Activation of D2-like dopamine receptors improves the neuronal network and cognitive function of PPT1KI mice.

Palmitoyl-protein thioesterase 1 (PPT1) is a lysosomal depalmitoylation enzyme that mediates protein posttranslational modifications. Loss-of-function mutation of PPT1 causes a failure of the lysosomal degradation of palmitoylated proteins and results in a congenital disease characterized by progressive neuronal degeneration referred to as infantile neuronal ceroid lipofuscinosis (INCL). A mouse knock-in model of PPT1 (PPT1-KI) was established by introducing the R151X mutation into exon 5 of the PPT1 gene, which exhibited INCL-like pathological lesions. We previously reported that hippocampal γ oscillations were impaired in PPT1 mice. Hippocampal γ oscillations can be enhanced by selective activation of the dopamine D4 receptor (DR4), a dopamine D2-like receptor. In this study, we investigated the changes in DR expression and the effects of dopamine and various DR agonists on neural network activity, cognition and motor function in PPT1KI mice. Cognition and motor defects were evaluated via Y-maze, novel object recognition and rotarod tests. Extracellular field potentials were elicited in hippocampal slices, and neuronal network oscillations in the gamma frequency band (γ oscillations) were induced by perfusion with kainic acid (200 nM). PPT1KI mice displayed progressive impairments in γ oscillations and hippocampus-related memory, as well as abnormal expression profiles of dopamine receptors with preserved expression of DR1 and 3, increased membrane expression of DR4 and decreased DR2 levels. The immunocytochemistry analysis revealed the colocalization of PPT1 with DR4 or DR2 in the soma and large dendrites of both WT and PPT1KI mice. Immunoprecipitation confirmed the interaction between PPT1 and DR4 or DR2. The impaired γ oscillations and cognitive functions were largely restored by the application of exogenous dopamine, the selective DR2 agonist quinpirole or the DR4 agonist A412997. Furthermore, the administration of A412997 (0.5 mg/kg, i.p.) significantly upregulated the activity of CaMKII in the hippocampus of 5-month-old PPT1KI mice. Collectively, these results suggest that the activation of D2-like dopamine receptors improves cognition and network activity in PPT1KI mice and that specific DR subunits may be potential targets for the intervention of neurodegenerative disorders, such as INCL.

AQP4-A25Q Point Mutation in Mice Depolymerizes Orthogonal Arrays of Particles and Decreases Polarized Expression of AQP4 Protein in Astrocytic Endfeet at the Blood-Brain Barrier.

Aquaporin-4 (AQP4) is characterized by the formation of orthogonal arrays of particles (OAPs) comprising its M1 and M23 isoforms in the plasma membrane. However, the biological importance of OAP formation is obscure. Here, we developed an OAP depolymerization male mouse model by transgenic knock-in of an AQP4-A25Q mutation. Analyses of the mutant brain tissue using blue native polyacrylamide gel electrophoresis, super-resolution imaging, and immunogold electron microscopy revealed remarkably reduced OAP structures and glial endfeet localization of the AQP4-A25Q mutant protein without effects on its overall mRNA and protein expression. AQP4A25Q/A25Q mice showed better survival and neurologic deficit scores when cerebral edema was induced by water intoxication or middle cerebral artery occlusion/reperfusion. The brain water content and swelling of pericapillary astrocytic endfeet processes in AQP4A25Q/A25Q mice were significantly reduced, functionally supporting decreased AQP4 protein expression at the blood-brain barrier. The infarct volume and neuronal damage were also reduced in AQP4A25Q/A25Q mice in the middle cerebral artery occlusion/reperfusion model. Astrocyte activation in the brain was alleviated in AQP4A25Q/A25Q mice, which may be associated with decreased cell swelling. We conclude that the OAP structure of AQP4 plays a key role in its polarized expression in astrocytic endfeet processes at the blood-brain barrier. Therefore, our study provided new insights into intervention of cerebral cellular edema caused by stroke and traumatic brain injury through regulating AQP4 OAP formation.SIGNIFICANCE STATEMENT Aquaporin-4 (AQP4) is characterized by orthogonal arrays of particles (OAPs) comprising the M1 and M23 isoforms in the membrane. Here, an OAP depolymerization male mouse model induced by AQP4-A25Q mutation was first established, and the functions of OAP depolymerization in cerebral edema have been studied. The results revealed that AQP4 lost its OAP structure without affecting AQP4 mRNA and protein levels in AQP4-A25Q mice. AQP4-A25Q mutation mice has neuroprotective effects on cerebral edema induced by water intoxication and middle cerebral artery occlusion/reperfusion through relieving the activation of astrocytes and suppressed microglia-mediated neuroinflammation. We concluded that the OAP structure of AQP4 plays a key role in its polarized expression in astrocytic endfeet processes at the blood-brain barrier. Therefore, our study provided new insights into intervention of cerebral cellular edema caused by stroke and traumatic brain injury through regulating AQP4 OAP formation.

Iodine-Mediated Domino Cyclization for One-Pot Synthesis of Indolizine-Fused Chromones via Metal-Free sp3 C-H Functionalization.

An efficient method for the synthesis of new indolizine-fused chromones has been accomplished from ethyl (E)-3-(2-acetylphenoxy)acrylates and pyridines in a "one-pot" manner. Facile operation in open-air, metal-free, and mild conditions renders this protocol particularly practical and attractive. Moreover, this method can simultaneously construct two molecular fragments of chromone and indolizine. Scale-up experiment and the construction of natural products further prove the practicability of this strategy.

[Clinical observation of pre-suture ligation and suture knot positioning in single-operator circumcision with the stapler for children].

OBJECTIVE: To observe the clinical effects of pre-suture ligation and suture knot positioning in single-operator circumcision with the stapler. METHODS: Totally 120 six to fourteen years old children with phimosis or redundant prepuce were equally and randomly assigned to receive traditional single-operator circumcision with the stapler (group 1), single-operator circumcision with double suture knots for positioning the cutting plane with the stapler (group 2), or pre-suture ligation plus single-operator suture knot positioning circumcision with the stapler (group 3). We recorded and comparatively analyzed the operation time, intraoperative blood loss and hematoma, the number of residual suture knots, the patients' satisfaction with foreskin reservation, sutured frenulum and incision aesthetics, and the rates of surgical conversion, severe postoperative dysuria, severe wound exudation and perioperative anxiety of the parents and surgeon. RESULTS: Statistically significant differences were observed among the three groups in the operation time, intraoperative blood loss, incidence of intraoperative hematoma, number of residual suture knots, the patients' satisfaction with foreskin reservation, sutured frenulum and incision aesthetics, and the rates of surgical conversion, severe postoperative dysuria, severe wound exudation and perioperative anxiety of the parents and surgeon (P < 0.05). Compared with group 1, group 3 showed obviously less operation time and intraoperative blood loss and lower incidence of intraoperative hematoma, number of residual suture knots and rates of surgical conversion and perioperative anxiety of the parents and surgeons, and higher rates of patients' satisfaction with foreskin reservation, sutured frenulum and incision aesthetics. The intraoperative blood loss, incidence of intraoperative hematoma, and the rates of severe postoperative dysuria, severe wound exudation and perioperative anxiety of the parents and surgeon were lower in group 3 than in group 2. CONCLUSION: Pre-suture ligation plus single-operator suture knot positioning circumcision with the stapler helps reduce the operation time, intraoperative blood loss, incidence of intraoperative hematoma, number of residual suture knots and rates of surgical conversion and perioperative anxiety of the parents and surgeon, and increase the patients' satisfaction with foreskin reservation, sutured frenulum and incision aesthetics.

[Single- versus double-suture knot for positioning the cutting plane in circumcision with a stapler].

OBJECTIVE: To observe the clinical effect of single-suture versus that of double-suture knot in positioning the cutting plane in circumcision with a stapler. METHODS: We randomly assigned 120 patients with redundant prepuce or phimosis into three groups of an equal number to receive traditional circumcision without suture knot (group 1), circumcision with single-suture knot (group 2), and circumcision with double-suture knot (group 3) for positioning of the cutting plane. We recorded and compared the operation time, intraoperative blood loss, the rates of frenulum sewing, non-frenulum sewing, poor frenulum sewing and surgical conversion, intraoperative anxiety of the doctors, postoperative ecchymosis, and satisfaction with the retained ventral and dorsal prepuce and postoperative penile appearance among the three groups. RESULTS: There were statistically significant differences among the three groups in the surgery time, intraoperative blood loss, the rates of frenulum sewing, non-frenulum sewing, poor frenulum sewing and surgical conversion, intraoperative anxiety of the doctors, and satisfaction with the retained ventral and dorsal prepuce and postoperative penile appearance, (P < 0.05), but not in postoperative ecchymosis (P = 0.849). The rate of satisfaction with the retained dorsal prepuce was remarkably higher in group 3 than in group 2 (P = 0.003), and the intraoperative anxiety rate of the doctors was lower in the former than in the latter group (P = 0.003). CONCLUSIONS: Both single- and double-suture knots for positioning the cutting plane in circumcision with a stapler can help reduce the operation time, intraoperative blood loss, the rates of frenulum sewing, non-frenulum sewing, poor frenulum sewing and surgical conversion, intraoperative anxiety of the doctors, and satisfaction with the retained ventral and dorsal prepuce and postoperative penile appearance, and double-suture knot positioning has an even higher application value in decreasing the intraoperative anxiety of the doctors and increasing the satisfaction with the retained dorsal prepuce.

[Correlation between the phenotypic domains of the UPOINT system and sexual dysfunction in men with chronic prostatitis/chronic pelvic pain syndrome].

OBJECTIVE: To investigate the correlation of the six phenotypic domains of the UPOINT (urinary, psychosocial, organ-specific, infection, neurologic/systemic, and tenderness) system with premature ejaculation (PE) and ED in male patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). METHODS: We collected the clinical data on 453 cases of CP/CPPS from December 2016 to December 2017, including the general information, CP symptoms, sexual life history, intravaginal ejaculatory latency time (IELT), NIH-CPSI, and IIEF-5 scores. We classified the patients according to the UPOINT system. RESULTS: The CP/CPPS patients were aged 32.75 ± 6.85 years, of whom 45.03% (204/453) were diagnosed with ED and 43.27% (196/453) with PE. The positive rates of the six phenotypic domains of the UPOINT system were 68.78% (U), 60.21% (P), 77.45% (O), 20.34% (I), 46.83% (N), and 65.12% (T), respectively. Multivariate logistic regression analysis showed psychosocial (P) abnormality to be an independent risk factor for PE (OR = 4.55, 95% CI: 2.75－8.06, P < 0.05) and ED (OR = 3.35, 95% CI: 2.02－6.25, P < 0.05). CONCLUSIONS: The psychosocial (P) factor in the UPOINT system plays an important role in the pathogenesis of PE and ED in patients with CP/CPPS.

[Study on liver protection and hepatotoxicity of saikosaponin a based on zebrafish model].

Bupleuri Radix has both liver protection and hepatotoxicity. Saponins are the main pharmacodynamic and toxic components of Bupleuri Radix. Based on zebrafish physical model and the model of alcoholic fatty liver( AFL) pathology,the liver toxic and protective effect of saikosaponin a( SSa) were assessed. The results indicated that 1. 77 µmol·L-1 SSa showed protective effect to AFL zebrafish. 5. 30 µmol·L-1 SSa was hepatotoxic to healthy zebrafish,but it showed protective effect to AFL zebrafish. 5. 62 µmol·L-1 SSa was hepatotoxic to healthy and AFL zebrafish. This study is benefit for clinical safety of saikosaponin a.

Parthenolide attenuated bleomycin-induced pulmonary fibrosis via the NF-κB/Snail signaling pathway.

BACKGROUND: Parthenolide (PTL) is a natural molecule isolated from Tanacetum parthenium that exhibits excellent anti-inflammatory and antitumor activities. Pulmonary fibrosis (PF), especially idiopathic pulmonary fibrosis (IPF), is a chronic lung disease that lacks a proven effective therapy. The present study evaluated the therapeutic effect of PTL on PF. METHODS: Serum-starved primary lung fibroblasts and HFL1 cells were treated with different doses of PTL, and cell viability and the migration rate were measured. Western blot analysis and a dual-luciferase assay were used to analyze the epithelial-mesenchymal transition (EMT)-related transcription factors influenced by PTL treatment in A549 cells and primary lung epithelial cells. Mice with bleomycin (BLM)-induced pulmonary fibrosis were treated with different doses of intragastric PTL, and pathological changes were evaluated using Hematoxylin-eosin (H&E) staining and immunohistochemical analysis. RESULTS: Our results demonstrated that PTL reduced the cell viability and migration rate of lung fibroblasts and inhibited the expression of EMT-related transcription factors in lung epithelial cells. In vivo studies demonstrated that PTL attenuated BLM-induced pulmonary fibrosis and improved the body weight and pathological changes of BLM-treated mice. We further demonstrated that PTL attenuated BLM-induced PF primarily via inhibition of the NF-κB/Snail signaling pathway. CONCLUSION: These findings suggest that PTL inhibits EMT and attenuates BLM-induced PF via the NF-κB/Snail signaling pathway. PTL is a worthwhile candidate compound for pulmonary fibrosis therapy.

[Correlation between premature ejaculation diagnostic tool and International Index of Erectile Function-15 in different types of premature ejaculation].

OBJECTIVE: To investigate the correlation between the premature ejaculation diagnostic tool (PEDT) and International Index of Erectile Function-15 (IIEF-15) in different types of premature ejaculation (PE). METHODS: We performed a cross-section survey among 352 PE patients received in the andrology clinic from December 2014 to December 2015 and 104 healthy men from the health examination center using basic demographic information (as on age, height, weight, education status, occupation, income, etc.), PEDT results, and IIEF-15 scores of the subjects. RESULTS: The PE patients had remarkably higher PEDT and lower IIEF-15 scores than the healthy men (P<0.01). The PEDT score of the PE patients was negatively correlated with their total IIEF-15 score as well as with the scores in the domains of erectile function, sexual intercourse satisfaction, and overall satisfaction after adjusted for age (P<0.01). The patients with acquired PE (APE) showed a lower IIEF-15 score than those with lifelong PE (LPE) (P<0.01). The PEDT score of the APE patients was correlated negatively with the total IIEF-15 score (r=－0.391, P<0.01) and the scores in the domains of erectile function (r=－0.362, P<0.01) and overall satisfaction (r=－0.621, P<0.01), but not correlated with intercourse satisfaction, sexual orgasm, or sexual desire. The PEDT score of the LPE group was correlated negatively with intercourse satisfaction (r=－0.286, P<0.05) but not correlated with either the total IIEF-15 score or the scores in the domains of erectile function, overall satisfaction, sexual orgasm, or sexual desire. CONCLUSIONS: PE patients have a higher PEDT score and a lower IIEF-15 score than normal males. The PEDT score of APE patients is significantly correlated with the total IIEF-15 score, while that of LPE patients is correlated not with the total IIEF-15 score but with intercourse satisfaction.

[Mechanism about LMP1 of EB Virus Promoting Plasma Blast Differentiation of DLBCL Cell via mTORC1].

OBJECTIVE: To investigate possible mechanism on protien LMP1 expressed by EBV inducing plasmablast differentiation of DLBCL cell via the mTORC1 pathway. METHODS: The expression levels of LMP1 protein, CD38 and the phosphorylation levels of p70S6K in EBV+ and EBV- DLBCL cell lines were detected by Western blot. Cell lines overexpressing LMP1 gene stablely were constructed and LMP1 gene was silenced by RNAi. The expression of LMP1 gene was verified by RT-qPCR. The expression levels of LMP1 and CD38 and the phosphorylation levels of p70S6K in each group were detected by Western blot. RESULTS: Compared with EBV-DLBCL cells, the expression of LMP1 was detected on EBV +DLBCL cells (P =0.0008), EBV +DLBCL cells had higher phosphorylation levels of p70S6K (P =0.0072) and expression levels of CD38(P =0.0091). Compared with vector group, the cells of LMP1OE group had higher expression levels of LMP1 and CD38 (P =0.0353; P <0.0001), meanwhile molecular p70S6K was phosphorylated much more(P =0.0065); expression of LMP1 mRNA was verified(P <0.0001). Compared with si-NC group, expression level of LMP1 protein(P =0.0129) was not detected and phosphorylated p70S6K disappeared of LMP1KO group (P =0.0228); meanwhile, expression of CD38 decreased,although there was no significant difference (P =0.2377). CONCLUSION: LMP1 promotes DLBCL cells plasmablast differentiation via activating mTORC1 signal pathway.

[Premature ejaculation and female partners' psychological factors].

Premature ejaculation (PE) is a most common sexual dysfunction in males, which variably affects men's reproductive and mental health. Recent studies show that PE affects not only the patients themselves, but also the psychological state of their spouses, which, in turn, may alleviate or aggravate the patients'PE symptoms by different reactions. However, the correlation between PE and female's psychological factors is mostly ignored by the patients and even clinicians in the studies, diagnosis and treatment of PE.

[Analysis on "the essence of tuina, arrival of qi ensuring curative effect"].

In reference with the systematic review of the thought of deqi (arrival of qi) put forward in Huangdi Neijing (Internal Classic of Yellow Emperor) and other classic books of traditional Chinese medicine, in view of detecting qi and identifying qi before treatment, as well as the prerequisites of deqi in tuina, meaning the accurate syndrome differentiation and manipulations, the importance of deqi in treatment with tuina is expounded. In association with clinical experience, the specific manifestations of deqi in patients during tuina are summarized, e.g. soreness, distention, pain, numbness, warm feeling and slight sweating, local changes in intestinal sound and skin color, as well as mind regulation. It is anticipated that deqi of tuina may be drawn the attention in clinical practice, and the relevant study be expanded.

Prevalence of insomnia and anxiety among healthcare workers during the COVID-19 pandemic in Jilin Province.

The outbreak of the 2019 novel coronavirus disease (COVID-19) has impacted the mental health of healthcare providers at the frontline. Therefore, we conducted this study to estimate the prevalence rate of anxiety and insomnia and identify associated risk factors among healthcare workers in Jilin, China, during the period from January 25 to February 25, 2020. Zung's Self-Reported Anxiety Scale (SAS) and the Insomnia Severity Index (ISI) scale were used to diagnose anxiety and insomnia, respectively. Associated risk factors were identified through a multivariate logistic regression model. A total of 300 healthcare workers were invited and 236 completed the study. Of them, 234 (99.15%) were medical workers, 197 (83.47%) were working at frontline departments, and 159 (67.37%) were fighting against COVID-19. Fifty-seven respondents (24.15%) had anxiety (SAS index score ≥45) and 94 (39.83%) had insomnia (ISI score ≥8). Based on the multivariate analysis, contact with people from Hubei province during work (no vs not clear) [OR=0.25, 95%CI: 0.10-0.61] and personal protective equipment (PPE) (not in place vs in place) [OR=6.22, 95%CI: 2.23-17.40] were significantly correlated with anxiety. PPE (not in place vs in place) was the only significant risk factor of insomnia [OR=10.56, 95%CI: 4.00-27.87]. The prevalence of anxiety and insomnia was high in our study, reflecting the psychological impact of COVID-19 on healthcare workers. The unavailability of PPE in place was a significant risk factor of both anxiety and insomnia.

Iodine-imine Synergistic Promoted Povarov-Type Multicomponent Reaction for the Synthesis of 2,2'-Biquinolines and Their Application to a Copper/Ligand Catalytic System.

An efficient iodine-imine synergistic promoted Povarov-type multicomponent reaction was reported for the synthesis of a practical 2,2'-biquinoline scaffold. The tandem annulation has reconciled iodination, Kornblum oxidation, and Povarov aromatization, where the methyl group of the methyl azaarenes represents uniquely reactive input in the Povarov reaction. This method has broad substrate scope and mild conditions. Furthermore, these 2,2'-biquinoline derivatives had been directly used as bidentate ligands in metal-catalyzed reactions.

[Relationship between PANTR1 and Imatinib Resistance of Chronic Myeloid Leukemia Cell Line K562 and Its Related Mechanisms].

OBJECTIVE: To investigate the relationship between long non-coding RNA (LncRNA) PANTR1 and imatinib resistance in chronic myeloid leukemia cell line K562 and its mechanism. METHODS: K562 control cells (Control) and K562 imatinib resistant cells (ImR) were cultured. Two siRNA vectors targeting PANTR1 and control vectors were transfected into K562-ImR cells by lentivirus as ImR-siPA#1, ImR-siPA#2 and ImR-siControl cells, respectively. Imatinib semi-inhibitory concentration (IC50) was detected by CCK-8 kit. The expression level of PANTR1, BCR/ABL, MDR, CD44 and CD133 mRNA were detected by fluorescence quantitative PCR（RT-qPCR）, and the expression level of BCR/ABL, MDR, CD44 and CD133 protein were detected by Western blot. RESULTS: Imatinib IC50 in ImR cells was significantly higher than that in control cells (P＜0.01), that of ImR-siPA#1 and ImR-siPA#2 cells were significantly lower than that in ImR-sicontrol cells (P＜0.01), but still significantly higher than that in control cells (P＜0.01). The mRNA expression level of PANTR1, BCR/ABL, MDR, CD44 and CD133 in ImR cells were significantly higher than those in control cells (P＜0.01). The mRNA expression level of PANTR1, MDR, CD44 and CD133 in ImR-siPA#1 and ImR-siPA#2 cells were significantly lower than those in ImR-siControl cells (P＜0.01), while the expression level of BCR/ABL mRNA was not significantly different (P＞0.05). The protein expression level of BCR/ABL, MDR, CD44 and CD133 in ImR cells were significantly higher than those in control cells, while the protein expression level of MDR, CD44 and CD133 in ImR-siPA#1 and ImR-siPA#2 were significantly lower than those in ImR-siControl cells. CONCLUSION: LncRNA PANTR1 can promote the expression of MDR and stem cell marker in chronic myeloid leukemia cell line K562, and mediate imatinib resistance.

[Serum Level of MiR-609 and Its Clinical Prognostic Value in Patients with Thalassemia].

OBJECTIVE: To investigate the level of serum microRNA-609 and its clinical prognostic value in patients with thalassemia. METHODS: One hundred and twenty-seven patients with thalassemia treated in our hospital from April 2017 to April 2018 were selected, 100 healthy persons were selected as control group. The changes of miR-609 were analyzed by RT-PCR, the relationship between miR-609 and clinical indicators of thalassemia was analyzed, and the prognostic risk factors of thalassemia were evaluated by multivariate logistic regression analysis. RESULTS: The relative expression level of miR-609 in thalassemia patients was 3.17±0.24, which was significantly higher than that in control group (P<0.05). The levels of ALT, Plt and MCH in patients with high expression of miR-609 were significantly higher than those in patients with low expression of miR-609 (P<0.05). The levels of Hb and sICAM-1 in patients with high expression of miR-609 were significantly lower than those in patients with low expression of miR-609 (P<0.05). There was no correlation between the level of miR-609 and the patient's sex, age and AST (P>0.05). The incidence rate of mild anemia in high expression group was significantly lower than that in low expression group (P<0.05). There was no correlation between the level of miR-609 and the incidence rate of moderate anemia (P>0.05). The number of patients with severe anemia in the miR-609 high expression group was higher than that in miR-609 low expression group (P<0.05). The incidence rate of dizziness, fatigue and fever in patients with miR-609 high expression group was significantly higher than those in patients with miR-609 low expression (P<0.05). There was no correlation between the level of miR-609 and the incidence rate of nausea in patients with thalassemia. ROC curve showed that the AUC value of microRNA-609 was 0.862, the sensitivity was 83.6%, and the specificity was 84.1%, which suggested that miR-609 had a high diagnostic value for thalassemia. Multivariate logistic regression analysis showed that MCH and mir-609 were risk factors for poor prognosis of thalassemia patients. CONCLUSION: The increased level of serum miR-609 in patients with thalassemia is a risk factor for poor prognosis and can be used as a reference index for evaluating the efficacy for patients.

Selective detection of superoxide anion radicals generated from macrophages by using a novel fluorescent probe.

Quantitation of superoxide radical (O (2)(-).) production at the site of radical generation remains challenging. A simple method to detect nanomolar to micromolar levels of superoxide radical in aqueous solution has been developed and optimized. This method is based on the efficient trapping of O(2)(-). using a novel fluorescent probe (2-chloro-1,3-dibenzothiazolinecyclohexene), coupled with a spectra character-signaling increase event. A high-specificity and high-sensitivity fluorescent probe was synthesized in-house and used to image O(2)(-). in living cells. Better selectivity for O(2)(-). over competing cellular reactive oxygen species and some biological compounds illustrates the advantages of our method. Under optimal conditions, the linear calibration range for superoxide anion radicals was 5.03 x 10(-9)-3.33 x 10(-6) M. The detection limit was 1.68 x 10(-9) M. Fluorescence images of probe-stained macrophages stimulated with 4beta-phorbol 12-myristate 13-acetate were obtained successfully using a confocal laser scanning microscope.

[Comparison of Laboratory Indexes and Therapeutic Efficacy between Two Kinds of Different RBC Transfusion in AIHA Patients].

OBJECTIVE: To investigate the effect of 2 kinds of red blood cells (RBC) on the laboratorial indexes and therapeutic efficacy of patients with autoimmune hemolytic anemia (AIHA). METHODS: The clinical data of 120 patients with AIHA from June 2015 to June 2016 were analyzed retrospectively. These 120 patients were divided into A goup and B group. The patients in A group (60 cases) were infused with washed RBC, while the patients in B group (60 cases) were infused with WBC-deplated RBC. The changes of laboratotial indexes, clinical symptoms and incidence of adverse reactions in 2 groups before treatment and at 24 hourse after treatment as well as the therapeutic efficacy and improvement status of clinical symptoms were compared. RESULTS: The RBC count and Hb level in 2 groups after treating for 1 day were significantly higher than those before treatment (P<0.01), while the TBIL level and reticulocyte (Ret) count were significantly lower than those before treatment (P<0.01). However, the RBC, Hb, TBIL levels and Ret count in 2 groups before and after treatment showed no statistical difference (P>0.05); the improvement of clinical symptoms, complex therapeutic efficacy and incidence of adverse reactions in 2 groups after treatment also showed no significantly difference (P>0.05). CONCLUSION: Both washed and leukocyte-deplated RBC can alleviate the anemia in patients with AIHA in short-time, but the leucocyte-deplated RBC is more expensive, suggesting that the wased RBC is more practical for treatment of AIHA patients.

[Analysis of Irregular Antibody Production in Severe ß-Thalassemia Children with long-term Blood Transtusion and Its Correlation with RH Gene and Anemia Gene Mutation Site].

OBJECTIVE: To analyze the prevalence of irregular antibodies in children with severe ß-thalassemia after long-term blood transfusion and its correlation with RH and anemia gene mutations site. METHODS: One hundred twenty children with severe ß-thalassemia and long-term blood transfusion were selected in our hospital, the genomic DNA was extracted and the genotype of RH factor were determined by PCR-SSP. The irregular antibodies and their types were screened and identified by the serological method, the gene types of the severe ß-thalassemia were analyzed by reverse dot blot hybridization on DNA chip and PCR amplification. RESULTS: The major of RH genotypes in 120 children were Ce/Ce (59.17%) and CE/ce (25%), among them 10 children possessed the positive irregular antibodies (8.33%), out of these 10 children, the genotypes of RH factor were Ce/Ce in 6 cases, cE/cE, CE/ce, cE/ce and Ce/ce in one case; among these 10 children with positive irregular antibodies, the anemia gene mutations were IVs-11654M in 2 cases, cD4142M in 6 cases, 28M in 1 case, and CD71-72M in 1 case. CONCLUSION: Irregular antibodies produced by regular blood transfusion in children with severe ß-thalassemia may be related with RH factor and anemia gene mutation sites.

Determination of trace hydroxyl radicals by flow injection spectrofluorometry and its analytical application.

On the basis of the fluorescence increase of the reaction of ninhydrin with hydroxyl radicals, a new method for the determination of trace amounts of hydroxyl radicals by flow injection spectrofluorometry is presented. The introduction of flow injection analysis brought better reproducibility and avoided the effect of oxygen and other substances in the environment on the reaction of ninhydrin with hydroxyl radicals. Under optimum experimental conditions, the hydroxylated product of ninhydrin had excitation and emission maxima at 300 and 406 nm, respectively. The linear range was 2.60 x 10(-7) to 4.00 x 10(-5) M, and the limit of detection was 7.91 x 10(-8) M. A high analysis rate of 22 samples per hour was obtained. The proposed method has been applied successfully to the determination of scavenging effects of thiourea and vitamin C on hydroxyl radicals as well as to the evaluation of antioxidant capacities of some natural food.