Objective To evaluate the diagnostic utility of repetitive nerve stimulation (RNS)and the effect of exercise test in RNS,as well as to find the way to improve the diagnostic sensitivity of RNS in myasthenia gravis (MG).Methods A total of 45 patients with generalized MG,admitted to our hospital from January 2008 to June 2012,were chosen in our study; they,firstly,underwent RNS test in orbiculatis otis,anconeus,deltoid and musculus abductor digiti minimi with low frequency (1,3 and 5Hz) and high frequency (10,20 and 30 Hz) supramaximal current,and then,till the muscle tetanic contraction (fatigue),RNS test at the same frequency was performed again; the diagnostic sensitivity of RNS in MG was compared.Results Before exercise test,the comprehensive positive rate of the four muscles on low-frequency RNS test was 60.0％ (27/45),which was significantly higher than that on high-frequency RNS test (17.78％,8/45) (x2=16.878,P=0.000).As compared with that before exercise test,the RNS positive rate of anconeus and deltoid after exercise was significantly higher (P＜0.05),while no significant difference was noted on orbicularis otis and abductor digiti minimi between before exercise test and after exercise test (P＞0.05).Increased comprehensive positive rate of four muscles in low-frequency RNS test was showed as compared with that in high-frequency RNS test (P＜0.05).Conclusion RNS has important diagnosis value in MG,and low-frequency RNS enjoys high positive rate; different muscles have different positive rate of RNS,and RNS for the four muscles at the same time can obviously increase the positive rate and detection sensitivity; exercise test can significantly improve the diagnostic sensitivity of RNS in anconeus and deltoid,as well as the comprehensive positive rate of four muscles in low-frequency RNS test.

Objective　To investigate the protective effect of calcium antagonist Verapamil (VP) on kidney preservation in HCA solution. Methods　After kidneys were isolated from rabbits, they were perfused and stored in HCA solution or in HCA solution with VP pre-supplement at 4℃ for 24 h respectively. The contents of mitochondrial calcium in renal cells and ATP in renal tissues were measured in every group. Results　The contents of mitochondrial calcium was remarkably higher and ATP significantly lower in the kidneys in HCA solution at 4℃ for 24 h than those just after resection. But these could be inhibited in those storing in the HCA solution with VP pre-supplement. Conclusion　Calcium antagonist VP can protect kidney function during HCA solution preservation by inhibiting calcium intaking into mitochondrium.

OBJECTIVETo investigate the mechanism by which heat shock protein 90 (HSP90) regulates 26S proteasome in hyperthermia.

METHODSHyperthermic HepG2 cell models established by exposure of the cells to 42 degrees celsius; for 3, 6, 12, and 24 h were examined for production of reactive oxygen species (ROS) and cell proliferation, and the changes in Hsp90α and 26S proteasome were analyzed.

RESULTSROS production in the cells increased significantly after hyperthermia (F=28.958, P<0.001), and the cell proliferation was suppressed progressively as the heat exposure time extended (F=621.704, P<0.001). Hyperthermia up-regulated Hsp90α but decreased the expression level (F=164.174, P<0.001) and activity (F=133.043, P<0.001) of 26S proteasome. The cells transfected with a small interfering RNA targeting Hsp90α also showed significantly decreased expression of 26S proteasome (F=180.231, P<0.001).

CONCLUSIONThe intracellular ROS production increases as the hyperthermia time extends. Heat stress and ROS together cause protein denature, leading to increased HSP90 consumption and further to HSP90 deficiency for maintaining 26S proteasome assembly and stability. The accumulation of denatured protein causes unfolded protein reaction in the cells to eventually result in cell death.

HSP90 Heat-Shock Proteins ; metabolism ; Hep G2 Cells ; Hot Temperature ; Humans ; Proteasome Endopeptidase Complex ; metabolism ; RNA, Small Interfering ; genetics ; Reactive Oxygen Species ; metabolism ; Up-Regulation

OBJECTIVETo study the effect of Erigeron Breviscapus (EB) at different concentrations and different intervention time points on the mRNA and protein expression of OPG/RANKL/RANK in MG63 osteoblast-like cells and RAW264. 7 pre-osteoclast cells cultured in vitro, thus exploring roles EB played in bone rebuilding and its mechanisms.

METHODSMG63 osteoblast-like cells and RAW264.7 pre-osteoclast cells were cultured in vitro. The 3rd passage cells were divided into the control group and different experimental groups. Total RNA and protein were respectively isolated from cells treated with different concentrations of EB (0, 0.001, 0.01, 0.1, and 1.0 mg/mL) for 48 h. Meanwhile, the protein was extracted from 0 and 1 mg/mL EB groups at 12, 24, and 48 h respectively. Expression of OPG mRNA and RANKL mRNA in MG63 osteoblast-like cells, and expression of RANK mRNA in RAW264.7 pre-osteoclast cells were detected by semi-quantitative RT-PCR. Expression of OPG protein and RANKL protein in MG63 osteoblast-like cells, and expression of RANK protein in RAW264. 7 pre-osteoclast cells were detected by Western blot.

RESULTSAlong with increased EB concentration, expression of OPG mRNA and protein in MG63 osteoblast-like cells was gradually lowered (P < 0.05) after 48-h intervention of EB, the expression of RANKL mRNA and protein in MG63 osteoblast-like gradually increased (P < 0.05); the expression of RANK mRNA in RAW264.7 pre-osteoclast cells increased (P < 0.05). But the expression of RANK mRNA was slightly lower in the 0.1 mg/mL EB group than in the 0.01 mg/mL EB group, and the expression of RANK protein in RAW264.7 pre-osteoclast cells gradually increased (P < 0.05). After treatment with 1 mg/mL EB for 12, 24, 48 h, the expression of OPG protein in MG63 osteoblast-like cells gradually decreased as time went by (P < 0.05), and the expression of RANKL protein in MG63 osteoblast-like and RANK protein in RAW264.7 pre-osteoclast cells gradually increased (P < 0.05). The expression of RANKL protein in RAW264.7 pre-osteoclast cells increased as time went by (P < 0.05).

CONCLUSIONEB could inhibit the expression of OPG in osteoblasts in a dose- and time-dependent manner, promote the expression of RANKL in osteoblasts and the secretion of RANK in pre-osteoclast, indicating EB might play roles in promoting bone resorption.

Animals ; Cell Differentiation ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Erigeron ; Humans ; Mice ; Osteoblasts ; drug effects ; metabolism ; Osteoclasts ; drug effects ; metabolism ; Osteoprotegerin ; metabolism ; RANK Ligand ; metabolism ; RNA, Messenger ; genetics ; Receptor Activator of Nuclear Factor-kappa B ; metabolism

In order to establish the quality standard of Berberidis Cortex and improve its quality control level, water, total ash, acid-insoluble ash and alcohol-soluble extract were determined according to procedures recorded in the Chinese Pharmacopoeia (2010 edition). The qualitative and quantitative analyses were performed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) methods. The results showed that TLC identification had a good resolution with clear spots. The water content was 8.39%-12.23%; total ash was 4.50%-9.96%; acid-insoluble ash was 0.10%-0.69%, and the alcohol-soluble extraction was 20.62%-37.13%. The average contents of magnoflorine, jatrorrhizine, palmatine, and berberine in Berberidis Cortex were 5.98%, 0.63%, 0.30%, 2.50%, respectively. It was concluded that the developed method was accurate and good in specificity, which can be used for quality control of Berberidis Cortex in the future.

OBJECTIVE: To study the in vitro and in vivo antitumor effects of the polysaccharide of Alocasia cucullata (PAC) and the underlying mechanism. METHODS: B16F10 and 4T1 cells were cultured with PAC of 40 µg/mL, and PAC was withdrawn after 40 days of administration. The cell viability was detected by cell counting kit-8. The expression of Bcl-2 and Caspase-3 proteins were detected by Western blot and the expressions of ERK1/2 mRNA were detected by quantitative real-time polymerase chain reaction (qRT-PCR). A mouse melanoma model was established to study the effect of PAC during long-time administration. Mice were divided into 3 treatment groups: control group treated with saline water, positive control group (LNT group) treated with lentinan at 100 mg/(kg·d), and PAC group treated with PAC at 120 mg/(kg·d). The pathological changes of tumor tissues were observed by hematoxylin-eosin staining. The apoptosis of tumor tissues was detected by TUNEL staining. Bcl-2 and Caspase-3 protein expressions were detected by immunohistochemistry, and the expressions of ERK1/2, JNK1 and p38 mRNA were detected by qRT-PCR. RESULTS: In vitro, no strong inhibitory effects of PAC were found in various tumor cells after 48 or 72 h of administration. Interestingly however, after 40 days of cultivation under PAC, an inhibitory effect on B16F10 cells was found. Correspondingly, the long-time administration of PAC led to downregulation of Bcl-2 protein (P<0.05), up-regulation of Caspase-3 protein (P<0.05) and ERK1 mRNA (P<0.05) in B16F10 cells. The above results were verified by in vivo experiments. In addition, viability of B16F10 cells under long-time administration culture in vitro decreased after drug withdrawal, and similar results were also observed in 4T1 cells. CONCLUSIONS Long-time administration of PAC can significantly inhibit viability and promote apoptosis of tumor cells, and had obvious antitumor effect in tumor-bearing mice.
Mice ; Animals ; Alocasia/metabolism* ; MAP Kinase Signaling System ; Caspase 3/metabolism* ; Apoptosis ; RNA, Messenger/metabolism*

Objective: Several COVID-19 patients have overlapping comorbidities. The independent role of each component contributing to the risk of COVID-19 is unknown, and how some non-cardiometabolic comorbidities affect the risk of COVID-19 remains unclear. Methods: A retrospective follow-up design was adopted. A total of 1,160 laboratory-confirmed patients were enrolled from nine provinces in China. Data on comorbidities were obtained from the patients' medical records. Multivariable logistic regression models were used to estimate the odds ratio ( Results: Overall, 158 (13.6%) patients were diagnosed with severe illness and 32 (2.7%) had unfavorable outcomes. Hypertension (2.87, 1.30-6.32), type 2 diabetes (T2DM) (3.57, 2.32-5.49), cardiovascular disease (CVD) (3.78, 1.81-7.89), fatty liver disease (7.53, 1.96-28.96), hyperlipidemia (2.15, 1.26-3.67), other lung diseases (6.00, 3.01-11.96), and electrolyte imbalance (10.40, 3.00-26.10) were independently linked to increased odds of being severely ill. T2DM (6.07, 2.89-12.75), CVD (8.47, 6.03-11.89), and electrolyte imbalance (19.44, 11.47-32.96) were also strong predictors of unfavorable outcomes. Women with comorbidities were more likely to have severe disease on admission (5.46, 3.25-9.19), while men with comorbidities were more likely to have unfavorable treatment outcomes (6.58, 1.46-29.64) within two weeks. Conclusion Besides hypertension, diabetes, and CVD, fatty liver disease, hyperlipidemia, other lung diseases, and electrolyte imbalance were independent risk factors for COVID-19 severity and poor treatment outcome. Women with comorbidities were more likely to have severe disease, while men with comorbidities were more likely to have unfavorable treatment outcomes.
Adult ; Aged ; COVID-19/virology* ; China/epidemiology* ; Comorbidity ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Severity of Illness Index ; Treatment Outcome