Protective effect of Ghrelin against oxidative stress induced by high glucose in human retinal pigment epithelium cells / 国际眼科杂志(Guoji Yanke Zazhi)

·AIM: To investigate the effect of Ghrelin on oxidative stress induced by high glucose in human retinal pigment epithelium (RPE) cells. ·METHODS: RPE cells were cultured and divided into the negative control group, high sugar group, Ghrelin low dose group ( 10-9mol/L ) and high dose group (10-6mol/L). Cells survival rate were detected by CCK-8 colorimetry, cells oxidative damage were observed by oxygen sensitive fluorescence probe H2DCFDA staning, changes of intracellular reactive oxygen species ( ROS ) were detected by H2DCFDA staining, super oxide dismutase ( SOD) activity and malondialdehyde ( MDA) content were detected by spectrophotometer colorimetry. ·RESULTS: CCK-8 results showed that RPE cells survival rate increased to 54.79%±3.43% and 79.16%±3.29% after treated with 10-9mol/L, 10-6mol/L Ghrelin, the difference was statistically significant compared with high glucose group (41.65%±3.42%)(P<0.05). H2DCFDA fluorescent probe dying showed that Ghrelin reduced ROS generation in RPE cells and decreased oxidative damage cells. Spectrophotometer colorimetric method showed that according to the high sugar group, SOD activity increased and MDA content decreased in Ghrelin group. ·CONCLUSION: Ghrelin could inhibit high glucose - induced oxidative damage in human RPE cells, which has protective effect on the process of the occurrence and development of diabetic retinopathy.

Investigation on the current situation of nursing specialty construction at department level in Shanghai tertiary hospitals / 中华护理杂志

Objective To investigate the current situation of nursing specialty construction at department level in Shanghai tertiary hospitals.Methods Totally 92 nursing specialties in 6 hospitals were investigated using a selfdesigned questionnaire of evaluation indicator system for nursing specialty construction at department level in tertiary hospitals.Results The score rate of 92 nursing specialties was between 27.75％～75.10％.There were 6 departments with a score of 70％ to 79.9％,accounting for 6.52％.There were 18 departments with a score of 60％ to 69.9％,accounting for 19.57％.There were 68 departments with a score less than 60％,accounting for 73.91％.For the first-level indicator,the score rate of "nursing practice" was the highest,followed by "teaching level""specialized training base construction""talent echelon construction" and "scientific research".Conclusion The overall level of nursing specialty construction at department level in Shanghai tertiary hospitals is relatively low.Hospitals should pay attention to "talent echelon construction","scientific research" and "training base construction" to increase the level of nursing specialist construction.

Kinematic analysis of a posterior-stabilized knee prosthesis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The goal of total knee arthroplasty (TKA) is to restore knee kinematics. Knee prosthesis design plays a very important role in successful restoration. Here, kinematics models of normal and prosthetic knees were created and validated using previously published data.</p><p><b>METHODS</b>Computed tomography and magnetic resonance imaging scans of a healthy, anticorrosive female cadaver were used to establish a model of the entire lower limbs, including the femur, tibia, patella, fibula, distal femur cartilage, and medial and lateral menisci, as well as the anterior cruciate, posterior cruciate, medial collateral, and lateral collateral ligaments. The data from the three-dimensional models of the normal knee joint and a posterior-stabilized (PS) knee prosthesis were imported into finite element analysis software to create the final kinematic model of the TKA prosthesis, which was then validated by comparison with a previous study. The displacement of the medial/lateral femur and the internal rotation angle of the tibia were analyzed during 0-135° flexion.</p><p><b>RESULTS</b>Both the output data trends and the measured values derived from the normal knee's kinematics model were very close to the results reported in a previous in vivo study, suggesting that this model can be used for further analyses. The PS knee prosthesis underwent an abnormal forward displacement compared with the normal knee and has insufficient, or insufficiently aggressive, "rollback" compared with the lateral femur of the normal knee. In addition, a certain degree of reverse rotation occurs during flexion of the PS knee prosthesis.</p><p><b>CONCLUSIONS</b>There were still several differences between the kinematics of the PS knee prosthesis and a normal knee, suggesting room for improving the design of the PS knee prosthesis. The abnormal kinematics during early flexion shows that the design of the articular surface played a vital role in improving the kinematics of the PS knee prosthesis.</p>

Fucoxanthin induces human MCF-7 breast cancer cells apoptosis via endoplasmic reticulum pathway / 中国药学杂志

OBJECTIVE: The antiproliferative effects and molecular mechanism of fucoxanthin were evaluated in human MCF-7 breast cancer cells. METHODS: Cell viability was assessed by MTT assay; cell cycle distribution, apoptosis and [Ca2+]i were measured by FACScan; the protein expression of caspase-4, caspase-7, caspase-9, Bcl-2, Bax, calpain and cytochrome C in the MCF-7 cells was evaluated by Western Blotting. RESULTS: Fucoxanthin exerted potent antiproliferative effects, induced [Ca2+]i overload and apoptosis in MCF-7 cells. Furthermore, fucoxanthin-mediated apoptosis was related with the activation of caspase-4, caspase-7, caspase-9, cytochrome C release from mitochondrion, up-regulation of Bax and calpain, and also down-regulation Bcl-2 in a dose-dependent manner. CONCLUSION: Fucoxanthin can induce MCF-7 cells apoptosis via Ca2+/calpain/caspase-4 pathway.

Molecular mechanism of ophiopogonin B induced cellular autophagy of human cervical cancer HeLa cells / 药学学报

This study is to investigate the antitumor activity of ophiopogonin B (OP-B). MTT assay, flow cytometric analysis, acridine orange staining, Lyso-Tracker Red staining and HeLa-GFP-LC3 transfect cells assay were used to detect the proliferation activity, apoptosis and autophagy of HeLa cells. The results showed that OP-B exerted potent antiproliferative activity on HeLa cells, the cell growth inhibition effect of OP-B was not due to apoptosis and OP-B could induce autophagy of HeLa cells. OP-B also induced the protein expression up-regulation of Beclin-1 and promoted LC3 I transformation LC3 II, which were representative proteins of autophagy. Furthermore, 3-MA, an inhibitor of autophagy, not only inhibited OP-B-mediated autophagy but also almost completely reversed the antiproliferative effect of OP-B, suggesting that the growth inhibition effect of OP-B was autophagy dependent. Western blotting demonstrated that OP-B inhibited the phosphorylation of Akt and its' downstream vital protein, such as mTOR and p70S6K. In addition, OP-B also induced the protein expression up-regulation of PTEN, which is a negative regulation protein for Akt/mTOR signaling pathway. However, OP-B did not affect the protein expression of total Akt. Collectively, the antitumor effects of OP-B were autophagy-dependent via repression Akt/mTOR signaling pathway. Therefore, OP-B is a prospective inhibitor of Akt/mTOR and may be used as an alternative compound to treat cervical carcinoma.

Synergistic antitumor effects of tanshinone II A in combination with cisplatin via apoptosis in the prostate cancer cells / 药学学报

Treatment with the combination of Chinese herbs and cytotoxic chemotherapies showed a higher survival rate in clinical trials. In this report, the results demonstrated that the tanshinone II A, a key component of Salvia miltiorrhiza bunge, when it is combined with the cytotoxic drug cisplatin showed synergistic antitumor effects on human prostate cancer PC3 cells and LNCaP cells in vitro. Antiproliferative effects were detected with MTT assay. Cell cycle distribution and apoptosis were detected by flow cytometer. Protein expression was detected by Western blotting. The intracellular concentration of cisplatin was detected by high performance liquid chromatography. The results demonstrated that tanshinone II A significantly enhanced the antiproliferative effects of cisplatin on human prostate cancer PC3 cells and LNCaP cells with the increase of the intracellular concentration of cisplatin. These effects were correlated with cell cycle arrested at S phase and cell apoptosis. The apoptosis might be achieved through death receptor pathway and mitochondrial pathway. Furthermore, the Bcl-2 family members were also involved in this apoptotic process. Collectively, these results indicated that the combination of tanshinone II A and cisplatin had a better treatment effect in vitro not only on androgen-dependent LNCaP cells but also on androgen-independent PC3 cells.

Synthesis and antitumor activity of fluoroquinolone C-3 heterocycles-oxadiazole derivatives (II) / 中国药学杂志

OBJECTIVE: To discover an efficient route for the conversion of an antibacterial fluoroquinolone to an antitumor one. METHODS: Cyclo-condensation of ofloxacin hydrazide 2 with aromatic carboxylic acids in POCl3 gave the corresponding oxadiazole derivatives 3a-3j, and their antitumor activity was evaluated by MTT assay. RESULTS: Ten title compunds were synthesized and showed potential antitumor activity. CONCLUSION: Heterocycles as isosteric replacement of carboxyl are warrant further development.

Part IV: Design, synthesis and antitumor activity of fluoroquinolone C-3 heterocycles: bis-oxadiazole methylsulfide derivatives derived from ciprofloxacin / 药学学报

To explore an efficient strategy for further development of anticancer fluoroquinolone candidates derived from ciprofloxacin, a heterocyclic ring as the bioisosteric replacement of C3 carboxyl group led to a key intermediate, oxadiazole thiol (5), which was further modified to the bis-oxadiazole methylsulfides (7a-7h) and the corresponding dimethylpiperazinium iodides (8a-8h), respectively. Structures were characterized by elemental analysis and spectra data, and their anticancer activities in vitro against CHO, HL60 and L1210 cancer cells were also evaluated by MTT assay. The preliminary results show that piperazinium compounds (8) possess more potent activity than that of corresponding free bases (7).

Role of dysregulation of Bim in resistance of melanoma cells to endoplasmic reticulum stress-induced apoptosis / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To establish a model of ER stress-induced apoptosis with tunicamycin and to examine whether Bim is dysregulated and its potential role in resistance of melanoma cells to apoptosis under endoplasmic reticulum (ER) stress.</p><p><b>METHODS</b>A model of ER stress-induced apoptosis was established with tunicamycin. Apoptotic cells were quantitated using the annexin V/propidium iodide method by flow cytometry. Hoechst staining was also used to confirm the apoptotic cell death. Western blotting was used to measure the activation of caspase-3 and -9, and the expression of Bim, GRP78, CHOP, and Foxo1 at the protein level. The expression of Bim, CHOP and Foxo1 at the mRNA level was quantitated by qPCR. The siRNA technique was used to inhibit the expression of Bim.</p><p><b>RESULTS</b>Treatment of the melanoma cells with tunicamycin did not induce significant apoptosis and activation of caspase cascade, whereas it caused marked activation of caspase-3 and -9, and apoptosis in HEK293 cells which were used as a control. With exposure to tunicamycin (3 µmol/L) for 12, 24, 36 hours the Bim protein levels were not increased in Mel-RM and MM200 cells. Its mRNA levels were 0.37 ± 0.05, 0.13 ± 0.02 and 0.02 ± 0.01 in Mel-RM cells, while 0.41 ± 0.06, 0.16 ± 0.04 and 0.21 ± 0.03 in MM200 cells, respectively. The expression of Bim mRNA was significantly reduced compared with that in the control groups of the two cell lines (P < 0.01). siRNA knockdown of Bim protected HEK293 cells against activation of caspase-3. The cell apoptosis of Bim siRNA group was (5.69 ± 0.38)%, significantly lower than that of the siRNA control group (40.32 ± 1.64)% and blank control group (35.46 ± 2.01)% (P < 0.01). In the melanoma cells after exposure to tunicamycin (3 µmol/L) for 6, 12, 24, and 36 hours the transcription factor CHOP at mRNA level were significantly increased and the expressions at protein level were also up-regulated. The expressions of another transcription factor Foxo1 at mRNA level significantly decreased and the expressions at protein level were down-regulated, too.</p><p><b>CONCLUSIONS</b>The lack of Bim up-regulation contributes to the resistance of melanoma cells to ER stress-induced apoptosis and may be a mechanism by which melanoma cells adapt to ER stress conditions. Transcription factors CHOP and Foxo1 may be responsible for the dysregulation of Bim in melanoma cells upon ER stress.</p>

Synthesis and antibacterial activity of imidazothiadiazoles and heterocyclic-amine Mannich-base hydrochloride / 药学学报

To optimize the synthetic method and antibacterial activity of fused heterocyclic thiadiazole compounds, cyclocondensation of 2-(4-methoxyphenyl)-5-amino-1,3,4-thiadiazole (2) with alpha-chloro-4-chloro acetophenone (3) resulted in a key intermediate (4), 6 -(4-chlorophenyl)-2-(4-methoxyphenyl)-imidazo-[2,1-b][1,3,4]thiadiazole, which was carried out an nucleophilic substitution with substituted piperazine to give the corresponding free bases of piperazine (5a-5c), then followed by Mannich reaction with heterocyclicamines and formaldehyde to yield the corresponding Mannich bases, (1a-11) as respective hydrochloride salts. The structures were confirmed by IR, 1H NMR, MS and elemental analysis and the antibacterial activities in vitro of fifteen newly synthesized compounds were also tested against Gram positive bacteria and Gram negative bacteria with the standard 2-fold agar dilution method. The antibacterial results showed that the introduction of a polar group resulted in the enhancement of antibacterial activity in vitro. Thus, the structures of these fused compounds could further be investigated.

Selective revealing of gap junction currents in single inspiratory tracheal motor neurons / 生理学报

Little is known about how gap junctions are involved in the respiratory-related or other types of physiological neuronal activity since physiologically active gap junction currents (GJCs) have never been characterized from single respiratory-related neurons or from single neurons of any other types. In the present study we hypothesized that GJCs could be selectively revealed from single neurons by elimination of transmembrane electrochemical gradients in voltage patch-clamp recording, and this hypothesis was tested in single inspiratory tracheal preganglionic vagal motor neurons (I-TPVMs). The results showed that GJCs were selectively revealed in all I-TPVMs when the transmembrane electrochemical gradients were eliminated in voltage patch-clamp recording, and were rhythmically activated by central inspiratory activity. Therefore, this method may be used as a fast way to detect GJCs within spontaneously active neuronal networks.