Objective To observe the enzymatic changes of myocardial sarcoplasmic reticulum in rats after injecting endotoxin (LPS), and provide basic research results for the future study of myocardial sarcoplasmic reticulum dysfunction caused by LPS in rats. Methods Ten SD rats were randomly divided into blank control group and LPS injection group with 5 rats in each group. In the LPS injection group, endotoxin was injected into the tail vessels of the rats. Results The heart rate (HR) of the LPS injection group increased and was faster than that in the blank control group [(204±18) beat per min vs. (139±10) beat per min on the first day, and (199±22) beat per min vs. (143±17) beat per min on the next day, both P values were less than 0.05]. The mean arterial pressure (MAP) was lower than that of the blank control group on the first day [(87±12) mmHg vs. (102±7) mmHg, P<0.05]. Under light and electron microscope, the myocardial cells of rats with LPS injection were loosely arranged, with intercellular infiltration with inflammatory cells, muscle fibers broken, and difficult to identify the morphology of mitochondria and sarcoplasmic reticulum. Quantitative PCR results showed that after endotoxin injection, troponin (CASQ1), sodium-calcium exchanger (NCX), calmodulin phosphatase 1 (ppplCa), phospholipid protein (PLN), sarcoplasmic reticulum Ca2+-ATPase (SERCA2) increased significantly (P<0.05). Conclusion Endotoxin can inhibit cardiomyocyte function by affecting the activity of sarcoplasmic reticulum calcium regulatory protein-related enzymes through various mechanisms.

Objective To characterize exosomes released from human umbilical vein endothelial cells (HUVECs) after heatstroke, and explore their effects on biological behaviors of cells. Methods HUVECs were cultured and divided into the control group and the heat shock group. The heat shock group cells were exposed at 41 ℃ for 2 hours; the control group cells were incubated at 37 ℃ for 2 hours. The characteristics of exosomes secreted by the two groups were analyzed by nanoparticle tracing. The expression of exosomes marker proteins, CD63 and TSG101, were detected by Western blotting. The process of exosomes entering normal cells was observed by fluorescence labeling. To further explore the potential function of exosomes secreted from the heat-shocked cells, cells were co-cultured with exosomes collected from cells cultured at 37 ℃ or cells cultured at 41 ℃. Cells under normal culture conditions were served as a negative control (blank control). FACS was used to detect cell cycle and apoptosis; MTS was used to evaluate cell proliferation; ELISA was used to quantify coagulation related factor expression; high-throughput sequencing was used to profile microRNAs (miRNAs) in the exosomes. Results Different from the control group, the cells in 41 ℃ heat shock group secreted abnormal exosomes, which resulted in abnormal behaviors and functions of normal HUVECs. The expression of tissue factor, von Willebrand factor, plasminogen activator inhibitor-1 increased significantly (P<0.05), and the expression of tissue plasminogen activator decreased (P<0.05). Compared with the incubation of HUVECs at 37 ℃, there were 2590 miRNA regulatory changes in exosomes secreted by HUVECs after heat shock at 41 ℃, which had obvious effects on apoptosis. Conclusions Heat shock at 41 ℃ could induce the release of abnormal exosomes from HUVEC, leading to abnormal behavior and potential coagulation of normal cells.

Objective To determine chromosomal localization of the primary gout susceptibility gene in a pedigree.Methods The clinical data and the peripheral blood samples were collected in the pedigree members and the genomic DNA was extracted from peripheral blood.A genome-wide screening was performed using 400 micro-satellite DNA markers in this family,and linkage analysis was used to determine the chromosomal location of the primary gout susceptibility gene.Results Linkage analysis showed that the maximum LOD score reached 1.50 at marker D4S1572 (at recombination fraction?=0.00).Conclusion Since D4S1572 is localized at 4q25,the primary gout susceptibility gene of this pedigree is localized at 4q25.

Objective To evaluate the effects of various polysorbates(PS)on the stability of different types of monoclonal antibody(mAb)drugs.Methods Three types of monoclonal antibodies mAbA(IgG1 proantibody drug),mAbB(IgG1 mAb)and mAbC(IgG1 mAb with Fc N297A mutation)were used as model proteins,and different kinds or contents of PS were added into the mAb formulations respectively to investigate the influencing factors.The effects of PS on the stability of mAb drugs were evaluated comprehensively by detecting the changes of quality attributes,such as protein aggregates and insoluble particles.Results PS20 and PS80 showed no significant difference in inhibiting the formation of aggregates and charge variants in the three mAbs(P>0.05),while the addition of PS80 in mAbB and PS20 in mAbC significantly inhibited the increase of insoluble particles respectively(P<0.05);The content of PS20 showed a significant effect on the detection indexes of charge variants and insoluble particles in mAbC(P<0.05).Conclusion Different types of mAbs have different sensitivities to various kinds and contents of PS.Therefore,when designing the formulation of mAbs,it is necessary to select appropriate kinds and contents of PS to further improve the stability of mAb drugs.

Objective To measure albumin in urine by HPLC and conduct primary clinical application Methods Solvent gradient and appropriate wave length was optimized and performance of the HPLC method was evaluated.Urine albumin of 46 patients with diabetes was measured.Results In standard and urine,retention time of Alb was 13.1 min.The linear measuring range extends to 1 820 mg/L.The lower limit of measurment for Alb was 4.2 mg/L.The intra-assay CV and the inter-assay CV were 3.36%,4.12% and 1.93%,1.97% at 24.5 mg/L and 546.9 mg/L of Alb respectively.Analytical recovery rate were 96.3%,98.2% and 97.5%.Microalbuminuria rate was 54.3% by HPLC,26.1% by immunoassay in 46 patients with diabetes.Conclusions Measurement of Alb in urine by HPLC is feasible as routine method until quantifying urinary total Alb conveniently.HPLC is the same to suit research for diabetic nephropathy and so on.

OBJECTIVETo detect the relationship between chromosomal anomalies and the pathogenesis, development and prognosis of ovarian carcinoma.

METHODSThirty-six specimens of ovarian carcinoma (n=12), ovarian benign tumor (n=12), and normal ovary (n=12) were examined by fluorescence in situ hybridization (FISH).

RESULTSTwelve cases of mutations, including trisomy 8, monosomy 8 or tetraploid 8 chromosomal anomalies, were found in the group of ovarian carcinoma, making up 100% (12/12). Three cases of trisomy 8 chromosomal anomalies were found in the group of ovarian benign tumor, accounting for 25% (3/12). No anomaly was found in the normal group. There were significant differences between the three groups, P<0.001.

CONCLUSIONThe above anomalies of chromosome 8 are significantly associated with the pathogenesis and development of ovarian carcinoma. The anomalies may occur in the early stage of the carcinoma, and may be significantly associated with the pathological differentiation and clinical stage of the case.

Adolescent ; Adult ; Aged ; Aneuploidy ; Chromosome Aberrations ; Chromosomes, Human, Pair 8 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Middle Aged ; Monosomy ; Ovarian Neoplasms ; genetics ; Trisomy

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Heart Atria ; pathology ; Histiocytoma, Malignant Fibrous ; diagnosis ; Humans ; Male ; Middle Aged ; Young Adult

Huangqi powder injection's positive rate of skin-test was 12.3%. Qingkailing powder injection was 3.0%. Qingkailing injection was 7.6%. Shuanghuanglian injection was 6.3%. Penicillin's rate of allergic reactions was 0.7%-10%. Different form of a drug (power or injection) and different drug consistency could influence the positive rate of skin-test. We don't use drug in positive group, and we use drug in negative group. Some trial subjects still happened allergic reactions in negative group of skin-test. In negative group of skin-test, Huangqi power injection's rate of allergic reactions was 2.1%. Qingkailing injection was 0.4%. Shuanghuanglian injection was 0.9%-2.6%. Shuanghuanglian injection's rate of allergic reactions was 8.6% in all subjects (31/360 include the subjects with positive skin-test and allergic reactions). Qingkailing powder injection's rate of allergic reactions was 4.5% (6/132). Qingkailing injection' s rate of allergic reactions was 9.1% (12/132). Huangqi power injection's rate of allergic reactions was 15.4% (62/402). The total rate of allergic reactions was 10.8%. The main appearance of Penicillin's skin-test was welling under skin, and with some blush. But the main appearance of traditional Chinese medicine skin-test was blush, and with a little welling under skin. Skin-test can reduce the allergic reactions of Qingkailing powder injection, Shuanghuangiian injection, Huangqi power injection. It can be the one measure of reducing adverse reactions.
Astragalus membranaceus ; adverse effects ; Drug Hypersensitivity ; etiology ; Drugs, Chinese Herbal ; adverse effects ; Humans ; Medicine, Chinese Traditional ; Polysaccharides ; adverse effects ; isolation & purification ; Skin Tests

OBJECTIVETo investigate the effect of nucleolin silencing on the differentiation of rat neural stem cells (NSCs) and the role of Wnt signaling pathway in mediating such effect.

METHODSAdenovirus vectors expressing small interfering RNA (siRNA) against nucleolin were constructed, verified, and packaged in HEK293A cells. The adenovirus was then transfected into NSCs isolated from neonatal SD rats and the differentiation of the NSCs was examined by detecting the expressions of neuron specific encloase (NSE) and glial fibrillary acidic protein (GFAP) using immunocytochemistry. The expressions of nucleolin, nestin, Wnt3, and β-catenin in the cells were determined with Western blotting.

RESULTSRestriction endonuclease and sequencing analysis verified successful construction of the adenoviral vector expressing nucleolin siRNA (nucleolin-siRNA2). Infection of rat NSCs with nucleolin-siRNA2 significantly lowered nucleolin protein expression as compared with that in negative and blank control groups (P<0.05). The percentages of NSE-positive cells and GFAP-positive cells were significantly higher in NSCs infected with nucleolin-siRNA (P<0.01); the infection also resulted in obviously lowered expression of nestin protein and increased expressions of Wnt3 protein and β-catenin nucleoprotein in the cells.

CONCLUSIONSNucleolin silencing by adenovirus-mediated RNA interference induces the differentiation of NSCs into neurons and astrocytes, which is related with the activation of Wnt signaling pathway.

OBJECTIVETo study the relation of the sex, age, location and chemotherapy with recurrence of the tumor.

METHODSFrom January 2000 to August 2010, 47 patients with giant cell tumor of tendon sheath in upper extremity were retrospectively analyzed. Statistical analysis of sex, age at presentation, lesion location, chemical inactivation, surgical complications, tumor recurrence and pathological findings were explored. There were 28 females and 19 males, ranging in age from 17 to 78 years, with an average of 38.15 years. All the patients underwent surgical excision. Fourteen patients received intraoperative chemically inactive treatment. All the patients had routine follow-up to observe the wound healing, pathological findings,tumor recurrence, and received necessary imaging examinations.

RESULTSAll the patients were followed up, and the duration ranged from 22 to 129 months, with a mean time of 53.89 months. Four patients who received intraoperative alcohol inactivation appeared wound complications such as wound swelling, discharge of necrotic tissue, delayed wound healing. Fifteen patients had active growth of tumor tissue, 1 patient had low-grade malignant giant cell tumor of tendon sheath. The recurrence rate was significantly higher in the group which preoperative X-ray was found to have bone destruction (P = 0.003); patients receiving chemically inactivation had lower risk of recurrence after surgery than patients not receiving chemically inactivation (P = 0.042).

CONCLUSIONThe recurrence rate of giant cell tumor of tendon sheath in upper limb was closely related to tumor growth site, bone destruction and chemical inactivation. Local excision of giant cell tumor of tendon sheath was the effective treatment. How to identify the patients at high risk of recurrence, how to reduce the recurrence rate and the functional restoration after wide resection are the priorities and difficulties of future researches.

Adult ; Female ; Giant Cell Tumors ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; epidemiology ; Postoperative Complications ; epidemiology ; Retrospective Studies ; Soft Tissue Neoplasms ; pathology ; surgery ; Tendons ; pathology ; Upper Extremity