Objective To construct an immortalized rat astrocyte strain expressing enkephalin which can be regulated by doxycycline.Methods pRevTet-On and recombinant plasmid pRevTRE/hPPE were transfected into packaging cell PT67 respectively by standard lipofectamine methods.The supernatant containing RevTet-On virus was used to infect immortalized rat astrocyte strain(IAST)and RevTRE/hPPE virus was used to infect the IAST/Tet-On cell.Immortalized rat astrocyte strain that stably expressed Tet-regulated enkephalin was established. hPPE gene expression level of IAST/Tet-On/hPPE cell strain was detected by real time-PCR,immuno- cytochemistry and radio-immunoassay.Results Real time-PCR,immuno-cytochemistry and radio-immunoassay confirmed the level of enkephalin expression was regulated by doxycycline in a dose-dependent manner.Conclusion An immortalized rat astrocyte strain which secrets enkephalin as controlled by doxycycline has been constructed successfully.

Acupuncture Therapy ; Diaphragm ; pathology ; Humans ; Male ; Middle Aged ; Respiratory Paralysis ; pathology ; therapy

Objective To investigate the value of measurement of cardiac output in children by bio-reactance versus echocardiography.Methods Pediatric patients admitted in pediatric department of Peking University First Hospital from September to December 2012 who needed hemodynamic monitoring were enrolled prospectively.Cardiac index(CI)and stroke volume(SV)were measured by echocardiography and non-invasive cardiac output measurement(NICOM)and compared by Spearman correlation and Bland-Alt-man analysis.Results Thirty patients were included.The median age was 7.25 years.CI[M(P5 ,P95 )] measured by NICOM and echocardiography were correlated significantly[3.42(2.28,4.92)L /(min?m2 ) vs.3.51 (2.94,4.85 )L/(min?m2 ),R =0.385,P =0.035 ].Bland-Altman analysis revealed a bias of-0.22 L/(min?m2 )(P =0.051 ),limits of agreement of -1.40 to 0.95 L/(min?m2 ).SV[M(P5 ,P95 )] measured by NICOM and echocardiography were correlated more significantly [36.3 (12.6,87.8 )ml vs.39.4(14.7,86.9)ml,R =0.768,P ﹤0.001 ].Bland-Altman analysis revealed a bias of -3.1 ml(P =0.176),limits of agreement of -27.4 to 21.2 ml.Conclusion There is no significant difference between NICOM and echocardiography for the measurement of CI and SV in pediatric patients.Further validation studies need to be conducted before routine clinical use.

Mean hemoglobin (Hb) concentration of about 3 500 subjects derived from 17 studies of Himalayan highlanders (Tibetans, Sherpas, and Ladakhis) was compared with lowlanders (Chinese Han, Indian Tamils) lived in the Himalayas, and European climbers during Everest expeditions as well as Andean natives. The results found that Hb concentration in Himalayan highlanders was systemically lower than those reported for Andean natives and lowland immigrants. These comparative data demonstrated that a healthy native population may successfully reside at high altitude without a significant elevation in Hb, and the lower Hb levels of Himalayan highlanders than those of migrated lowlanders and Andean natives are an example of favourable adaptation over the generations. In addition, excessive polycythemia has frequently been used as a marker of chronic mountain sickness (CMS). Altitude populations who have a higher Hb concentration also have a higher incidence of CMS. The low Hb in Himalayans suggested as showing adaptation over many generations in Tibetan stock. Recent work in Tibet, suggested that Tibetans there may have adapted to high altitude as a result of evolutionary pressure selecting for genes which give an advantage at altitude. All of the population genomic and statistical analysis indicated that EPAS1 and EGLN1 are mostly likely responsible for high altitude adaptation and closely related to low Hb concentration in Tibetans. These data supported the hypothesis that Himalayan highlanders have evolved a genetically different erythropoietic response to chronic hypoxia by virtue of their much longer exposure to high altitude.
Adaptation, Physiological ; Altitude ; Asian Continental Ancestry Group ; genetics ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Evolution, Molecular ; Hemoglobins ; genetics ; Humans ; Hypoxia-Inducible Factor-Proline Dioxygenases ; genetics ; Tibet

Adolescent ; Adult ; Aspergillosis ; complications ; microbiology ; Aspergillus ; isolation & purification ; Candida albicans ; isolation & purification ; Candidiasis ; complications ; microbiology ; Female ; Follow-Up Studies ; Granulomatosis with Polyangiitis ; complications ; microbiology ; pathology ; Humans ; Male ; Middle Aged ; Mucor ; isolation & purification ; Mucormycosis ; complications ; microbiology ; Nocardia ; isolation & purification ; Nocardia Infections ; complications ; microbiology ; Retrospective Studies ; Young Adult

AIM To observe whether acute stimulation of BRL 37344, a β3-adrenergic receptor (β3-AR) agonist, has the same effects of exacerbating hemodynamics and increasing in β3-AR expression on rats with failing heart as chronic administration. METHODSRats received two doses of isoprenaline (Iso, 340 mg·kg-1, sc, with a 24-h interval) to prepare heart failure model. After 8 weeks, rats were given iv BRL 37344 0.4 nmol·kg-1·min-1 for 10 min. Hemodynamics were measured at 0, 10, 30 min, 1, 2, 3 h, 1, 2 and 7 d after BRL 37344. Levels of β-AR mRNA in myocardium were measured at 0, 1, 2 and 7 d after BRL 37344 by reverse transcription-polymerase chain reaction. RESULTSCompared with Iso group, heart rate, left ventricular end systolic pressure and +dp/dtmax were significantly higher, and left ventricular end diastolic pressure was significantly lower during 1-3 h after BRL 37344 injection in Iso+BRL group. Then, they were restored to the same level as that prior to BRL 37344 injection. Compared with normal control, the levels of β1-, β2- and β3-AR mRNA displayed no significant change in BRL group;the level of β1-AR mRNA was lower and the level of β3-AR mRNA was higher in Iso group. In Iso+BRL group, much more lower β1-AR mRNA level and much higher β3-AR mRNA level were shown on d 2 and d 7 than Iso group. CONCLUSION Acute administration of β3-AR agonist has a shorter improved hemodynamics. But it caused the same result as chronic administration in reduction of β1-AR mRNA and increment of β3-AR mRNA in failure hearts, which may aggravate the cardiac functions.

Eleven old male patients with hyperuricemia were collected ( hyperuricemia group,65-90 years old ).10 healthy middle-aged males ( middle-aged group,30-40 years old) and 10 healthy old males ( older group 60-70 years old ) with normal blood uric acid level were used as controls.All of the subjects were given low purine content diet ( 250 mg/d ) for 3 days followed by high purine content diet ( 800 mg/d ) consecutively for another three days.The samples of fasting blood and 24 h urine were collected for assay.The results showed that there were no significant changes of serum uric acid ( UA ) concentration in three groups after low purine content diet.But the levels of serum UA in three groups all increased significantly after high purine content diet,and the change was higher in hyperuricemia group than middle-aged group [ ( 507.7 ± 108.1 vs 378.9 ± 80.1 ) μmol/L,P＜0.05 ].24 h urine uric acid excretion in three groups was all significantly decreased after low purine content diet and increased after high purine content diet.After high purine content diet,24 h urine uric acid was lower in hyperuricemia group than middle-aged group [ ( 2.99 ± 1.21 vs 3.62 ± 1.02 ) mmol/24 h,P＜0.05 ].Blood urea nitrogen levels in all subjects decreased after low purine content diet and increased after high purine content diet ( P＜0.05 or P＜0.01 ).Creatinine clearance rate in hyperuricemia group was decreased after high purine content diet compared with baseline [ (75.3 ± 20.3 vs 80.7 ±20.0) ml/min ],and there were no significant changes in other groups after low and high purine content diet.24 h urine protein in hyperuricemia group was higher than middle-aged group ( P＜0.05 ),and increased after high purine content diet with significant difference ( P＜0.05 ).These results suggest that high purine content diet and decreased by renal uric acid clearance mainly contribute to hyperuricemia in old people.

AIM and METHODS: To study the culture met ho d of human neural stem/progenitor cells, these cells derived from human fetal fo rebrains were maintained and expanded in serum-free defined medium containing ba sic fibroblast growth factors (bFGF), epidermal growth factor (EGF) and B27. Whe n they formed neurosphere, these three factors and supplemented FBS were removed to induce differentiation. Cell were cultured for 12-14 d, then fixed for immun ocytochemistry examination. RESULTS: This period of expansion resulted in a 107-fold incre ase in this heterogeneous population of cells. Upon differentiation, they form n eurons, astrocytes and oligodendrocytes, the three main phenotypes in the CNS. CONCLUSION: These results demonstrate the feasibility of long-t erm in vitro expansion of human neural progenitor cells. The advantages of s uch a population of neural precursors for allogeneic transplantation, including t he ability to provide an expandable, well-characterized, defined cell source, ca n form specific neuronal or glial subtypes.

Objective: To evaluate effects of color matching of different cavosurface margins on the resin composites in vitro.Methods:Twenty extracted human premolars with an A 2 shade buccal surface were used in this study .Rectangular shaped cavities (3.0 mm depth, 2.0 mm width, 2.0 mm length) were prepared in the center of the buccal surfaces .The gingival and occlusal cavosurface margins were prepared to be either shoulder or bevel;the other cavosurface margins remained vertical .Ten teeth were filled with Clearfil AP-X (AP), the other ten with Clearfil Majesty (MJ) and light cured.The color difference at the cavosurface margin area was measured using a spectrophotometer ( CrystalEye ) and evaluated by 3 observers subjectively .The data were statistically analyzed using repeated measures ANOVA and Chi-square test .Results:When measured by CrystalEye , the color difference between the tooth and resin composite was reduced from the center of restoration to the cavosurface margin area .Both objective and subjective evaluations showed that for AP , the color difference at the cavosurface margin area had no statistical difference among 3 types of the margins; for MJ, the color difference at bevel margin area was significantly smaller than that at the vertical margin area .Conclusion: The resin com-posite restorations produced the color matching at marginal area .The color matching of resin composites with higher diffused light transmission property is more susceptible to the type of cavosurface margins . Preparing bevels may reduce the color difference between the restoration and tooth surface .

Objective To evaluate the effect of hypoxic-ischemic time on reduction of hypoxic-ischemic brain injury by sevoflurane postconditioning in neonatal rats.Methods Two hundred and ten 7-day-old Sprague-Dawley rats (105 male,105 female),weighing 13-17 g,were randomly divided into 7groups (n=30 each) using a random number table:sham operation group (group Sham),hypoxia-ischemia group (group HI),and sevoflurane postconditioning at different hypoxic-ischemic time point groups (P0,P3,P6,P 12 and P24 groups).Immediately after ligation of the left common carotid artery,and at 3,6,12 and 24 h after ligation,the rats inhaled the mixed gas containing 2％ sevoflurane for 30 min in P0,P3,P6,P13 and P24 groups,respectively.The fatality was recorded within 7 days after establishment of the model.At 7 days after establishment of the model,the rats were sacrificed,the brains were removed,and the right and left cerebral hemispheres were weighed separately,and the left/right cerebral hemisphere weight ratio was calculated.The hippocampal CA1 region and posterior cingulate gyrus were isolated,and the ratio of density of normal neurons in the left to the right was calculated.Results Compared with group Sham,the left cerebral hemisphere weight,left/right cerebral hemisphere weight ratio,and ratio of density of normal neurons were significantly decreased,and the fatality rate was increased in the other six groups (P＜0.05).Compared with group HI,the left cerebral hemisphere weight,left/right cerebral hemisphere weight ratio,and ratio of density of normal ncurons were significantly increased in P0,P3 and P6 groups (P＜0.05),and no significant change was found in the parameters mentioned above in P12 and P24 groups (P＞0.05).Compared with group P6,the left cerebral hemisphere weight,left/right cerebral hemisphere weight ratio,and ratio of density of normal neurons were significantly increased in P0 and P3 groups (P＜ 0.05).There was no significant difference in the parameters mentioned above between group P0 and group P3 (P＞0.05).Conclusion Sevoflurane postconditioning performed within 6 h of hypoxia-ischemia can reduce hypoxic-ischemic brain injury,and it provides no cerebral protection if exceeding 12 h.