Remifentanil Down-regulates GIRK2 Expression in Rat Dorsal Root Ganglion and Spinal Dorsal Horn / 中山大学学报(医学科学版)

ObjectiveTo observe the changes in the expression and distribution of G protein-gated inwardly rectifying potassium channel subunit 2 （GIRK2） in the dorsal root ganglion （DRG） and spinal cord dorsal horn of rats with remifentanil-induced hyperalgesia. MethodsHyperalgesia was induced by intravenous infusion of remifentanil 4 μg/kg/min for 2 h in adult male SD rats. At 6th hour and on days 1， 3 and 5 following remifentanil treatment， we used immunofluorescence to examine the changes in the GIRK2 distribution and expression. Immunoblotting was used to detect GIRK2 expression of the total protein and membrane protein in DRG and spinal dorsal horn of rats. Behavioral testing was applied to evaluate the effect of intrathecal injection of GIRK2-specific agonist ML297 on thermal nociceptive threshold on day 1 after remifentanil infusion. Resultsmmunofluorescence results showed that GIRK2 was mainly co-localized with IB4-positive small neurons in DRG and nerve fibers in spinal dorsal horn. GIRK2 expression was significantly downregulated following remifentanil treatment. Immunoblotting results revealed that on day 1 following intravenous infusion of remifentanil， compared with those in the control group， GIRK2 expression levels of the total protein and membrane protein in DRG （0.47 ± 0.10 vs. 1.01 ± 0.17， P < 0.001； 0.47 ± 0.11 vs. 1.06 ± 0.12， P < 0.001） and spinal dorsal horn （0.52 ± 0.09 vs. 1.10 ± 0.08， P < 0.001； 0.54 ± 0.10 vs. 1.01 ± 0.13， P < 0.001） were all significantly decreased. The behavioral results showed that intrathecal ML297 effect on thermal withdrawal latency was significantly reduced following remifentanil treatment （P < 0.001）. ConclusionsRemifentanil might induce hyperalgesia via down-regulating GIRK2 expression in rat DRG and spinal cord dorsal horn.

Risk factors of neovascular glaucoma after pars plana vitrectomy for proliferative diabetic retinopathy / 中华眼底病杂志

Objective:To analyze the risk factors of neovascular glaucoma (NVG) after 25G pars plana vitrectomy (PPV) in proliferative diabetic retinopathy (PDR).Methods:A retrospective study. From January 2017 to December 2018, 340 PDR patients (340 eyes) with vitreous hemorrhage (VH) who were first treated with PPV in Tianjin Medical University Eye Hospital were included in the study. Among them, 185 were male and 155 were female, with an average age of 55.79±10.82 years. The duration of diabetes was 13.01±7.70 years, the fasting blood glucose was 7.55±2.15 mmol/L. Nineteen patients combined coronary heart disease, and 20 patients combined cerebral infarction. All patients underwent best-corrected visual acuity (BCVA), intraocular pressure (IOP), non-contact fundus examination, and fundus color photographs. BCVA was measured using an international standard Snellen visual acuity chart, and the values were converted to logarithm of the minimum angle of resolution (logMAR) scores for data analysis. The baseline logMAR BCVA was 2.04±0.73, The baseline IOP was 15.45±2.93 mmHg (1 mmHg=0.133 kPa). The duration of VH was 2.98±1.46 months, ranged from 3 weeks to 6 months. Three hundred and forty eyes included 93 eyes of PDR Ⅳ stage (27.35%), 107 eyes of Ⅴ stage (31.47%), and 116 eyes of Ⅵ stage (34.12%), combined tractional retinal detachment (TRD) 83 eyes. All patients underwent 25G standard three channel vitrectomy through the pars plana of the ciliary body. Preoperative anti-VEGF injection was performed in 57 eyes, internal limiting membrane (ILM) peeling in 234 eyes, combined phacoemulsification cataract surgery in 262 eyes and 141 eyes intravitreal anti-VEGF injection at the end of surgery. The patients were followed up for at least 12 months, with an average follow-up time of 10.80±5.79 months. NVG was defined as the presence of neovascularization in the anterior chamber angle or iris with an IOP higher than 21 mmHg after vitrectomy. Kaplan-Meier method and Cox univariate and multivariate regression were used to analyze the relationship between baseline factors, ocular factors, surgical factors and the occurrence of NVG after surgery.Results:Among 340 eyes, 66 eyes (19.41%) developed NVG after vitrectomy during 12 months of observation, NVG occurred from 6 to 335 days after surgery, and the mean period between vitrectomy and developing NVG was 98.00±5.79 days. The incidence of NVG was 11.50%, 15.29% and 20.75%, respectively in the 3rd, 6th and 12th month after PPV. The result of univariate analysis with the Cox regression analysis showed that the development of NVG at 12 months after surgery and age, combined coronary heart disease or cerebral infarction, combined with cataract phacoemulsification, ILM peeling, preoperative anti-VEGF injection had effect on postoperative NVG ( P<0.05). Ocular factors such as PDR staging, combined TRD, preoperative logMAR BCVA, preoperative intraocular pressure, etc. had no effect on the occurrence of NVG after surgery ( P>0.05). Combined cataract phacoemulsification surgery, internal limiting membrane peeling, surgical factors such as intracavity injection of anti-VEGF drugs 3 days before surgery, had an impact on the occurrence of NVG after surgery ( P<0.05). The meaningful variables of the Cox univariate analysis were incorporated into the multivariate Cox proportional hazard model for analysis, and the influencing factors of NVG after surgery were gradually regressed. The results showed that age, coronary heart disease or cerebral infarction, combined with phacoemulsification of cataract, and internal limiting membrane removal during surgery were independent risk predictors of NVG after surgery ( P<0.05). Conclusions:Younger, coronary heart disease or cerebral infarction, combined with cataract phacoemulsification are the risk factors of NVG in PDR patients after PPV. The removal of internal limiting membrane can reduce the incidence of NVG.

Involvement of Spinal CCL2 in Remifentanil-induced Hyperalgesia in Rats / 中山大学学报(医学科学版)

remifentanil；hyperalgesia；CCL2；microglia@#【Objective】To observe C-C motif chemokine ligand 2（CCL2）mediates remifentanil-induced hyperalgesia by activating microglia in rats.【Methods】Thirty-six adult male SD rats were equally and randomly divided into 6 group with 6 rats in each group：normal saline ，remifentanil，saline + intrathecal injection of phosphate buffer saline（PBS）， remifentanil + intrathecal injection of PBS，saline + intrathecal injection of CCL2 neutralizing antibody and remifentanil + CCL2 neutralizing antibody. To establish remifentanil-induced hyperalgesia model，remifentanil［4 μg/ （kg·min）2 h］ was continuously infused into rats via the tail vein. Then ，immunofluorescence and western blot were performed to examine the expression of spinal CCL2 in rats. Thirty minutes before remifentanil infusion，intrathecal injection of CCL2 neutralizing antibody was performed to observe remifentanil-induced hyperalgesia and the effect on the activation of microglial in the spinal cord of rats.【Results】Compared with the normal saline group，the thermal and mechanical pain thresholds in the remifentanil group were significantly reduced（P < 0.05）. The expression of protein（0.66 ± 0.071 vs. 0.18 ± 0.04；P < 0.001）and average immunofluorescence density of spinal CCL2（0.170 ± 0.009 vs. 0.047 ± 0.006；P < 0.001）were significantly increased on the first day after remifentanil infusion. Prior intrathecal injection of CCL2 neu⁃ tralizing antibody not only inhibited remifentanil-induced activation of microglia in the spinal cord（P < 0.001），but also mitigated remifentanil-induced thermal hyperalgesia and mechanical pain sensitivity（P < 0.05）.【Conclusion】CCL2 me⁃ diates the remifentanil-induced hyperalgesia by activating spinal microglia in rats.

In Vitro Evaluation of Poloxamer 188 Modified Polyethyleneimine as Gene Vectors / 医药导报

Objective To modify polyethyleneimine (PEI) by using Poloxamer 188 (P188) , and evaluate its related feature as carriers of genes in vitro. Methods PEI was modified through conjugating one hydroxyl group of P188 to the amino group of PEI by carbonate method. Structural analysis of synthesized polymer was performed by using 1H-NMR. The particle size and Zeta potential of synthesized polymer /DNA complexes were measured. The cytotoxicity of the complexes was evaluated using MTT method in MCF-7 cells, HeLa cells and HepG2 cells. The pGL3-lus was used as a reporter gene, and the transfection efficiency of complexesat HeLa cells was evalutated by measuring activity of luciferase. Results The result of 1H-NMR showed the purity of these synthesized polymers was high. The particle size of complexes were decreased with the increment of N /P ratios. The Zeta potential of complexes increased with the increment of N /P ratios. The cytotoxicity of the complexes increased with the increment of N /P ratios. The synthesized polymers showed lower cytotoxicity than unmodified PEI. The new synthesized polymers had maintained the high transfection efficiency at high N /P ratios. In particular, the optimal transfection efficiency of (P188) 1- PEI (N /P = 24) was significantly higher than that of PEI (N /P = 6) . Conclusion The P188 modifed PEI can serve as a effective non-viral gene carriers to transfect Hela cells.

Improvement of the Synthesis Technology of 7-methoxy-4-(2-methyl-4-quinazolinyl)-3,4-dihydroquinoxalin-2 (1H)-ketone / 中国药房

OBJECTIVE:To improve the synthesis technology of 7-methoxy-4-(2-methyl-4-quinazolinyl)-3,4-dihydroquinoxalin-2 (1H)-ketone. METHODS:Using 2-methyl-4(3H)-quinazolone as starting material,the synthesis technology of 7-methoxy-4-(2-methyl-4-quinazolinyl)-3,4-dihydroquinoxalin-2(1H)-ketone was improved by clorination, nucleophilic substitution, diarylamine alkylation and nitro reductive cyclization. The yield of it was investigated. RESULTS:The structure of 7-methoxy-4-(2-methyl-4-quinazolinyl)-3,4-dihydroquinoxalin-2(1H)-ketone had been verified by 1H-NMR and ESI-MS. The yield was 43.5%and improved by 23.3% compared to 20.2% before improvement. CONCLUSIONS:Improved technology is simpler and milder, which is suitable for the batch preparation of laboratory study.

Progress in epigenetic study of clear cell renal cell carcinoma / 中国病理生理杂志

The epigenetic changes of clear cell renal cell carcinoma(ccRCC )are considered to be the main molecular mechanisms of its pathogenesis ,including DNA methylation ,histone modification ,microRNA change ,and so on.DNA methyltransferases(DNMTs)catalyze the occurrence of DNA methylation.DNA methylation changes are mani-fested in the overall low methylation of the genome and the high methylation of specific sites ,which were involved in the de-velopment of ccRCC by affecting the expression of tumor suppressor genes.Due to histone-modified enzyme involvement ,histone modification is shown as possible genetic reversal.MicroRNA plays an important role in the abnormal expression of ccRCC genes.With the studies of epigenetic mechanism and molecular pathology ,it is important to explore the mechanisms and to seek effective early diagnosis ,treatment and prognosis intervention of ccRCC.

Variations in cadmium and nitrate co-accumulation among water spinach genotypes and implications for screening safe genotypes for human consumption / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Vegetables are important constituents of the human diet. Heavy metals and nitrate are among the major contaminants of vegetables. Consumption of vegetables and fruits with accumulated heavy metals and nitrate has the potential to damage different body organs leading to unwanted effects. Breeding vegetables with low heavy metal and nitrate contaminants is a cost-effective approach. We investigated 38 water spinach genotypes for low Cd and nitrate co-accumulation. Four genotypes, i.e. JXDY, GZQL, XGDB, and B888, were found to have low co-accumulation of Cd (<0.71 mg/kg dry weight) and nitrate (<3100 mg/kg fresh weight) in the edible parts when grown in soils with moderate contamination of both Cd (1.10 mg/kg) and nitrate (235.2 mg/kg). These genotypes should be appropriate with minimized risk to humans who consume them. The Cd levels in the edible parts of water spinach were positively correlated with the concentration of Pb or Zn, but Cd, Pb, or Zn was negatively correlated with P concentration. These results indicate that these three heavy metals may be absorbed into the plant in similar proportions or in combination, minimizing the influx to aerial parts. Increasing P fertilizer application rates appears to prevent heavy metal and nitrate translocation to shoot tissues and the edible parts of water spinach on co-contaminated soils.

Minocycline inhibits formalin-induced inflammatory pain and the underlying mechanism / 北京大学学报(医学版)

OBJECTIVE@#To unravel the underlying mechanism of minocycline in formalin-induced inflammatory pain, and to investigate the effects of minocycline on synaptic transmission in substantia gela-tinosa (SG) neurons of rat spinal dorsal horn.@*METHODS@#Behavioral and immunohistochemistry experiments: 30 male Sprague-Dawley (SD) rats (3-5 weeks old) were randomly assigned to control (n=8 rats), model (n=8 rats), saline treatment model (n=6 rats) and minocycline treatment model (n=8 rats) groups. The control group was subcutaneously injected with normal saline on the right hindpaws. Acute inflammatory pain model was established by injecting 5% (volume fraction) formalin into the right hindpaws. The rats in the latter two groups received intraperitoneal injection of saline and minocycline 1 h before the formalin injection, respectively. The time of licking and lifting was recorded every 5 min within 1 h after the subcutaneous injection of normal saline or formalin for all the groups, which was continuously recorded for 1 h. One hour after the pain behavioral recording, the spinal cord tissue was removed following transcardial perfusion of 4% paraformaldehyde. The expression of c-Fos protein in spinal dorsal horn was observed by immunohistochemistry. Electrophysiological experiment: In vitro whole-cell patch-clamp recordings were performed in spinal cord parasagittal slices obtained from 26 male SD rats (3-5 weeks old). Two to five neurons were randomly selected from each rat for patch-clamp recording. the effects of minocycline, fluorocitrate and doxycycline on spontaneous excitatory postsynaptic currents (sEPSCs) or spontaneous inhibitory postsynaptic currents (sIPSCs) of SG neurons were investigated.@*RESULTS@#Compared with the control group, both the licking and lifting time and the expression of c-Fos protein in ipsilateral spinal dorsal horn of the model group were significantly increased. Intraperitoneal injection of minocycline largely attenuated the second phase of formalin-induced pain responses (t=2.957, P<0.05). Moreover, c-Fos protein expression was also dramatically reduced in both the superficial lamina (I-II) and deep lamina (III-IV) of spinal dorsal horn (tI-II=3.912, tIII-IV=2.630, P<0.05). On the other side, bath application of minocycline significantly increased the sIPSCs frequency to 220%±10% (P<0.05) of the control but did not affect the frequency (100%±1%, t=0.112, P=0.951) and amplitude (98%±1%, t=0.273, P=0.167) of sEPSCs and the amplitude (105%±3%, t=0.568, P=0.058) of sIPSCs. However, fluorocitrate and doxycycline had no effect on the frequency [(99%±1%, t=0.366, P=0.099); (102%±1%, t=0.184, P=0.146), respectively] and amplitude [(98%±1%, t=0.208, P=0.253); (99%±1%, t=0.129, P=0.552), respectively] of sIPSCs.@*CONCLUSION@#Minocycline can inhibit formalin-induced inflammatory pain and the expression of c-Fos protein in spinal dorsal horn. These effects are probably due to its enhancement in inhibitory synaptic transmission of SG neurons but not its effect on microglial activation or antibiotic action.

Seminal plasma anti-Müllerian hormone and inhibin B and serum inhibin B in predicting the outcome of routine IVF fertilization / 中华男科学杂志

Objective@#To analyze the correlations of seminal plasma (sp) anti-Müllerian hormone (spAMH) and inhibin B (spINHB) and serum INHB (serINHB) with semen parameters in oligoasthenospermia patients and explore their value in predicting the outcome of routine in vitro fertilization (IVF).@*METHODS@#We obtained the levels of spAMH, spINHB and serINHB as well as semen parameters from 88 infertile males undergoing IVF due to oligoasthenospermia or female uterine tubal factors from August 2016 to February 2017. Using the ROC curve and Pearson's correlation analysis, we examined the effects of the obtained parameters on the fertilization rate and assessed the correlation of the levels of spAMH, spINHB and serINHB with the semen parameters of the patients.@*RESULTS@#Concerning the predictive value for the outcome of IVF, Pearson's correlation analysis showed that the area under the ROC curve (AUC) of spAMH was 0.807 (sensitivity = 84.6%, specificity = 76%, cut-off point = 3.529, P <0.001) and that of spINHB was 0.768 (sensitivity = 84.6%, specificity = 88.7%, cut-off point = 31.117, P = 0.002). The serINHB level was found positively correlated with sperm concentration (r = 0.346, P = 0.001), total sperm count (r = 0.378, P <0.001), sperm motility (r = 0.521, P <0.001), and the percentage of progressively motile sperm (r = 0.343, P = 0.001).@*CONCLUSIONS@#The levels of spAMH and spINHB can be used as laboratory indexes to predict the fertilization rate of routine IVF and are correlated with semen parameters in oligoasthenospermia patients, while that of serINHB has a positive correlation with the semen parameters of the patients.

Preparation of a polyclonal antibody against human LYZL4 and its expression in the testis / 中华男科学杂志

Objective@#To prepare a polyclonal antibody against human lysozyme-like protein 4 (LYZL4) expressed in the prokaryotic system and identify the distribution of LYZL4 in the testis.@*METHODS@#The full-length cDNA of LYZL4 was cloned into the pET32a plasmid and the expression of the recombinant LYZL4 (rLYZL4) was induced by IPTG. The rLYZL4 was purified by Ni-NTA and chitin affinity chromatography respectively and its bactericidal activity was observed by bilayer agar plate diffusion assay. The purified rLYZL4 was used as an immunogen to generate the polyclonal antibody, followed by examination of the antibody titer by ELISA and its specificity by Western blot. The distribution of LYZL4 in human tissue, sperm and seminal plasma was identified and its subcellular localization in the testis was determined by immunohistochemistry.@*RESULTS@#rLYZL4 was expressed efficiently in the prokaryotic system and exhibited no bacteriolytic activity against M. lysodeikticus and E. coli. The anti-rLYZL4 polyclonal antibody could bind the recombinant protein with a high sensitivity and specificity. LYZL4 was identified in the testis, epididymis and sperm protein extracts and localized in the acrosomal region of round and elongating spermatids.@*CONCLUSIONS@#An anti-rLYZL4 polyclonal antibody was successfully prepared using the prokaryotic expression system. LYZL4 was detected in the acrosomal region of round and elongating spermatids, suggesting an association with the structure and function of the acrosome.

Identification of interacting proteins with NF-κB in different status of uterine smooth muscle in labor / 中南大学学报(医学版)

To analyze the differentially expressed proteins which interacted with NF-kappaB in the uterine lower segment smooth muscle tissues under different status of labor onset, and to provide a new foundation on the mechanisms for labor onset.
 Methods: NF-κB P65 protein expression in smooth muscle tissues from the term non-labor group, natural term labor group and drug-induced term labor group was analyzed by Western blot. Co-immunoprecipitation and SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) were performed to detect the proteins interacting with NF-κB p65 in the NF-κB p65 complexes. The components of the complex were identified by LC-ESI-MS/MS (liquid chromatography-tandem electrospray mass spectrometry) and database analysis. The identified differentially expressed proteins were confirmed by Western blot.
 Results: Positive expression of NF-κB was detected in all of the three groups. 10 differentially expressed proteins were identified by LC-ESI-MS/MS in human lower segment myometrium tissues in the term non-labor group and natural term labor group, mean while, 5 differentially expressed proteins were identified in the term non-labor group and the drug-induced labor group. 3 differential expression proteins were detected in all of the 3 groups, including Heat shock 70, Annexin A6 and Desmin, which were verified by Western blot. These proteins were mainly involved in chaperone, signal transduction, cell structure, and energy metabolism process, respectively.
 Conclusion: NF-κB expressed in uterine smooth muscle cells is involved in the process of initiation and regulation of labor onset through a number of proteins relevant to signal transduction, cell structure and energy metabolism.

Study on the regulation of autophagy against anticancer drugs' toxicity / 药学学报

Autophagy is a crucial biological process in eukaryotes, which is involved in cell growth, survival and energy metabolism. It has been confirmed that autophagy mediates toxicity of anticancer drugs, especially in heart, liver and neuron. It is important to understand the function and mechanism of autophagy in anticancer drugs-induced toxicity. Given that autophagy is a double-edged sword in the maintenance of the function of heart, liver and neuron, the autophagy-mediated toxicity are very complicated in the body. We provide a review on the concept of autophagy and current status about autophagy-mediated toxicity of anticancer drugs. The knowledge is crucial in the basic study of anticancer drugs-induced toxicity, and provides some strategies for the development of alleviating the toxicity of anticancer drugs.

Study on the regulation of autophagy against anticancer drugs' toxicity / 药学学报

Autophagy is a crucial biological process in eukaryotes, which is involved in cell growth, survival and energy metabolism. It has been confirmed that autophagy mediates toxicity of anticancer drugs, especially in heart, liver and neuron. It is important to understand the function and mechanism of autophagy in anticancer drugs-induced toxicity. Given that autophagy is a double-edged sword in the maintenance of the function of heart, liver and neuron, the autophagy-mediated toxicity are very complicated in the body. We provide a review on the concept of autophagy and current status about autophagy-mediated toxicity of anticancer drugs. The knowledge is crucial in the basic study of anticancer drugs-induced toxicity, and provides some strategies for the development of alleviating the toxicity of anticancer drugs.

Expression of human lysozyme-like protein 6 in the male reproductive system / 中华男科学杂志

<p><b>Objective</b>To study the expression of human lysozyme-like protein 6 (LYZL6) in the male reproductive system and its physiological role.</p><p><b>METHODS</b>The recombinant P. pastoris strain was cultured and induced with methanol to express LYZL6, followed by purification using chitin affinity chromatography. The bactericidal activity of the recombinant LYZL6 was observed by bilayer agar plate diffusion assay, and then the recombinant protein was used as an immunogen to generate polyclonal antibodies, whose specificity was examined by ELISA. The distribution of LYZL6 in the human tissue and semen was identified by Western blotting and the subcellular localization in the testis was investigated by immunohistochemistry.</p><p><b>RESULTS</b>At pH 5.6, recombinant LYZL6 exhibited a high bacteriolytic activity against M. lysodeikticus. ELISA analysis showed that the anti-LYZL6 polyclonal antibodies could bind the recombinant protein with a high specificity. Western blot manifested the expression of LYZL6 in the testis and epididymis, higher in the former than in the latter. LYZL6 was also detected in the sperm protein extract, while protein bands were not observed in the seminal plasma. Immunodetection with a specific antiserum localized the LYZL6 protein in the late spermatocytes and round spermatids.</p><p><b>CONCLUSIONS</b>LYZL6 has a higher bacteriolytic activity under low pH condition and is bound to spermatozoa after secreted in the testicular epithelia, suggesting that LYZL6 could act as a potential hydrolase for carbohydrates in zona pellucida penetration.</p>

Effects of negative capacity in the prevention of subcutaneous fluid accumulation after modified radical mas-tectomy / 中国基层医药

Objective To investigate the effect of negative capacity on drainage time,subcutaneous fluid accumulation,etc.after modified radical mastectomy.Methods 80 breast cancer patients after modified radical mas-tectomy.Used computer to produce random number,then 80 patients were randomly divided into the two groups.A group were adopted negative capacity of 200mL and B group adopted 400mL.Both groups used disposable negative pressure drainage balls.Results A group patients′subcutaneous fluid accumulation incidence was 27.91%,skin flap necrosis incidence was 2.33%,indwelling time was (25.00 ±2.59)days.B group patients′subcutaneous fluid accumulation incidence was 8.11%,skin flap necrosis incidence was 0,indwelling time was (12.00 ±2.34)days. Drainage time and subcutaneous fluid accumulation incidence were reduced in B group in comparison with A group, and the differences were statistically significant(P<0.01 or P<0.05).Conclusion High negative capacity is recommended in postoperative drainage of modified radical mastectomy,with a minimum of the equivalence of maximum daily drainage volume.If not,drainage in early postoperative duration will not be sufficient enough to insure complete adherence of skin flap,and postponed extubation and subcutaneous fluid accumulation would be expected.

Research progress in toxicology of molecular targeted anticancer drugs / 浙江大学学报·医学版

Drug therapy is essential for cancer treatment. The molecular targeted anticancer drugs develop rapidly in recent years, since the effectiveness of traditional chemotherapy is unsatisfactory and the adverse reactions are high. However, molecular targeted anticancer drugs would damage the function of heart, liver or lung, and may cause adverse effects such as hand-foot syndrome, which restrains their clinical application. Therefore, it is critical for pharmaceutical toxicology to study the toxicity, the related mechanisms and the preventive measures of molecular targeted anticancer drugs.

Therapeutic mechanism of bleomycin A5 on infancy hemangioma: an experimental study / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic mechanism of Bleomycin A5 on infancy hemangioma.</p><p><b>METHODS</b>After intralesional injection of Bleomycin A5 into the tumor of animal model of infancy hemangioma, the variation of tumor form was and the variation of tumor structure were observed using light microscope and electron microscope, the variation of tumor gene expression spectra was also tested by DNA microarray technique.</p><p><b>RESULTS</b>After treatment, the tumor gradually shrunk, hardened, disappeared one month later. The tumor lost appearance of infancy hemangioma and replaced by lamellar collagen fibers and cellular nucleus scattered in the fibers, and almost all cells were necrotic and dissolved. Under electron microscope, only large stretches of dissolved cell could be seen without intact cells and blood vessels, but apoptotic cells and bodies could also be found. The results of DNA microarray analysis showed that 9 genes associated with apoptosis (murine double minute 2, heat-labile enterotoxin B subunit, lymphotoxin B receptor, tumor necrosis factor ligand superfamily 7, tumor necrosis factor receptor superfamily 21, tumor necrosis factor receptor superfamily 1A, myeloid cell leukemia-1, caspase3), 13 genes associated with cell proliferation and cell cycle (cell division cycle27, cell division cycle37, CDC28 protein kinase 1B, cycling B1, cullin 2, cullin 3, cullin 4A, growth arrest and DNA damage-inducible 45A, meiotic recombination 11 homolog B, forkhead box M1, minichromosome maintenance 7, antigen identified by monoclonal antibody ki 67, proliferating cell nuclear antigen), and 11 genes associated with cellular stress and toxic reaction (glutathione peroxidase 1, metallothioneins, superoxide dismutase-1, heat shock protein A1A, heat shock protein A2, heat shock protein A4, heat shock protein A5, heat shock protein 9B, heat shock protein CA, macrophage migration inhibitory factor, plasminogen activator inhibitor)were up or down regulated more than 2 folds in tumors treated with Bleomycin A5 compared with controls.</p><p><b>CONCLUSIONS</b>The therapeutic effect of Bleomycin A5 on infancy hemangioma is the synthetic results of multiple factors. Bleomycin A5 could not only induce apoptosis and inhibit cell proliferation, but also depressed the ability of cell stress and toxic reaction.</p>

Effect of bleomycin A5 on the hemangioma-derived endothelial cell line XTPS-1 / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of bleomycin A5 on the hemangioma-derived endothelial cell line XTPS-1.</p><p><b>METHODS</b>Hemangioma-derived endothelial cell line XTPS-1 was cultured with different concentration of bleomycin A5 (1000, 100, 10, 1, 0 mg/L), and then the survival rate was measured by methyl thiazolyl terazolium (MTT), the variation of cell morphology was observed using inverted phase contrast microscope and electron microscope, the variation of cell cycle and apoptosis rate were measured using flow cytometry.</p><p><b>RESULTS</b>After 24 hours culture the cell survival rate was (92.96 ± 3.66)% and (99.86 ± 0.12)% in lower saturation group (10 and 1 mg/L), but (34.08 ± 3.11)% and (43.28 ± 2.88)% in higher saturation group (1000 and 100 mg/L). The difference between them was more significant (P < 0.01). Lower saturation of bleomycin A5 (10 and 1 mg/L)could induce apoptosis but had almost no cytotoxic effect. Higher saturation of bleomycin A5 (1000 and 100 mg/L) not only induced apoptosis, but also had strong cytotoxic effect, which was concentration dependent.</p><p><b>CONCLUSIONS</b>bleomycin A5 could induce apoptosis, inhibit cell proliferation and has direct cytotoxic effect.</p>

Establishment of human infancy hemangioma-derived endothelial cell line XPTS-1 and animal model of human infancy hemangioma / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To establish an immortalized human infancy hemangioma-derived endothelial cell line (HemEC) and animal model of human infancy hemangioma.</p><p><b>METHODS</b>Hemangioma-derived endothelial cells from specimen of human infancy hemangioma were cultured in vitro and monocloed, and then its growth curve was made, karyomorphism of chromosome analyzed, morphologic characteristics observe, factor VIII related antigen identified by immunohistochemical method.Vascular endothelial growth factor receptor 2 (VEGFR-2) was detected by flow cytometry. HemEC were inoculated subcutaneously in athymic mouse to establish animal model of infancy hemangioma. The animal model was observed closely and its pathological characteristic was also studied.</p><p><b>RESULTS</b>The cultural cells grew active, and immortalized spontaneously when they were subcultured on sixteenth generation. This cell line was cultivated for more than 70 times within one year and in good condition after freezing and resuscitating once and again, and had the morphologic character of HemEC. The cell population doubling time was 22 h. Factor VIII and VEGFR-2 were expressed positively. Karyo type analysis of the cell line showed abnormal diploid with the modal chromosomal number varying between diploid and triploid. The cell line was then named XPTS-1. The animal model of infancy hemangioma was successfully established and its character of histopathology was similar with that of infancy hemangioma.</p><p><b>CONCLUSIONS</b>The cell line of HemEC was successfully established and immortalized spontaneously, and had the morphologic and biological character of HemEC. The animal model of infancy hemangioma was successfully established and showed the character of histopathology similar with that of infancy hemangioma.</p>

Adherence test of Staphylococcus epidermidis on the surface of different material intraocular lenses / 中华实验眼科杂志

Background Postoperative endophthalmitis following intraocular lens(IOL)implantation is still one of the most feared complications of cataract surgery.Bacterial adhesion to IOLs during their insertion is a prominent etiological factor.The adhesion characteristics of bacteria to IOL are very vital for the prevention of endophthalmitis after cataract surgery.Objective The present study was to observe the in vitro adherence ability of Staphylococcus epidermidis to different intraocular lenses(IOLs)and compare the results in bacterial counting between scanning electron microscopy(SEM)photographs and quantitative cultures. Methods Five types of IOLs,including hydrophobic acrylic IOL,polymethylmethaerylate(PMMA)IOL,heparin-surface-modified(HSM) PMMA IOL,silicone(SI)IOL and hydrophilic acrylic IOL,were put into S.epidermidis(ATCC 12228)suspension for 1 hour.The bacterial adhesion numbers on the IOL surfaces were counted by quantitative cultures and scanning electron microscopy(SEM) photographs. Results Quantitative culture counting of viable adherent bacteria released by sonication showed that hydrophobic acrylic IOL and PMMA IOL were more likely for bacteria to attach.The number of bacteria on the five types of IOL surfaces showed significant differences(F=100.084,P=0.000).No significant differences were found in the number of bacteria between hydrophilic acrylic IOL and HSM-PMMA IOL (t=2.285,P=0.052)with the quantitative culture method.Direct counting of adherent bacteria in SEM photographs revealed that there were significant differences in bacterial adhesion numbers among difierent IOL material groups,with the numbers from high to low as follows:Hydrophobic IOL＞PMMA IOL＞SI IOL＞Hydrophilic IOL＞HSM-PMMA IOL(F=118.065,P=0.000).The counting method by SEM method was superior to that by quantitative cultures (t=5.019,P=0.000). Conclusion The bacterial adhesion ability varies upon the difference of IOL materials.Less bacterial adhesion is found on hydrophilic acrylic IOL and HSM-PMMA IOL,implying that the use of IOLs made from these two materials during surgery could diminish the incidence of postoperative endophthalmitis and intraocular inflammation associated with IOLs implantation.