1.The effects of CoCl2-induced in vitro hypoxia on aquaporin-4 expression in cultured astrocytes
Hong LU ; Hui HU ; Zhanping HE
Chinese Journal of Neurology 2011;44(2):117-121
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.
4.The minimally invasive surgery for treating recurrent lumbar disc protrusion
Hui WU ; Lingyun HU ; Hong LIN
Chongqing Medicine 2014;(27):3556-3558
Objective To study the clinical effect of the minimally invasive surgery for treating recurrent lumbar disc protrusion . Methods Thirty operative cases of recurrent lumbar disc protrusion treated with the minimally invasive surgery from July 2009 to July 2013 were retrospectively analyzed for statistically describing the age ,body mass index ,recurrent segment ,operating time ,intr-aoperative bleeding volume and postoperative painless walking time .The recurrence segments were 1 case of L34 ,10 cases of L45 and 19 cases of L5 S1 ;the visual analogue scale (VAS) was adopted to evaluate the operative effect .Results The operating time was 65 min on average (50-100)min;mean intraoperative bleeding volume was 145 mL(100-180)mL ;average length of hospital stay was 13 d (9-16)d;average hospitalization costs were 7 300 Yuan;average postoperative painless walking time was 11 d (7-15)d;post-operative non-manual labor work time was 27 .3 d .The VAS score was reduced from (7 .3 ± 1 .3) before operation to (3 .1 ± 0 .9) after operation and (2 .2 ± 0 .6) at the last follow-up ,which was statistically significant(P<0 .05) .Followed up for 6-33 months , average 16 .5 months .Superior efficacy in 16 cases ,gord in 7 cases ,average in 5 cases ,bad in 2 cases ,the rate of good efficacy was 76% .Conclusion The minimally invasive surgery is one of effective methods to treat postoperative recurrence of lumbar disc pro-trusion .
5. Influence of cooling rate on qualtiy of cryopreserved platelets
Academic Journal of Second Military Medical University 2010;29(7):809-812
Objective: To observe the outcomes of different cooling rates in the cryopreservation of platelets, so as to screen the best cooling rate for cyropreservation of platelets. Methods: Platelet concentrate was cooled down with the new cryopreserving solution at various cooling rates(1, 10, 20°C/min) and was kept at ∼80°C for one week. Then platelets were thawed to check platelet count, adrenaline induced aggregation, and expression of CD41, CD42b and CD62p; the results were compared with those of the non-programmed cooling group and fresh platelets. Results: The platelet counts of cryopreserved groups were lower than that of fresh platelet concentrate (P<0.05 or P<0.01). There were no differences in adrenaline induced aggregation between 10°C/min group, 1 °C/min group, and non-programmed cooling group; and the aggregation of the former 3 groups was lower than that of 1 °C/min group(P<0.01) and higher than that of 20 °C/min group (P<0.05). The CD42b expression in 10 °C/min group was higher than in other groups (P<0.05) and lower than in fresh platelet concentrate. The CD62p expression in 1 °C/min group, 20 °C/min group, none programme cooling group was higher than in fresh platelet concentrate (P<0.05). The CD62p ex: ression in 10 °C/min group was higher than other 3 groups (P<0.05) and lower than in fresh platelet concentrate, with no statistical significance. Conclusion. Cooling rate has great impact on the quality of cryopreservted platelets. Compared with 1 °C/min and 20 °C/min, 10 °C/min is the optimal cooling rate with the new cryopreservation solution in this study.
6.Effects of E23K polymorphism in KCNJ11 gene on membrane current.
Xiao-Hui XIA ; Ai-Hong YANG ; Yang HU
Chinese Journal of Applied Physiology 2014;30(1):23-26
OBJECTIVEE23K polymorphism in KCNJ11 gene is associated with cardiovascular disease and diabetes. In order to explore the mechanism of E23K correlation to related diseases, the effect of E23K polymorphism in KCNJ11 gene on membrane current was investigated.
METHODSThe exon of KCNJ11 was obtained by PCR amplification and the G-->A mutation was completed by overlap extension PCR. The sequences of KCNJ11 exon contained 23E or 23K was inserted into pcDNA3.1/CT-GFP vector respectively. The recombinant plasmid, pcDNA3.1-KCNJ11(E) and pcDNA3.1-KCNJ11(K), were transfected into HEK293T cells by lipofectamine and the membrane current density was determined by whole-cell patch clamp technique.
RESULTS1,173 bp sequences of KCNJ11 gene's exon were amplified by PCR and the recombinant expression plasmid, pcDNA3.1-KCNJ11(E) and pcDNA3.1-KCNJ11(K), were constructed successful. Positive and negative currents were detected in HEK293T cells transfected with difference plasmid by whole-cell patch clamp technique. Results showed that the reversed voltage was 50mV. The current in HEK293T cells with pcDNA3.1-KCNJ11(E) was significantly greater than that with pcDNA3.1-KCNJ11(K) (P < 0.05, n = 10).
CONCLUSIONThe polymorphism of E23K in exon of KCNJ11 gene changed the membrane currents in HEK293T cells. It could be an experiment support for the possible mechanism between the locus and related diseases.
Exons ; HEK293 Cells ; Humans ; Membrane Potentials ; Patch-Clamp Techniques ; Polymorphism, Genetic ; Potassium Channels, Inwardly Rectifying ; genetics
7.Therapic effect of lauromacrogol sclerotherapy on infantile hemangioma
Shan JIANG ; Ling LU ; Yingzi HU ; Hui LI ; Hong LIANG
Chinese Journal of Medical Aesthetics and Cosmetology 2013;19(5):359-361
Objective To investigate the effect of intralesional lauromacrogol therapy on infantile hemangioma.Methods 30 cases all received intralesional injection of lauromacrogol.The changes of tumor size,texture and color were monitored and recorded.The adverse effects after medication were observed and managed accordingly.The adverse effects after medication were observed.The times of treatment were determined according to degree of reduction of tumor volume.To assess the efficacy,a 4 scales system was adopted.Results All patients had completed the treatment and followup.The overall response was scale Ⅰ in 2 cases (6.7 %),scale Ⅱ efficacy in 4 patients (13.3 %),scale Ⅲ efficacy in 5 cases (16.7 %) and scale Ⅳ in 19 cases (63.3 %).The smaller the tumor,the fewer the times of treatment,and the better the results.The effects of treatment were better if patient could be treated by intralesional lauromacrogol therapy in early stage.No severe adverse effects were observed during 6 months of follow-up.Conclusions Intralesional injection of lauromacrogol sclerotherapy is a safe and effective way to treat infantile hemangioma.
8.Expression of aquaporins 4 and pathological changes in early phase of traumatic brain edema
Hong LU ; Xiaoyan LEI ; Hui HU ; Zhanping HE
Chinese Journal of Trauma 2013;29(12):1224-1229
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.
9.Alagille syndrome in a child combined with hepatocellular carcinoma:a case report and literature review
Hui HU ; Yongchen YANG ; Hong ZHANG ; Ting ZHANG ; Yongmei XIAO
Journal of Clinical Pediatrics 2017;35(4):253-255
Objective To explore the clinical manifestation, imaging examination, treatment and prognosis of Alagille syndrome in a child combined with hepatocellular carcinoma. Method The clinical manifestation, assistant examination and diagnosis of Alagille syndrome combined with hepatocellular carcinoma were analyzed in the child, and the pertinent literature were reviewed. Results The 6-year-old girl was admitted to hospital for repeated jaundice, and had a history of cardiac surgery. After admission, the patient was found to have a typical face look such as frontal bossing, sunken eyes, pointed chin and hypertrophy of nasal tip. Blood biochemistry showed intrahepatic cholestasis and increased alpha-fetoprotein. Abdominal ultrasonography revealed diffuse multiple solid lesions in the liver. And magnetic resonance imaging of the liver indicated that the liver was enlarged and multiple solid space occupying masses. Jagged 1 gene detection showed heterozygosis mutation of c.1205delC. Conclusion Alagllie syndrome complicated with hepatocellular carcinoma in childhood is extremely rare, and early diagnosis and long-term follow-up are of positive significance for its treatment and prognosis.
10.Surgical treatment of cervical esophageal carcinoma
Shi-Xin ZHENG ; Hui-Ge WANG ; Hong-Yi HU ;
Chinese Journal of Postgraduates of Medicine 2006;0(09):-
Objective To explore the application of gastric(enteric)-pharyngeal anastomosis for cervical esophageal carcinoma. Methods The clinical data of 12 cases with surgical management of cervical esophageal carcinoma were retrospectively analyzed. Results The resectability of cervical esophageal carcinoma was 100%,no case complicated with pharyngeal fistula.Swallowing function of all cases was in a good state.The overall follow-up was 1 to 7 years,among them 9 surviving,3 dead.The surviving 5 cases are over 3 years,the ongest beyond 7 years. Conclusions Gastric(enteric)-pharyngeal anastomosis is a good primarily rehabilitating method of the cervical esophageal defect after surgical treatment of cervical esophageal carcinoma.