Objective To explore the effect of nicotine on the degeneration of dopaminergic neurons in PD rats. Methods Forty-five SD rats were randomly divided into three groups: PBS group (CON), normal saline + LPS ( NS ) group and nicotine +LPS ( NIC ) group. On 24 hours after LPS or PBS injection, inducible nitric oxide synthase ( iNOS ) protein expression was examined by immunoblotting. ON 14d after LPS or PBS injection, the numbers of tyrosine hydroxylase ( TH ) positive neurons and morphological changes of OX-42 positive cells in the substantia nigra(SN) were observed by immunohistochemistry. TH mRNA and TH protein expressions were examined by RT-PCR or immunoblotting. Results Compared with CON group, iNOS protein increased markedly 24 hours after LPS injection in NS group. TH positive neurons, TH mRNA and TH protein in the SN decreased remarkably 14 days after nigrainjiection. Most of microlglial showed big cell body with thicker and shorter processes. However, nicotine reversed the above changes, Compared with NS group, TH positive neurons, TH mRNA and TH protein in the SN increased remarkably in NIC. Besides, most microglia showed small cell body with slim and long processes. Conclusion Nicotine could prevent LPS-induced degeneration of DA neurons, probably because of that the pretreatment with nicotine blocks the activation of microglia and the expression iNOS protein.

Objective To investigate the intervention effect of integrated care including exercise therapy,example learning and psychological support to ease the hematopoietic stem cell transplantation (HSCT) related complications.Methods 40 cases were divided into the intervention group and the control group according to their hospital number,there were 20 cases in each group.The intervention group received integrated care and control group adopted traditional methods.HSCT symptoms scale was used to evaluate the intervention effects in 6 time points (the day before pretreatment,the day before HSCT,the 7th day,14th day,21th day and three months after HSCT).Results There were no statistical differences between the two groups in the score of HSCT scales on the day before pretreatment.The intervention group was statistically lower than the control group in the score of HSCT scales on the day before HSCT,the 7th day,14th day,21th day and three months after HSCT,P＜0.05.Conclusion The integrated care can reduce HSCT complications and worthybe popularizing.

Objective To study whether the positive psychological capital intervention could improve the level of psychological capital and the will of retention of clinical nurses. Methods A randomized controlled trials was conducted. The control group did not take any psychological intervention;Intervention group was given psychological capital intervention, which was conducted for 7 weeks (once a week, 1-2 hours per time).It used Nurses′ Psychological Capital Questionnaire and Nurses′ Retention Intention Questionnaire to evaluate the participants before and after intervention. Results There were no statistical differences between intervention group and control group in the score of Nurses′Psychological Capital Questionnaire (84.74 ± 10.04 vs. 86.16 ± 8.57,P > 0.05 ) and Nurses′ Retention Intention Questionnaire ( 19.21±3.47 vs. 18.54±3.41,P>0.05 ) on the day before intervention. The score of Nurses′Psychological Capital Questionnaire of intervention group was 93.89 ± 7.10 on the intervention day and 93.60 ± 6.88 three months later while the control group scored 84.41 ± 6.58 and 83.92 ± 9.31.The intervention group was statistically lower than the control group (t=6.156, 5.155, P<0.05) .And the score of Nurses Retention Intention Questionnaire was 22.42±2.66 on the intervention day and 22.24±2.61 three months later while the control group scored 18.61 ± 3.08 and 19.21 ± 3.47. The intervention group was statistically lower than the control group (t=5.866, 4.292, P<0.05). Conclusions Positive psychological capital intervention can improve the the will of retention of clinical nurses.

Objective To observe the effect of nicotine(NIC) on the activation and resultant death of microglia induced by LPS. Methods The animal model that exposed to chronic nicotine treatment was established and LPS was injected intraperitoneally to induce the activation of microglia. Furthermore, the CD11b-positive microglia in cerebral cortex, hippocampal and substantia ngra were observed through immunohistochemical staining. BV2 cells(Microglial cell line of mouse) were subcultured, simultaneously the following kits were used including CCK-8 kit assay for cell activity, Nitric oxide assay kit assay for NO release, RT-PCR assay for the iNOS,TNF-α,IL-1β,IL-6,COX-2,IRF-1,Caspase-11 mRNA expression, Western blotting assay for the protein expression of P-I-κB and Caspase-3. Results Nicotine suppressed the CD11b-positive microglia expression in cerebral cortex,hippocampal and substantia ngra induced by LPS;Nicotine inhibited the activation-induced cell death (AICD), attenuated NO release, reduced iNOS,TNF-α,IL-1β,IL-6,COX-2,IRF-1,Caspase-11 mRNA expression, decreased the protein expression including P-I-κB and Caspase-3 of BV2 cells. Conclusion Nicotine pretreatment can suppress the activation and resultant death of microglial cells induced by LPS, which suggests that nicotine may play a neuroprotective role on inflammatory reaction of brain.;

Objective To evaluate the effectives of high-frequency oscillatory ventilation (HFOV)in the treatment of severe NRDS.Methods The severe NRDS who failed to response to conventional mechanical ventilation (CMV) were changed to HFOV treatment.Ventilator parameters and blood gas analysis were compared before and after treatment.Results Among thirty patients,19 cases were significantly improved and 11 cases were failed,the success rate was 63％.Nineteen patients with HFOV,the main ventilator parameters and blood gas results such as FiO2 ( 0 h:0.87 ± 0.09 vs 3 h:0.67 ± 0.10,6 h:0.54 ±0.08,12h:0.44 ±0.08),MAP[0 h:(14.47 ±1.17) mbar vs 3 h:(13.79 ± 1.03) mbar,6 h:(13.26 ±1.05) mbar,12 h:( 12.37 ±0.83) mbar],f[0 h:(7.26 ±0.56) Hz vs 3 h:(7.89 ±0.57) Hz,6 h:(8.42±0.51 ) Hz,12 h:(9.26 ±0.45) Hz] and PaCO2 [0 h:(9.69 ±0.86) kPa vs 3 h:(6.00 ±0.55) kPa,6 h:(5.62 ± 1.03)kPa,12 h:(5.47 ±0.55)kPa] were decreased while pH(0 h:7.21 ±0.06 vs 3 h:7.35 ±0.07,6 h:7.39 ±0.07,12 h:7.40 ±0.06) and PaO2[0 h:(5.46 ±0.82)kPa vs 3 h:(7.92 ± 1.13)kPa,6 h:(9.25 ± 0.99)kPa,12 h:(9.20 ± 0.96)kPa] were significantly increased compared with before treatment ( P ＜ 0.05).Conclusions The patients of severe NRDS who failed to response to CMV were treated with HFOV,some patients received a significant effect with HFOV treatment.

Objective To compare the efficacy and safety of excimer laser 308 nm phototherapy alone and the combination of excimer laser 308 nm and topical application of vitamine D3 alanogue tacalcitol in the treatment of vitiligo. Methods Seventy-eight patients with vitiligo were enrolled in the single-blind clinical trial, treated with excimer laser 308 nm. The lesions were devided into two groups: patients in the experimental group were instructed to use tacalcitol ointment and the control group were applied with placebo ointment. The lesions were evaluated once per month and photos taken for analyses of clinical effects. Results The results in different locations were compared, the effective rates of the experimental group in cephalofacial site, trunk and limbs were 93.51%, 84.16 % and 42.35 %, respectively. The effective rates of control group in opposite and adjacent sites were 90.9 %, 77.45 % and 34.15 %, respectively (P ＜ 0.05). The comparison of results in different types of lesions indicated that the effective rate of the experimental group in vitiligo vulgaris and segmental vitiligo were 73.81% and 84.00 %, respectively. The effective rate of control group in vulgaris and segmental vitiligo were 86.8 %, 77.45 % and 34.15 %, respectively (P ＜0.05 ). The comparison of results in radiation times and doses of phototherapy showed that the radiation time and dose on the time of initial pigment regeneration were (16. 15 ± 3.22)times and (4.40 ± 5.03)J/cm2 in the experimental group, while ( 18.56 ± 3.50) times and ( 6.60 ± 1.01 ) J/cm2 ( P ＜ 0.05 ) in the control group, the time and dose on the time of apparent effect were ( 20. 36 ± 1.50 ) times and ( 7.50 ± 3.54 ) J/cm2 in the experimental group, and (21.68 ± 2.40) times and( 8.80 ± 9.24)J/cm2 (P ＜ 0.05 ) in the control group. Conclusion Application of tacalcitol ointment in combination with twice-weekly 308 nm excimer laser light phototherapy is an effective alternative treatment for patients with generalized vitiligo.

This study was aimed to explore the curative effect of Zhuo-Du-Qing (ZDQ) particles on diabetic cystopa-thy (DCP) based on the theory of turbid toxin. A total of 100 DCP patients were randomly divided into the control group with 47 cases and the treatment group with 53 cases. Mecobalamin tablets was given in the control group. And ZDQ particles were given to the treatment group. The treatment course was 3 months. The observation was made on the bladder residual urine (BRU), clinical manifestation integral, hemodynamic indexes, HOMA-IR and HbA1c before and after the treatment. The results showed that compared with pretreatment, the BRU, clinical manifestation integral, hemodynamic indexes, and HOMA-IR were significantly reduced (P < 0.01). Compared with the control group, the BRU, clinical manifestation integral, hemodynamic indexes, and HOMA-IR were also significantly reduced (P <0.01). The effective rate in the treatment group was 81.1%, which was significantly higher than 59.6% in the control group (P< 0.05). There were no obvious changes on the blood routine examination, urine routine examination, stool routine examination, liver function, renal function, electrocardiogram and so on before and after treatment. It was con-cluded that the treatment of DCP with ZDQ particles was safe and effective. To remove toxin and descend turbid may be another effective treatment method for DCP. The occurrence and development of DCP were closely related to the turbid toxin.

Pulldown assay is an in vitro method for studies of protein-protein interactions, in which tagged proteins are usually expressed as the bait to enrich other proteins that could bind to them. In this technology, the GST tag is broadest used for its modest size and hydrophilic property. In most cases, the GST tag could increase the hydrophility of the fusion protein and help to avoid the formation of inclusion bodies. However, in the other few cases, the target protein may be strongly hydrophobic or have complicated structures that were hard to fold and assemble in correct conformations without champerons, and even the existence of GST tag could not make them soluble. These proteins were always expressed as inclusion bodies and had no functions. LMO2 was a small molecular weight and insoluble protein, in this study, GST system and MBP system were used to express GST-LMO2 and MBP-LMO2 fusion proteins, respectively. We found that GST-LMO2 fusion protein was expressed as inclusion bodies whereas MBP-LMO2 fusion protein was expressed in soluble form. Moreover, the production rate of MBP-LMO2 was also much higher than GST-LMO2. Then MBP-LMO2 fusion proteins and renatured GST-LMO2 fusion proteins were used as bait in pulldown assay to study the interaction between LMO2 and endogenous GATA1 in K562 cells. Western blot analyses showed that both of these proteins could bind to endogenous GATA1 in K562 cells, but recovered GATA1 protein by MBP-LMO2 fusion protein was much more than GST-LMO2 fusion protein. These results suggest that using of MBP system is a helpful attempt in the case of studying small molecular weight, strong hydrophobic proteins.
Adaptor Proteins, Signal Transducing ; Carrier Proteins ; chemistry ; Chemical Precipitation ; DNA-Binding Proteins ; chemistry ; GATA1 Transcription Factor ; chemistry ; Genetic Vectors ; Glutathione Transferase ; chemistry ; Humans ; K562 Cells ; LIM Domain Proteins ; Maltose-Binding Proteins ; Metalloproteins ; chemistry ; Protein Binding ; Protein Interaction Domains and Motifs ; Protein Renaturation ; Proto-Oncogene Proteins ; chemistry ; Recombinant Fusion Proteins ; genetics ; metabolism

Bone morphogenetic protein 2 (BMP2), which belongs to the transforming growth factor-beta (TGF-beta) superfamily, is a multifunctional molecule with distinct abilities to induce bone formation. BMP2 has been identified to have eminent pharmaceutical importance for clinical application. We previously constructed stable cell line in Chinese hamster ovary cells (CHO) that highly expressed recombinant human BMP2 (rhBMP2). For large-scale production of the recombinant protein used in clinical application, it is critical to have both high expression and stability of the protein. In the present study, the stability of the cell line (rCHO(hBMP2)-C8) with the highest expression, as well as the stability of rhBMP2 protein were investigated systematically. We cultured the rCHO (hBMP2)-C8 cell line in the presence or absence of MTX for two months, the cell growth and rhBMP2 production characteristics were examined during the culture; we found the duration that the rCHO(hBMP2)-C8 cell line could secret rhBMP2 continually into the serum-free medium. Moreover, we detected the temperature sensitivity of rhBMP2 in culture medium. This study will contribute to our understanding for further producing rhBMP2 by large-scale culture technology.
Animals ; Bone Morphogenetic Protein 2 ; biosynthesis ; genetics ; CHO Cells ; Cell Culture Techniques ; methods ; Cricetinae ; Cricetulus ; Culture Media ; Genetic Vectors ; genetics ; Humans ; Recombinant Proteins ; biosynthesis ; genetics

Objective:To discuss the quantitative evaluation of left ventricular systolic function in patients with light-chain amyloidosis (AL) by pressure-strain loop (PSL).Methods:Forty-six patients with clinically diagnosed as AL in the Affiliated Tumor Hospital of Zhengzhou University from January 2018 to December 2020 were selected as case group, and they were divided into 2 groups according to whether the thickness of interventricular septum and posterior wall at end-diastole was >12 mm; ①cardiac amyloidosis (CA) group (21 cases, the thickness>12mm); ②non cardiac amyloidosis (NCA) group (25 cases, the thickness≤12 mm). Twenty five healthy volunteers were selected as control group at the same time. Routine echocardiography was performed in all subjects.Two-dimensional dynamic images of the left ventricular apical two-chamber, three-chamber, and four-chamber views were collected for three consecutive cardiac cycles using two-dimensional speckle-tracking. A tracing analysis was conducted and blood pressure was entered on the off-line Echo PAC 203 software, and the left ventricular global longitudinal strain (GLS), peak strain time dispersion(PSD), global myocardial work index (GWI), global constructive work (GCW), global wasted work (GWW) and global work efficiency (GWE) were obtained. The differences of GLS, PSD and myocardial work (GWI, GCW, GWW, GWE) parameters were compared between groups, and Pearson correlation was used to analyze the correlations.Results:①Routine echocardiography: Compared with the control group and NCA group, inter-ventricular septum thickness (IVST), left ventricle posterior wall thickness (LVPWT), left ventricular mass index (LVMI), left atrial diameter (LAD), and E/e′ in CA group were increased, while left ventricular ejection fraction (LVEF) was decreased; ②Compared with the control group, GLS, GWI, and GCW in NCA group were decreased, while there were no statistically differences in GWE, PSD, and GWW between the two groups (all P>0.05); Compared with the control group and NCA group, GLS, GWI, GCW, and GWE were obviously decreased, while PSD and GWW were obviously increased in CA group.③ Correlation analysis showed that: the absolute value of GLS was positively correlated with GWI, GCW and GWE ( r=0.654, 0.695, 0.788; all P<0.001), and negatively correlated with GWW, and PSD ( r=-0.710, -0.625; all P<0.001). Besides, PSD had negative correlation with GWI, GCW and GWE ( r=-0.754, -0.653, -0.702; all P<0.001), and positive correlation with GWW ( r=0.676, P<0.001). Conclusions:PSL can quantitatively evaluate the left ventricular systolic function of AL patients, while the myocardial work parameters are conducive to evaluate the degree of cardiac involvement in the course of AL patients.