Objective To observe the effects of Bushen Huoxue Prescription on the pathway related to necroptosis of nucleus pulposus cells in a model rabbit of intervertebral disc degeneration;To explore its mechanisms in delaying intervertebral disc degeneration.Methods A intervertebral disc degeneration rabbit model was established using the spinal instability method.Totally 40 model rabbits were randomly divided into model group,ibuprofen group and Bushen Huoxue Prescription low-,medium-and high-dosage groups.Additionally,a normal control group and a sham-operation group were set up,with 8 rabbits in each group.Each treatment groups received the corresponding drugs via gavage for two consecutive weeks.HE staining was used to observe morphology of nucleus pulposus tissue,transmission electron microscopy was used to observe ultrastructure in nucleus pulposus cells,immunohistochemical staining was used to assess the expressions of Aggrecan and Collagen Ⅱ in nucleus pulposus tissue,Western blot was used to detect the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein in nucleus pulposus tissue.Results Compared with the sham-operation group,the model group showed a significant decrease in nucleus pulposus cells,disordered cell arrangement,reduced extracellular matrix,interrupted cell membrane continuity under transmission electron microscopy,organelle swelling,nuclear membrane disruption,partial chromatin loss,and positive expression of Aggrecan and Collagen Ⅱ in nucleus pulposus tissue decreased(P<0.01),while the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein significantly increased(P<0.01).Compared with the model group,the treatment groups showed an increased number of nucleus pulposus cells with orderly arrangement and more extracellular matrix,the ultrastructural damage of the cell membrane,organelle and nucleus in nucleus pulposus cells was partially restored under transmission electron microscopy,the positive expressions of Aggrecan and Collagen Ⅱ significantly increased in Bushen Huoxue Prescription medium-and high-dosage groups and the ibuprofen group(P<0.05,P<0.01),while the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein significantly decreased(P<0.05,P<0.01).Conclusion Bushen Huoxue Prescription may delay intervertebral disc degeneration of the model rabbit by inhibiting the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein in nucleus pulposus cells,and promoting the generation of extracellular matrix components Aggrecan and Collagen Ⅱ.

Objective To observe the effects of Bushen Huoxue Prescription on pressure-induced necroptosis in human nucleus pulposus cells and the expressions of RIPK1/RIPK3/MLKL pathway;To explore its potential mechanism in delaying intervertebral disc degeneration.Methods Human primary nucleus pulposus cells were cultured in vitro,and a model of nucleus pulposus cell degeneration was established using continuous load pressure method.After modeling,the nucleus pulposus cells were divided into model group,Bushen Huoxue Prescription group and inhibitor group,blank serum,Bushen Huoxue Prescription containing serum and necroptotic apoptosis inhibitor(Nec-1)intervention were administered,respectively.Normal group nucleus pulposus cells were cultured routinely.AO/EB fluorescence dual staining method was used for detecting cell apoptosis,flow cytometry was used to detect the necroptosis rate of nucleus pulposus cells,Western blot was used to detect the protein expressions of p-receptor interacting protein kinase(RIPK)1,p-RIPK3 and p-mixed lineage kinase domain like protein(MLKL),RT-qPCR was used to detect the mRNA expressions of RIPK1,RIPK3 and MLKL.Results Compared with the normal group,the model group showed more red fluorescence under AO/EB staining of nucleus pulposus cells,which were round and condensed,the necroptosis rate of nucleus pulposus cells increased(P<0.05),the protein expressions of p-RIPK1,p-RIPK3 and p-MLKL increased(P<0.05,P<0.01),while there was no significant difference in RIPK1 mRNA expression(P>0.05),and RIPK3 and MLKL mRNA expression increased(P<0.01).Compared with the model group,Bushen Huoxue Prescription group and the inhibitor group had less red condensed chromatin in the nucleus pulposus cells,Bushen Huoxue Prescription group had a lower rate of necroptosis(P<0.05),while the inhibitor group showed a decreasing trend in necroptosis rate(P>0.05),the protein expressions of p-RIPK1,p-RIPK3 and p-MLKL decreased in Bushen Huoxue Prescription group and the inhibitor group(P<0.05,P<0.01),while there was no significant difference in RIPK1 mRNA expression(P>0.05),and RIPK3 and MLKL mRNA expressions decreased(P<0.01).Conclusion Bushen Huoxue Prescription can alleviate pressure-induced damage to nucleus pulposus cells and inhibit necroptosis,thereby slowing the progression of intervertebral disc degeneration.Its mechanism may be related to the RIPK1/RIPK3/MLKL pathway mediated necroptosis.

Objective To observe the effects of Bushen Huoxue Prescription on the pathway related to necroptosis of nucleus pulposus cells in a model rabbit of intervertebral disc degeneration;To explore its mechanisms in delaying intervertebral disc degeneration.Methods A intervertebral disc degeneration rabbit model was established using the spinal instability method.Totally 40 model rabbits were randomly divided into model group,ibuprofen group and Bushen Huoxue Prescription low-,medium-and high-dosage groups.Additionally,a normal control group and a sham-operation group were set up,with 8 rabbits in each group.Each treatment groups received the corresponding drugs via gavage for two consecutive weeks.HE staining was used to observe morphology of nucleus pulposus tissue,transmission electron microscopy was used to observe ultrastructure in nucleus pulposus cells,immunohistochemical staining was used to assess the expressions of Aggrecan and Collagen Ⅱ in nucleus pulposus tissue,Western blot was used to detect the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein in nucleus pulposus tissue.Results Compared with the sham-operation group,the model group showed a significant decrease in nucleus pulposus cells,disordered cell arrangement,reduced extracellular matrix,interrupted cell membrane continuity under transmission electron microscopy,organelle swelling,nuclear membrane disruption,partial chromatin loss,and positive expression of Aggrecan and Collagen Ⅱ in nucleus pulposus tissue decreased(P<0.01),while the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein significantly increased(P<0.01).Compared with the model group,the treatment groups showed an increased number of nucleus pulposus cells with orderly arrangement and more extracellular matrix,the ultrastructural damage of the cell membrane,organelle and nucleus in nucleus pulposus cells was partially restored under transmission electron microscopy,the positive expressions of Aggrecan and Collagen Ⅱ significantly increased in Bushen Huoxue Prescription medium-and high-dosage groups and the ibuprofen group(P<0.05,P<0.01),while the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein significantly decreased(P<0.05,P<0.01).Conclusion Bushen Huoxue Prescription may delay intervertebral disc degeneration of the model rabbit by inhibiting the expressions of p-RIPK1,p-RIPK3 and p-MLKL protein in nucleus pulposus cells,and promoting the generation of extracellular matrix components Aggrecan and Collagen Ⅱ.

Objective To observe the effects of Bushen Huoxue Prescription on pressure-induced necroptosis in human nucleus pulposus cells and the expressions of RIPK1/RIPK3/MLKL pathway;To explore its potential mechanism in delaying intervertebral disc degeneration.Methods Human primary nucleus pulposus cells were cultured in vitro,and a model of nucleus pulposus cell degeneration was established using continuous load pressure method.After modeling,the nucleus pulposus cells were divided into model group,Bushen Huoxue Prescription group and inhibitor group,blank serum,Bushen Huoxue Prescription containing serum and necroptotic apoptosis inhibitor(Nec-1)intervention were administered,respectively.Normal group nucleus pulposus cells were cultured routinely.AO/EB fluorescence dual staining method was used for detecting cell apoptosis,flow cytometry was used to detect the necroptosis rate of nucleus pulposus cells,Western blot was used to detect the protein expressions of p-receptor interacting protein kinase(RIPK)1,p-RIPK3 and p-mixed lineage kinase domain like protein(MLKL),RT-qPCR was used to detect the mRNA expressions of RIPK1,RIPK3 and MLKL.Results Compared with the normal group,the model group showed more red fluorescence under AO/EB staining of nucleus pulposus cells,which were round and condensed,the necroptosis rate of nucleus pulposus cells increased(P<0.05),the protein expressions of p-RIPK1,p-RIPK3 and p-MLKL increased(P<0.05,P<0.01),while there was no significant difference in RIPK1 mRNA expression(P>0.05),and RIPK3 and MLKL mRNA expression increased(P<0.01).Compared with the model group,Bushen Huoxue Prescription group and the inhibitor group had less red condensed chromatin in the nucleus pulposus cells,Bushen Huoxue Prescription group had a lower rate of necroptosis(P<0.05),while the inhibitor group showed a decreasing trend in necroptosis rate(P>0.05),the protein expressions of p-RIPK1,p-RIPK3 and p-MLKL decreased in Bushen Huoxue Prescription group and the inhibitor group(P<0.05,P<0.01),while there was no significant difference in RIPK1 mRNA expression(P>0.05),and RIPK3 and MLKL mRNA expressions decreased(P<0.01).Conclusion Bushen Huoxue Prescription can alleviate pressure-induced damage to nucleus pulposus cells and inhibit necroptosis,thereby slowing the progression of intervertebral disc degeneration.Its mechanism may be related to the RIPK1/RIPK3/MLKL pathway mediated necroptosis.

Objective: To study the correlation of antibody titer and immune balance after infection of mycoplasma pneumoniae in children with bronchial asthma. Methods: From October 2010 to August 2015, 108 children with bronchial asthma infected with mycoplasma pneumoniae were selected.There were 71 patients in the observation group, children with mycoplasma pneumoniae antibody titers >1∶160, and 37 cases were selected as control group, mycoplasma pneumonia antibody titer≤1∶160.The titer of mycoplasma antibody, serum interferon gamma(IFN-γ), interleukin-4(IL-4) and interleukin-17(IL-17) were measured. Results: The serum IFN-γ, IL-4 and IL-17 levels in the control group were (41.04±3.13)μg/L, (17.04±3.16)μg/L and (30.81±4.23)μg/L, respectively, which in the observation group were (33.26±3.17)μg/L, (24.15±5.09)μg/L and (41.29±5.09)μg/L, respectively.The serum IL-4 and IL-17 levels of the observation group were significantly higher than those of the control group(t=5.126, 3.429, all P<0.05). After infection of mycoplasma pneumoniae, the serum IFN-γ in the observation group decreased significantly(t=4.168, P<0.05). Conclusion The titer of antibody after mycoplasma pneumoniae infection in children with bronchial asthma is closely related to the immune balance.The higher the antibody titer, the more serious the immune imbalance.

Objective To investigate the effects of cataract surgery with transparent corneal incision in different directions,and discuss the application of dual channel visual quality analysis system in the evaluation of visual quality before and after cataract surgery.Methods Sixty-four patients (64 eyes) with cataract phacoemulsification combined with intraocuiar lens (IOL) implantation were enrolled in our hospital from January to May in 2016,and randomly divided into 2 groups:Group A with corneal incision in the temporal side of the eyes,group B with upper corneal incision.All the cases were examined with standard logarithmic optical acuity and dual-channel optical quality analysis system.The visual acuity,MTF cutoff,Strehl ratio (SR),objective scattering index (OSI) and contrast ratio were compared between the two groups.The OQAS values were 100％,20％ and 9％.Results (1) The visual acuity and corrected visual acuity of all patients increased gradually from postoperative 1 day to 1 week (P ＜0.05),and the visual acuity gradually stabilized after 1 week.The uncorrected visual acuity and corrected visual acuity at postoperative 1 day and 1 week in group A were significantly higher than those in group B at the same time point(P ＜ 0.05).There was no significant difference in uncorrected visual acuity and corrected visual acuity between the two groups at postoperative 1 month and 3 months (P ＞ 0.05).(2) At postoperative 1 day and 1 week,the OSI values of two groups were higher than those before operation,and all the time points were statistically significant (all P ＜ 0.05).The OSI values of group A at postoperative 1 day(2.81 ±0.89) and 1 week(1.98 ±0.73) were significantly lower than those of group B at 1 day(3.46 ± 1.17) and 1 week(2.61 ± 1.20) (all P ＜ 0.05).There was no significant difference in OSI between postoperative 1 month and 3 months (all P ＞ 0.05).The OV9％ values of the two groups were increased at postoperative 1 day,1 week,1 month and 3 months,and all the time points were statistically significant(all P＜0.05).The value of OV9％ in group A at postoperative 1 day(0.29 ±0.09),1 week(0.33 ±0.12),1 month (0.37 ± 0.14) and 3 months (0.42 ± 0.17) were significantly higher than those in group B at 1 day (0.24 ± 0.10),1 week (0.27 ± 0.09),1 month (0.31 ± 0.09) and 3 months (0.34 ± 0.14),which was statistically significant (all P ＜ 0.05).The MTF,SR,OV100％ and OV20％ values of all the patients were not statistically significant (all P ＞ 0.05).Conclusion In the operation of cataract with the temporal and transparent corneal incision,the postoperative intraocular scattering is small,early visual recovery is fast and the visual quality is high.The long-term effect needs further study.Dual channel visual quality analysis system can further quantify the visual quality parameters,has certain advantage in visual quality assessment.

Objective To know the nursing students' willingness engaging in geriatric nursing and analyze affecting factors so as to provide references for nurturing professionals for caring for the aged.Methods 231 nursing students were surveyed.The questionnaire included general information,life experience with the elderly,willingness engaging in geriatric nursing,attitude towards the elderly and cognition of aging.The investigation results underwent analysis.Results Single factor analysis showed that whether ever living with the elderly (x2=6.027),time living with the elderly and relationship with the elderly (x2=4.928) influeced the nursing students' willingness engaging in geriatric nursing.Multiple correspondence analysis indicated that nursing students living with elderly more than one year and having good relationship with the elderly had positive willingness.Conclusions Geriatric nursing curriculum should be optimized and more opportunities should be created for nursing students to be in touch with the elderly.At the same time,we must pay attention to occupation guide and moral education about how to respect and care for the elderly.

Objective To know the effects of living with the elderly on nursing students' willingness engaging in aged nursing and attitude towards the elderly so as to provide references for nurturing professionals for caring for the aged.Methods 231 nursing students were surveyed.The questionnaire included four parts,general information,life experience with the elderly,willingness engaging in aged nursing,attitude towards the elderly and cognition of aging.Results 45.9 percent of the nursing students were willing to engage in aged nursing.Living with elderly and relationship with the elderly affected their willingness.Conclusions Creating more opportunities for nursing students in touch with the elderly can help nursing students to have a good impression of the elderly and stimulate them to choose aged nursing.

OBJECTIVE: To observe growth inhibition effect of perlecan anti-sense cDNA (pAP) on human laryngeal carcinoma xnografted in nude mice. To vertify its antitumor effect and mechanism in vivo, and it may be useful as a biomarker in carcinoma of larynx cancer. METHOD: Created the model of human laryngeal carcinoma xnograft in nude mice. To observe growth of those xnografts in nude mice and draw growth curve of xnografted. The expression of perlecan mRNA and portein in xnografts were examined by RT-PCR and immunohistochemistry. RESULT: Volume of xnografts in the group transfected by the plasmids of pAP were significant small as compared with other two groups made by the wild type cells and phpApr-neol cells (P < 0.05). It was showed that the expression of perlecan mRNA and protein were significantly reduced in the tumor of pAP transfected Hep-2 cells as compared with the tumors transfected by the wild type cells and phβApr-neol cells (P < 0.01). CONCLUSION These data raise the possibility that pAP many play key roles in the growth of those xnografts in nude mice.
Animals ; DNA, Antisense ; therapeutic use ; DNA, Complementary ; Heparan Sulfate Proteoglycans ; genetics ; Heterografts ; Humans ; Laryngeal Neoplasms ; pathology ; therapy ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Plasmids ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Objective To evaluate the outcome of combination of intensive preconditioning regimen allo-HSCT with imatinib for treatment of Ph chromosome positive acute lymphocyte leukemia (ALL). Methods Between 2009 and 2010, 8 patients diagnosed as Ph+ ALL received allo-HSCT from HLA identical sibling during complete remission. Imatinib was added into the therapies of 5 patients.Seven patients received the intensive preconditioning regimen based on BuCy2, one patient received the regimen of TBI-Cy. A median of 6. 02 × 108/kg mononuclear cells and 3. 14 × 106/kg CD34+ cells were transfused. GVHD prophylaxis included cyclosporine A and methotrexate. Results All patients were well tolerant to the regimen without serious regimen-related toxicity. The median time of ANC≥0. 5 × 109/L was 15. 5 days, and that of PLT≥20 × 109/L was 19 days. Thirty days after allo-HSCT, all patients got donor engraftment successfully. Among 8 cases, 4 cases presented acute GVHD, 2 developed degree Ⅰ , one developed degree Ⅱ , and one developed degree Ⅳ. Seven patients were alive 100 days after allo-HSCT, 3 of whom presented chronic GVHD. At the end of following-up period, 6 patients were alive, among them, 3 patients were alive without relapse; 3 patients relapsed; Two patients died, one from acute GVHD, and one from leukemia relapse. Conclusion Combined intensive preconditioning regimen allo-HSCT with Imatinib was an effective treatment for Ph+ ALL, but the effect of anti-chronic GVHD of imatinib should arouse certain attention.