protein expression of Smad2 and Smad3 obviously increased in the low dose PA group and the high dose PA group at week 4, showing statistical difference (P < 0.01). Compared with the same group at week 4 after gastrogavage, protein expression of Smad2 and Smad3 obviously decreased in the low dose PA group and the high dose PA group at week 6, showing statistical difference (P < 0.01).

CONCLUSIONS(1) The pilose antler could repair cartilages by regulating mRNA and protein expressions of Smad2 and Smad3. (2) Up-regulating mRNA and protein expressions of Smad2 and Smad3 might be one of important mechanisms for the pathogenesis of OA.

Animals ; Antlers ; chemistry ; Cartilage ; cytology ; metabolism ; Chondrocytes ; drug effects ; metabolism ; Female ; Medicine, Chinese Traditional ; Osteoarthritis ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley ; Smad2 Protein ; metabolism ; Smad3 Protein ; metabolism

OBJECTIVETo investigate the computed tomography (CT) manifestations of liver involvement in children with Langerhans cell histiocytosis (LCH).

METHODSRetrospective analysis was performed on 9 LCH children with liver involvement confirmed by clinical, laboratory and pathological examinations to investigate the CT manifestations of this condition. These children, including 6 males and 3 females, had undergone both plain CT scan and dual-phase (the arterial and portal venous phases) contrast-enhanced CT scan.

RESULTSThe main CT manifestations included hepatomegaly (8 cases); periportal dendritic hypodense lesions or "periportal halo sign" (7 cases) which were mildly or moderately enhanced in the arterial phase; intrahepatic bile duct dilatation (5 cases); lymphadenopathy in the hepatic hilar or retroperitoneal region (4 cases); and diffuse small hypodense nodules (3 cases), which showed annular enhancement on the contrast-enhanced CT scan.

CONCLUSIONSCT findings may be helpful in the early diagnosis and treatment of LCH in children.

Child ; Child, Preschool ; Female ; Hepatomegaly ; diagnostic imaging ; Histiocytosis, Langerhans-Cell ; complications ; diagnostic imaging ; Humans ; Infant ; Liver ; diagnostic imaging ; pathology ; Male ; Retrospective Studies ; Tomography, X-Ray Computed ; methods

Adult ; Female ; Humans ; Male ; Medial Collateral Ligament, Knee ; injuries ; surgery ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Rupture ; Suture Anchors