Protein kinase D1 regulates the growth and metabolism of oral squamous carcinoma cells in tumor microenvironment / 华西口腔医学杂志

OBJECTIVE@#To observe the effect of protein kinase D1 (PKD1) on the growth and metabolism of oral squamous cell carcinoma HSC-4 cells and related molecular mechanisms in the tumor microenvironment.@*METHODS@#HSC-4 cell lines were transfected with shRNA plasmids. Three groups (Wild, control-shRNA, and PKD1-shRNA) were cultured under acidic or hypoxic environment for a certain time. Western blot was used to detect the expression of autophagy-related and glycolytic-related proteins. The proliferation changes were detected by CCK-8 kits.@*RESULTS@#The PKD1-knockdown HSC-4 cell line was established. PKD1 silencing increased autophagy activity. Under hypoxic and acidic conditions, the PKD1-knockdown HSC-4 cells showed lower proliferation than the parental cells. PKD1-knockdown also decreased the expression of hypoxia induciblefactor 1α (HIF-1α) and pyruvate kinase M2 (PKM2).@*CONCLUSIONS@#Under hypoxic and acidic conditions, PKD1 gene silencing can increase apoptotic autophagy activity. Downregulated PKD1 gene expression can reduce the glycolysis of oral squamous cell carcinoma cells and inhibit tumor cell proliferation. This study revealed the important role of PKD1 in the metabolism and growth of oral squamous cell carcinoma, making it a possible target for the treatment of oral squamous cell carcinoma.

Role of protein kinase D1 in regulating the growth, apoptosis and drug sensitivity of oral squamous carcinoma cells / 华西口腔医学杂志

OBJECTIVE@#This study aimed to investigate the role of protein kinase D (PKD)1 in regulating the growth, apop-tosis, and drug sensitivity of the squamous carcinoma cell line SCC-25.@*METHODS@#The SCC-25 cell line was transfected with either the control-shRNA or PKD1-shRNA plasmids. The stable transfected cells were selected, and the efficiency of PKD1 knockdown was detected by Western blot. The growth and apoptosis of SCC-25 were analyzed with a cell counting kit-8 (CCK8) and flow cytometry. The 50% inhibitory concentrations (IC50) of paclitaxel in the control and PKD1 knockdown cell lines were detected by CCK-8. The expression levels of Bax, Bcl-2, and P-gp were detected by Western blot.@*RESULTS@#PKD1 was constitutively expressed and phosphorylated in various cancer cell lines. Inhibiting the expression of PKD1 in SCC-25 cells by RNA interference could inhibit the growth and promote the apoptosis of SCC-25 cells via downregulating Bcl-2 expression. Additionally, inhibiting PKD1 expression could downregulate the expression of P-gp, thereby decreasing both the IC50 and resistance index of paclitaxel.@*CONCLUSIONS@#PKD1 plays an important role in regulating the biobehavior of SCC-25. It is a potential therapeutic target for oral squamous carcinoma.

Preparation of biotin-grafted pullulan acetate nanoparticles conjugated with CD3 antibodies and their uptake of T cells / 医学研究生学报

Objective Cytotoxic T lymphocytes are the main effector cells of anti-tumor immunity. Active targeting of nanoparticles to T cells and activation of T cells can be achieved by conjugation with specific antibodies. We prepared the biotin-grafted pullulan acetate nanoparticles conjugated with CD3 (Bio-PA-CD3 NPs), and explored their effects on the proliferation, cytokine secretion and uptake of CD8+T cells.Methods We prepared Bio-PA NPs by the dialysis method, conjugated CD3 antibodies to the surface of NPs to make Bio-PA-CD3 NPs, and measured the diameter and Zeta potential of the NPs. We evaluated the effects of the NPs on the proliferation of CD8+T cells and the secretion of cytokines by CCK-8 assay and ELISA, respectively, and quantitatively analyzed the cellular uptakes of the Bio-PA-CD3 NPs by the flow cytometry.Results The Bio-PA-CD3 NPs exhibited regular spherical shapes of even size and with no adhesion. The content of CD3 antibodies on the surface of the NPs decreased with the increased degree of biotin substitution. The CD3 contents of the Bio-PA-CD3 NPs with biotin substitution degrees of 1.6%, 5.4% and 6.3% were (36.1±4.4), (21.4±4.3) and (10.3±4.7) μg/mg, respectively. Compared with Bio-PA NPs, Bio-PA-CD3 NPs at a certain concentration significantly enhanced the proliferation of CD8+T cells in vitro and promoted the secretion of IFN-γ, TNF-β and IL-2 cytokines. The Bio-PA-CD3 NPs manifested a higher cellular uptake with the increased content of CD3 antibodies.Conclusion The Bio-PA-CD3 NPs we prepared could be a promising agent to enhance the immune effect of T cells.

Expression of miR-125a/b-5p in Serum and Urine of Preeclampsia and Related Clinical Studies / 现代检验医学杂志

Objective To detect miR-125a/b-5p expression in serum and urine of patients with preeclampsia,explore the role of miR-125a/b-5p in peripheral circulation on the pathogenesis of preeclampsia.Methods Samples of serum and urine of pa-tients with preeclampsia(n=20),and the normal pregnancy group(n=20)were collected and the expression of miR-125a/b-5p was detected by stem-loop-based real-time fluorescence quantitative reverse transcriptase polymerase chain reaction.Re-sults The expression of miR-125b-5p in the serum of patients with preeclampsia were significantly decreased compared with the normal pregnancy(Z=-2.272,P=0.023).There were no statistically differences of the expression of miR-125a-5p in serum between the two groups(Z=-0.622,P=0.547).The differences of urinary miR-125a/b-5p between patients with preeclampsia and normal pregnancy were not statistically significant(Z=-0.663,-0.189,P>0.05).Urinary miR-125b-5p was negatively correlated with diastolic blood pressure in preeclampsia group(r=-0.513,P=0.021).Serum miR-125b-5p was positively correlated with urea nitrogen in normal pregnancy group(r=0.472,P=0.036).Urinary miR-125a/b-5p were positively correlated with systolic blood pressure in normal pregnancy group(r=0.526,0.502,P=0.017,0.024). Conclusion The decrease of serum miR-125b-5p was associated with the pathogenesis of preeclampsia.

Research progress of structures and pharmacological activities of phthalides from Angelica sinensis / 中国中药杂志

Angelica sinensis(Umbelliferae)is a worldwide-known medicinal plant and also a famous traditional Chinese medicinal herb. It is recorded to possess the efficacy of enriching the blood and invigorating the circulation of blood of the individual.Danggui was extensively applied to the treatment of gynecological disorders. Modern researches indicate that phthalides are main chemical components related to the bioactivities of A.sinensis, such as anti-tumor, analgesic and neuroprotective effect.The advances in studies on the structures and pharmacological activities of phthalides from A.sinensis are reviewed to provide references for further researches and utilization of their medicinal value.

Zhibai Dihuang Decoction improves the activity of sperm mitochondrial respiratory chain complex in rats with Ureaplasma urealyticum infection / 中华男科学杂志

<p><b>Objective</b>To investigate the effect of Zhibai Dihuang Decoction (ZDD) on the sperm mitochondrial respiratory chain complex (MRCC) in rats with Ureaplasma urealyticum (UU) infection.</p><p><b>METHODS</b>Ninety male SD rats were randomly divided into five groups, sham operation, UU infection model control, ZDD (crude drug at 8.56 g per kg of the body weight per day), doxycycline (DC, at 20 mg per kg of the body weight per day), and ZDD+DC. The model of UU infection was established by injecting UU into the bladder of all the rats except those of the sham operation group. After modeling, the rats were treated intragastrically with respective drugs for 21 days and then executed and their epididymides harvested for examination of sperm quality and determination of the activities of sperm MRCCs I, II, III and IV by spectrophotometry.</p><p><b>RESULTS</b>At 10 days after modeling, the UU-positive rates in the model control, sham operation, ZDD, DC and ZDD+DC groups were 92.9%, 0%, 33.3%, 26.7% and 20.0%, respectively, significantly higher in the model control than in the other groups (P<0.05). The epididymal sperm concentrations in the five groups were (0.97±0.23), (3.02±0.52), (1.21±0.35), (1.02±0.31) and (1.52±0.28) ×106 ml, the sperm motilities were (58.62±15.36), (80.45±7.21), (75.52±8.78), (68.43±10.25) and (78.25±7.67)%, and rates of grade a+b sperm were (6.15±1.02), (10.32±1.14), (10.12±1.08), (9.01+1.27) and (10.74±1.03)%, respectively, all remarkably lower in the model control than in the sham operation group (P<0.01), but markedly higher in the ZDD and ZDD+DC groups than in the model controls (P<0.05). The activities of MRCC I in the model control, sham operation, ZDD, DC and ZDD+DC groups were (31.54±16.25), (136.86±6.34), (100.68±14.41), (81.68±6.78) and (124.06±5.54) μmol/(min·mg), those of MRCC II were (9.50±3.86), (20.34±0.37), (10.88±1.04), (12.93±1.07) and (16.23±0.60) μmol/(min·mg), those of MRCC III were (5.58±1.79), (19.60±0.61), (11.34±1.35), (13.87±1.23) and (15.96±0.69) μmol/(min·mg), and those of MRCC IV were (9.54±1.34), (28.98±3.33), (17.02±2.04), (18.41±2.67) and (21.66±2.93) μmol/(min·mg), respectively, all significantly lower in the model control than in the sham operation group (P<0.01), with the activities of MRCCs I, III and IV remarkably higher in the ZDD, DC and ZDD+DC groups (P<0.01) and that of MRCC II higher in the DC and ZDD+DC groups than in the model control (P<0.05).</p><p><b>CONCLUSIONS</b>ZDD can improve the epididymal sperm quality and the activity of the sperm MRCC in UU-infected rats, which may be one of the mechanisms of ZDD acting on male infertility caused by UU infection.</p>

Advances in the study of steroidal inhibitors of cytochrome P45017alpha / 药学学报

The steroidal enzyme cytochrome P45017alpha catalyzes the conversion of progesterone and pregnenolone into androgens, androstenedione and dehydroepiandrosterone, respectively, the direct precursors of estrogens and testosterone. Dihydrotestosterone is the principal active androgen in the prostate, testosterone is also an active stimulant of the growth of prostatic cancer tissue. Inhibition of this enzyme as a mechanism for inhibiting androgen biosynthesis could be a worthwhile therapeutic strategy for the treatment of PCA. In this paper, four categories of steroidal inhibitors of cytochrome P45017alpha will be reviewed, a diverse range of steroidal inhibitors had been synthesized and shown to be potent inhibitors of P45017alpha.

Vitro antitumor activity and synthesis of the key intermediate of bakuchiol / 药学学报

The in vitro antitumor activity of bakuchiol was exploited, compared with tamoxifen. The result of biological activities showed that bakuchiol could inhibit human breast cancer and the IC50 values were 2.89 x 10(-5) mol L(-1) and 8.29 x 10(-3) mol L(-1) against the cells line T-47D and MDA-MB-231 respectively. On the other hand, the key intermediate to synthesize bakuchiol was obtained by the method of Ireland-Claisen rearrangement. Comparing with traditional Claisen rearrangement, the reaction conditions are milder and the reaction reagents are safer.

Preparation and characterization of vincristine sulfate-loaded PLGA microspheres / 中国医学科学院学报

<p><b>OBJECTIVE</b>To prepare the vincristine sulfate (VCR) microspheres by W/O/O solvent evaparation method and evaluate the effect of zinc carbonate (ZnCO3) on the morphology and release kinetics of the microspheres.</p><p><b>METHODS</b>Degradation kinetic of VCR was tested in PBS of four different pH values at 37TC to select the optimal incubation medium for in vitro release. Microspheres were made with or without Zn-CO3 (w/w 5% and 10%) in the polymeric phase. The properties and in vitro release profiles of the microspheres were examed.</p><p><b>RESULTS</b>ZnCO3 increased the stability of VCR in the PLGA microspheres. During the 36 days of in vitro release, the accumulative release of VCR from the microspheres reached > 70% when added with ZnCO3, and was (54.2 +/- 1.1)% when no ZnCO3 was added. 10% ZnCO3 showed superior effect than 5% ZnCO3 in the stabilization of microspheres.</p><p><b>CONCLUSIONS</b>Adding ZnCO3 is essential during the preparation of PLGA microspheres. It can remarkably improve the stability of drugs in the acid microenvironment inside PL-GA microspheres and decrease the VCR degradation during incubation.</p>

Effect of phosgene on apoptosis of alveolar type II cells and vascular endothelial growth factor in exposed mice / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the apoptosis of alveolar type II cells, alterations of vascular endothelial growth factor (VEGF), VEGF receptor (Flt1) in serum and lung and expression of VEGF mRNA in lung in pulmonary edema mice induced by phosgene.</p><p><b>METHODS</b>Twenty-six BALB/C mice were randomly divided into 2 groups: control group, exposed group (13 mice in each group). Mice of exposed group were intoxicated by inhalation of phosgene 11.9 mg/L for 5 minutes. Mice of control group were treated as the same way by inhalation of air. Isolation of mice alveolus type II cells 4 h after intoxication was carried out to observe their apoptosis under electron microscope. Contents of VEGF and Flt1 in lung and serum by ELISA, and expression of VEGF mRNA were determined.</p><p><b>RESULTS</b>Alveolar type II cells were identified by tannic acid staining and electron microscopy. After exposed to 11.9 mg/L of phosgene for 5 minutes, the apoptotic body in alveolus type II cells was found in exposed group. The contents of VEGF in serum and lung and Flt1 in lung of exposed mice [(134.07 +/- 120.26), (477.76 +/- 98.06), (1,2818.48 +/- 2,304.15) pg/ml] were significantly lower than those of control group [(445.57 +/- 173.30), (1,026.87 +/- 474.56), (21,976.51 +/- 7,421.01) pg/ml, P < 0.05] but the content of Flt1 in serum [(2,369.56 +/- 381.70) pg/ml] was higher than that in control group [(1,898.00 +/- 453.69) pg/ml, P < 0.05]. The expression of VEGF mRNA in pulmonary edema mice was decreased.</p><p><b>CONCLUSION</b>Phosgene can induce apoptosis of alveolar type II cells, and decrease in the content of VEGF and Flt1, and expression of VEGF mRNA in lung.</p>

Effect of acute phosgene inhalation on antioxidant enzymes, nitric oxide and nitric-oxide synthase in rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the effect of acute phosgene inhalation on the antioxidant enzymes, nitric oxide (NO) and nitric oxide synthase (NOS) in rats.</p><p><b>METHODS</b>Phosgene was produced by decomposing bis (trichdomethyl) carbonate in the presence of N,N-dimethyl formamide. SD rats were randomly divided into two groups: control and phosgene exposure groups. In a special experimental device with equipment modulating the gas flow, phosgene exposed rats inhaled phosgene quantitatively for five minutes. Two hours later, all the rats were sacrificed and the ratio of wet weight to dried weight of lung (WW/DW) was calculated. Peripheral blood, serum and liver were collected to examine the activities of antioxidant enzymes including glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), NOS, and NO level. The total content of proteins were also determined.</p><p><b>RESULTS</b>The WW/DW ratio of lung in phosgene exposure group was much higher than that in control group (P < 0.01). The activities of GST in serum and liver of phosgene exposure group increased significantly (P < 0.05). The activities of SOD, CAT, GSHPx and NOS in serum or blood and liver of phosgene exposure group were also increased significantly (P < 0.05). But the content of NO was significantly decreased (P < 0.01).</p><p><b>CONCLUSION</b>Acute phosgene inhalation may cause a dramatically changes of several antioxidant enzyme activities, and acute injury of liver to some extent in rats. The latter is related to reactive oxygen species. But the elevation of antioxidant enzyme activities suggests that antioxidative treatment for acute phosgene poisoning should not be considered first.</p>