Objective To investigate the expression of aquaporin-4 (AQP4) in cultured astrocytes after in vitro hypoxia induced by CoCl2. Methods After primary culture and subculture, the astrocytes were placed in a controlled atmosphere culture chamber. Both control group and hypoxia groups were established.These groups were further divided into seven sub-groups according to the different time intervals: 15, 30minutes and 1,2, 4, 6, 12 hours, respectively (6 apertures for each group). The shape of the astrocytes in each group was observed with light microscopy and transmission electron microscopy ( TEM ). All groups were examined using in situ hybridization, real time fluorescence quantitative reverse transcriptase polymerase chain reaction, immunocytochemistry and Western blot. The data was analyzed statistically with SPSS 13.0 software. Results There was significant consistency between the AQP4 mRNA and protein ( r =0. 85, P ＜0. 01 ). There was slight positive expression of AQP4 in a few astrocytes of the control groups. In the hypoxia groups, the expression of AQP4 increased within 15 minutes; the increase was most prominent between 1 and 4 hours( mRNA in hypoxia groups: 0. 26 ± 0. 04, 0. 31 ± 0. 02, 0. 36 ± 0. 04; control groups:0. 06 ±0. 01,0. 09 ±0. 01,0. 08 ±0. 01 )after hypoxia and became less between 6 and 12 hours; There was significant difference in the AQP4 expression between the hypoxia groups and control groups among different time points (t = 16. 51, 18.20, 15.26,all P＜0. 01 ). The corresponding pathological changes were cellular edema, which was most prominent between 1 and 4 hours. Under TEM, increase in size of the nucleolus and swelling of endoplasmic reticulum and mitochondria; these changes became more marked with time.Disruption of a few astrocytes was detected in the hypoxia groups at 12 hours. Conclusions The pathological change of astrocytes is cellular edema following hypoxia. There is a positive relationship between the presence and degree of cellular edema as well as the duration of hypoxia and the up-regulating of AQP4.These results imply that AQP4 expression is an important molecular mechanism of celluar edema of astrocytes.

Hoxa3-Pax1/Pax9-Eya1-Six1/4 regulatory pathway seems to be operating during forming the bilateral parathyroid/thymus common primordial in early embryonic development.The specification of the parathyroid domain in the parathyroid/thymus primordial is regulated through a Shh-Tbx1-Gcm2 pathway.Gcm2 also may play roles in later steps of parathyroid development,including CaSR and PTH gene expression.MafB and Gcm2 interact with each other and synergistically activate PTH transcription.Genetic basis and the etiology of some hypoparathyroid disorders in man are involved defects in transcription factors that include GCMB,GATA3,Tbxl,SOX3 and GNA11.This marker expression in thymus and parathyroid primordium includes HoxA3,Pax1,Eya1,and Six1;and expression of parathyroid cell-like cells includes Gcm2,CaSR,and PTH.These expressions may serve as markers of stem cell differentiation into parathyroid cell-like cells.

Air Pollutants ; toxicity ; Air Pollution, Indoor ; adverse effects ; Animals ; DNA Damage ; drug effects ; Female ; Interior Design and Furnishings ; Male ; Mice ; Mutagenicity Tests

3' Untranslated Regions ; genetics ; 5' Untranslated Regions ; genetics ; Enterovirus B, Human ; genetics ; Genome, Viral ; genetics ; Humans ; Poly A ; genetics

Objective To optimize in vitro amplification of human γδ T cells with cytokines for tumor adoptive immunotherapy.Methods On the basis of the immobilized anti-TCR γδ antibody plus IL-2 system, other γ chain receptor family cytokines, including IL-7, IL-15 and IL-21, were tested to amplify human peripheral blood γδ T cells either alone or in diversity combination.The percentage of γδ T cells was measured by flow cytometry, and the proliferation efficiency of γδ T cells was calculated.The expression of proliferation-or cytotoxicity-related molecules on γδ T cells was examined by flow cytometry in order to explore the relevant mechanisms.The cytotoxicity of γδ T cells to Daudi cells was detected by lactate dehydrogenase.Results IL-15 alone but not IL-7 or IL-21 increases the γδ T cell purity, amplification efficiency and cytotoxicity to reach comparable levels to those of IL-2.IL-2 plus IL-15 up-regulates the expression of CD69 on γδ T cells and significantly increases their amplificationefficiency (P<0.05).IL-2 plus IL-21 enhanced the cytotoxicity of γδ T cells against Daudi cells by increasing the expression of granzyme A (P<0.001).The combination of IL-2, IL-15 and IL-21 significantly improves cytotoxicity of γδ T cells but reduces their amplification efficiency.In addition, when IL-21 was applied for a short time, it also enhanced the cytotoxicity of γδ T cells (P<0.05).Conclusions The combination of IL-2 and IL-15 as well as a short time addition of IL-21 is the best cytokine recipe to amplify human peripheral blood γδ T cells in vitro with immobilized anti-TCR γδ antibody, which can increase both the proliferation efficiency and the cytotoxicity to tumor cells of γδ T cells.

Objective To investigate the expression of aquaporins 4 (AQP4) and histopathological changes in early phase of traumatic brain edema and the correlation between AQP4 expression and structural damage to blood-brain barrier (BBB).Methods A total of 120 healthy adult Wistar rats were divided into sham operation group and brain trauma group (which was subgrouped at hours 1,3,6,12 and 24 postinjury) according to random number table,with 20 rats per group.At each time point,brain water content was measured; brain edema and BBB structural changes were observed pathologically;IgG and AQP4 expressions in traumatic brain tissues were detected with immunohistochemical method and Western-blotting.Results In sham operation group,negatively stained IgG was observed and there were no abnormalities in brain tissue structure,brain water content as well as AQP4 expression.In brain trauma group,cerebral water content presented notable increase at 6 hours postinjury and peaked at 24hours; IgG expression showed significant increase at 1 hour postinjury,peaked at 6 hours postinjury and remained a high level at 24 hours.Pathologic observation revealed damage to BBB,blood red cells leaking out of the blood vessels,and tissue gap widening at 1 hour postinjury,which manifested as vasogenic brain edema.Further,those phenomena were gradually aggravated over time and became obvious at 6 hours postinjury.Intracellular edema occurred at 3 hours postinjury,with the presence of increased glial cell body,cytoplasm light staining or vacuolar degeneration,as well as mitochondria swelling and was also aggravated with time,particularly at 6 hours postinjury.Except that the previously mentioned two forms of edema coexisted at 12 hours postinjury,tissue necrosis,inflammatory cell infiltration and microglia proliferation were emerged and aggravated at 24 hours postinjury.AQP4 level decreased at 1 hour,minimized at 6 hours and regained at 12 hours,showing a V-shape curve.Conclusions Vasogenic edema characterized by BBB disruption is the primary histopathological change in early-phase of brain trauma,followed by the coexistence with intracellular edema and aggravation of the two forms of edema over time.AQP4 expression is down-regulated in the vasogenic edema phase but highly expressed at phase of the intracellular edema.

Objective To determine if the cardiac index (CI) measured with arterial pressure-based cardiac output (APCO) agrees with that measured with pulmonary artery catheter (PAC) during cesarean section in parturients with congenital heart disease and severe pulmonary hypertension.Methods Thirty parturients with congenital heart disease and severe pulmonary hypertension,scheduled for elective cesarean section,were enrolled in the study.APCO and PAC were used for cardiac output and other parameters of hemodynamics monitoring during operation in all patients.Continuous epidural anesthesia was performed.CI was recorded immediately before epidural administration (T1),at 5 and 10 min after epidural administration (T2-3),immediately before delivery (T4),and at 2 and 5 min after delivery (T5-6).The correlation was tested by Pearsone correlation analysis.CI derived from APCO and PAC was compared by Bland-Altman analysis for agreement.Results CI measured with PAC was significantly higher than that measured with APCO (P ＜ 0.05).Compared with the baseline value at T1,no significant difference in CI measured with PAC and APCO was found at T11-T4 and T6 (P ＞ 0.05),and CI measured with PAC and APCO was significantly increased at T5 (P ＜ 0.05).Bland-Ahman analysis showed poor agreement between CI measured with the two methods.The correlation analysis showed that there was positive correlation between CI derived from the two methods at T1-6 (the correlation coefficient was 0.93,0.95,0.94,0.89,0.96 and 0.91 at T1-6,respectively,P ＜ 0.05).Conclusion CI value obtained with APCO agrees poorly with that obtained with PAC during cesarean section in parturients with congenital heart disease and severe pulmonary hypertension,but agrees well in monitoring the changing trend of CI.

Objective: To investigate the potential role of me latonin in the adrenal cortex of human embryo. Methods:Specifi c melatonin receptors was localized and characterized in the adrenal cortex of h u man embryo by means of immunohistochemistry. Results: mt1 （Me l1a）and MT2 （Mel1b）subtype of melatonin receptors was principally localize d to cytoplasm in zona glomerulosa， fasciculata and reticularis. Conclu sion: It is possible that mt1 and MT2 subtype of melatonin receptors co-exist in the adrenal cortex of human embryo.

AIM:To investigate the safety and effect of femtosecond laser-assisted cataract surgery with Cionni modified capsular tension ring (MCTR) implantation in the management of traumatic lens subluxation.METHODS: Totally 11 patients (11 eyes) with traumatic lens subluxation were divided into three groups according to the severity of lens dislocation, ranging from 90° to 120° (4 eyes), 120° to 180° (5 eyes) and 180° to 270° (2 eyes).The contact LenSx femtosecond laser cataract surgery platform was applied to create the capsulotomy, prepare nuclear fragmentation and make corneal wound creation.Anterior vitrectomy was performed in some patients during the surgery.After capsular retractors insertion and phacoemulsification, the MCTR was inserted to the capsular bag and fixed to the sclera.Finally, the IOL was implanted into the capsular bag.Postoperative visual acuity, intra-and post-operative complications, anterior capsular opening, IOL and MCTR position and intraocular pressure (IOP) were assessed.RESULTS:The duration of follow-up was 2mo.All the operations were completed successfully.Five eyes underwent cataract surgery combined with anterior vitrectomy.Four eyes had been inserted with 2-eyelet MCTR and seven eyes with 1-eyelet MCTR.The best corrected visual acuity (BCVA) after operation was better than 0.5 in 4 eyes, between 0.3 and 0.5 in 3 eyes, between 0.1 and 0.3 in 3 eyes, and less than 0.1 in 1 eye.Compared with preoperative BCVA, the difference was statistically significant (P<0.05).All the IOLs were stably centered and the eyelet of MCTR was fixated steadily between the iris and the anterior capsule.The common intra-and post-operative complications were subconjunctival hemorrhage, incomplete capsulotomy, residual cortex, secondary glaucoma and posterior capsular opacification.CONCLUSION:Femtosecond laser-assisted cataract surgery can improve the success rate of capsulorhexis, and reduce the difficulty of nuclear fragmentation.Femtosecond laser-assisted cataract surgery combined with MCTR implantation is an ideal surgical method for traumatic lens subluxation.