Obej ctive Knockout mice are widely used in the studies of joint diseases .This article investigated the effects of joint processing methods , collagenase types ,and collagenase digestion time on the number of primary fibroblast -like synoviocytes (FLSs) obtained from mice. Methods The hind legs of 6 of the 12 male mice were cut open from the hip joints , but not those of the other 6.FLSs were isolated using the type-Ⅳcollagenase digestion method and purified by differential digestion .Cell morphology was observed under the inverted microscope .The type, viability, and purity of the cells were determined by flow cytometry . Rse ults Significantly fewer FLSs were obtained from the mice with the hind legs cut open ( 19 133 ±115 ) than from those without (24 933 ± 503) (P<0.05).The numbers of FLSs collected from the cell suspension at 1, 2, 3, 4, 5,6 , and 7 hours after digestion were 700 ±300 , 600 ±100 , 15 200 ±900 , 5100 ±800 ,2700 ±300 , 900 ±200, and 300 ±100, respectively, the highest at 3 hours. There were statistically significant differences in the total number of FLSs obtained by type-Ⅳ and type-Ⅱ collagenase digestions (24900 ±500v s 18 100 ±400, P<0.05). Conclusion For in virt o culture of primary mouse FLSs, it is recommended that the hip joints be not cut open, and type-Ⅳcollagenase be used with cell sus-pension at 2-6hours after digestion .

BACKGROUND/AIMS: This study investigated the protection provided by gabexate mesylate thermo-sensitive in-situ gel (GMTI) against grade III pancreatic trauma in rats. METHODS: A grade III pancreatic trauma model with main pancreatic duct dividing was established, and the pancreas anatomical diagram, ascites, and serum biochemical indices, including amylase, lipase, C-reactive protein (CRP), interleukin 6 (IL-6), and tumor necrosis factor-α (TNF-α), were examined. The pancreas was sliced and stained with hematoxylin eosin and subjected to terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. RESULTS: Ascites, serum amylase, lipase, CRP, IL-6, and TNF-α levels were significantly increased in the pancreas trauma (PT) groups with prolonged trauma time and were significantly decreased after GMTI treatment. The morphological structure of the pancreas was loose, the acinus was significantly damaged, the nuclei were irregular and hyperchromatic, and there was inflammatory cell invasion in the PT group compared to the control. After GMTI treatment, the morphological structure of the pancreas was restored, and the damaged acinus and inflammatory cell invasion were decreased compared to the PT group. Moreover, the cell apoptosis index was significantly increased in the PT group and restored to the same levels as the control group after GMTI treatment. CONCLUSIONS: GMTI, a novel formulation and drug delivery method, exhibited specific effective protection against PT with acute pancreatitis therapy and has potential value as a minimally invasive adjuvant therapy for PT with acute pancreatitis.
Amylases ; Animals ; Apoptosis ; Ascites ; C-Reactive Protein ; DNA Nucleotidylexotransferase ; Eosine Yellowish-(YS) ; Gabexate* ; Hematoxylin ; Interleukin-6 ; Lipase ; Methods ; Necrosis ; Pancreas ; Pancreatic Ducts ; Pancreatitis ; Rats*

China ; epidemiology ; Circoviridae ; genetics ; isolation & purification ; Circoviridae Infections ; epidemiology ; virology ; DNA Virus Infections ; epidemiology ; transmission ; virology ; DNA Viruses ; genetics ; isolation & purification ; DNA, Viral ; blood ; Hepatitis Viruses ; genetics ; Humans ; Nucleic Acid Probes ; Polymerase Chain Reaction ; methods ; Sequence Homology, Nucleic Acid ; Transfusion Reaction

The safety and efficacy of combined low dose aspirin and warfarin therapy in patients with atrial fibrillation after mechanical heart valve replacement were evaluated. A total of 1016 patients (620 females, mean age of 36.8±7.7 years) admitted for cardiac valve replacement and complicated with atrial fibrillation after surgery were randomly divided into study (warfarin plus 75-100 mg aspirin) or control (warfarin only) groups. International normalized ratio (INR) and prothrombin time were maintained at 1.8-2.5 and 1.5-2.0 times the normal values, respectively. Thromboembolic events and major bleedings were registered during the follow-up period. Patients were followed up for 24±9 months. The average dose of warfarin in the study and control groups was 2.91±0.83 mg and 2.88±0.76 mg, respectively (P>0.05). The incidence of overall thromboembolic events in study group was lower than that in control group (2.16% vs. 4.35%, P=0.049). No statistically significant differences were found in hemorrhage events (3.53% vs. 3.95%, P=0.722) or mortality (0.20% vs. 0.40%, P=0.559) between the two groups. Combined low dose aspirin and warfarin therapy in the patients with atrial fibrillation following mechanical heart valve replacement significantly decreased thromboembolic events as compared with warfarin therapy alone. This combined treatment was not associated with an increase in the risk of major bleeding or mortality.
Adult ; Anticoagulants ; administration & dosage ; Aspirin ; administration & dosage ; Atrial Fibrillation ; blood ; drug therapy ; etiology ; Female ; Fibrinolytic Agents ; administration & dosage ; Heart Valve Prosthesis ; Heart Valve Prosthesis Implantation ; adverse effects ; Humans ; International Normalized Ratio ; Male ; Postoperative Complications ; blood ; drug therapy ; Prothrombin Time ; Warfarin ; administration & dosage

There is evidence that 5-7 d after acute seizure episodes induced by kainic acid (KA) the rats develop a long-lasting increase in the susceptibility to seizures followed by spontaneous recurrent seizures (SRS). The present study was focused on the role of hippocampal mu opioid receptors (MORs) in the susceptibility of rats to seizures with the KA model of epilepsy. The rats received a convulsant dose of KA (10 mg/kg, i.p.) were continuously infused with a selective MOR agonist PL017 (2.09, 2.59, 3.29 microg/microl), or a selective MOR antagonist beta-funaltrexamine hydrochloride (beta-FNA, 0.88, 1.10, and 1.35 microg/microl) into ventral hippocampus by means of mini-osmotic pumps. Seven days later, the susceptibility of rats to seizures was checked by a subconvulsant dose of KA (5 mg/kg, i.p.). PL017 infusion shortened the latency and increased the stage of seizures induced by subconvulsant dose of KA in a dose-dependent manner. In contrast, infusion of beta-FNA exhibited a dose-dependent effect against seizures challenged by subconvulsant dose of KA. These results indicate that hippocampal MOR may exert a promoting effect on the susceptibility of rats to KA-induced seizures.
Animals ; Disease Susceptibility ; Dynorphins ; pharmacology ; Epilepsy ; chemically induced ; physiopathology ; Hippocampus ; physiopathology ; Kainic Acid ; Male ; Naltrexone ; analogs & derivatives ; pharmacology ; Peptide Fragments ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, mu ; agonists ; antagonists & inhibitors ; physiology

OBJECTIVETo observe the change of the neuropeptide pro-protein processing system in the ischemic retina ganglion cell-5 (RGC-5) cells, pro-protein convertase-2 (PC2), carboxypeptidase-E (CPE) and preproneuropeptide Y (preproNPY) protein levels in the ischemic RGC-5 cells and conditioned medium were analyzed.

METHODSThe RGC-5 cell was differentiated in 0.1 mumol/L staurosporine for 24 h and then stressed by different doses of oxygen and glucose deprivation (OGD). The acute or chronic OGD-induced cell death rates were obtained by using PI or TUNEL staining. The protein expression levels were determined by using the Western blot method and PC2 activity analysis.

RESULTSThe ischemia caused substantial cell death in an OGD dose-dependent manner. In the cells, proPC2 and preproNPY protein levels gradually increased whereas proCPE gradually decreased. After OGD, PC2 activity was decreased. In the conditioned medium, proPC2 and PC2 proteins gradually decreased whereas proCPE, CPE, and preproNPY proteins gradually increased.

CONCLUSIONThese results demonstrated that OGD inhibited the neuropeptide pro-protein processing system by reducing PC2 activity and the maturation of proPC2. The aggregation of the pro-proteins and the increase of the active CPE excision adversely exacerbated the cell injury. The pro-protein processing system might play a critical role in the ischemic stress of RGC-5 cells.

Animals ; Carboxypeptidase H ; metabolism ; Cell Death ; drug effects ; physiology ; Cell Differentiation ; drug effects ; Cell Hypoxia ; drug effects ; physiology ; Cell Line, Transformed ; Enzyme Inhibitors ; pharmacology ; Gene Expression Regulation, Enzymologic ; drug effects ; physiology ; Glucose ; deficiency ; In Situ Nick-End Labeling ; methods ; Indoles ; Neuropeptide Y ; metabolism ; Proprotein Convertase 2 ; metabolism ; Protein Precursors ; metabolism ; Rats ; Retinal Ganglion Cells ; drug effects ; metabolism ; Staurosporine ; pharmacology ; Time Factors

This paper summarized the Chinese literatures in the previous 5 years about the pre-clinical animal researches on the application of electro-acupuncture (EA) treatment for depression, searched in China National Knowledge Infrastructure (CNKI). The efficiency of EA treatment for depression and the mechanism of it were discussed, to shed light on new ideas and new fronts for the further research on depression in clinical or pre-clinical fields.

Extra-hypothalamic growth hormone-releasing hormone (GHRH) plays an important role in reproduction. To study the treatment effect of Grin (a novel hGHRH homodimer), the infertility models of 85 male Chinese hamsters were established by intraperitoneally injecting 20 mg/kg of cyclophosphamide once in a week for 5 weeks and the treatment with Grin or human menopausal gonadotropin (hMG) as positive control was evaluated by performing a 3-week mating experiment. 2–8 mg/kg of Grin and 200 U/kg of hMG showed similar effect and different pathological characteristics. Compared to the single cyclophosphamide group (0%), the pregnancy rates (H-, M-, L-Grin 26.7, 30.8, 31.3%, and hMG 31.3%) showed significant difference, but there was no difference between the hMG and Grin groups. The single cyclophosphamide group presented loose tubules with pathologic vacuoles and significant TUNEL positive cells. Grin induced less weight of body or testis, compactly aligned tubules with little intra-lumens, whereas hMG caused more weight of body or testis, enlarging tubules with annular clearance. Grin presented a dose-dependent manner or cell differentiation-dependentincrease in testicular GHRH receptor, and did not impact the levels of blood and testicular GH, testosterone. Grin promotes fertility by proliferating and differentiating primitive cells through up-regulating testicular GHRH receptor without triggering GH secretion, which might solve the etiology of oligoasthenozoospermia.
Animals ; Cricetinae* ; Cricetulus ; Cyclophosphamide ; Fertility ; Gonadotropins ; Growth Hormone-Releasing Hormone ; Humans ; In Situ Nick-End Labeling ; Infertility ; Infertility, Male* ; Male ; Male* ; Pregnancy Rate ; Reproduction ; Testis ; Testosterone ; Vacuoles

The authors note that on page 637 (Author Name), author affiliation of Tao Liu “Tao Liu⁶” should instead appear as “Tao Liu⁵.”

PURPOSE: The purpose of this study was to subdivide M1 stage nasopharyngeal carcinoma (NPC) patients with bone-only metastases for prognosis prediction while identifying the treatment effect of locoregional radiotherapy (LRRT) and metastasis radiotherapy (MRT) among patients with different risk. MATERIALS AND METHODS: From November 2006 to October 2016, a total of 226 patients with bone-only metastasic NPC were retrospectively enrolled. All patients developed distant lesions before receiving treatment. All potential prognostic factors were considered and the correlation of the M1 subdivisions with overall survival (OS) was determined by Cox regression hazards model. Kaplan–Meier curves were used to appraise survival condition and log-rank testing was used to compare the differences. RESULTS: The median follow-up time was 33.9 months (range, 3 to 126 months). According to multivariate Cox proportional hazard analysis, the number of metastatic lesions and Epstein-Barr virus (EBV) DNA status after palliative chemotherapy (PCT) were independent prognostic factors for OS. Thus, we subdivided patients into three risk groups according to these two factors. Systemic chemotherapy combined with LRRT may benefit patients in low- and intermediate-risk groups but not in the high-risk group. Further aggressive MRT based on systemic chemotherapy showed no survival benefit in any risk group. CONCLUSION: The stratification of NPC patients with bone-only metastasis based on EBV DNA after PCT and the number of metastatic lesions provided promising prognostic value and could aid clinicians in person-specific treatment.
Diagnosis ; DNA ; Drug Therapy ; Follow-Up Studies ; Herpesvirus 4, Human ; Humans ; Neoplasm Metastasis ; Prognosis ; Proportional Hazards Models ; Radiotherapy ; Retrospective Studies