0.05). Conclusion CCM is a proper scaffold,on which KC and FB of tympanum maintain satisfactory proliferation and collagen secretion activity.

Objective To study the effect of spontaneous subarachnoid hemorrhage (SAH)on transient receptor potential melastatin 4 (TRPM4)channel activity. Methods Seventeen SD rats of clean grade were selected. They were randomly divided into either a SAH (n = 10)or a sham operation group (n = 7) according to the random number table. At day 5 after SAH modeling,the cerebral arteries were harvested and the cerebral arterial smooth muscle cells were isolated using enzymatic digestion method. Western blot was used to detect TRPM4 expression and translocation rate. Patch-clamp techniques were used to study the maximum current intensity of the TRPM4 single channel in cerebral arterial smooth muscle cells. Results The fluorescent-stained TRPM4 were observed in cerebral arterial smooth muscle cells in the 2 groups of rats. The relative quantities of TRPM4 in the total protein of the sham operation group and the SAH group were 24 ± 3% and 32 ± 4% respectively. There was significant difference between the 2 groups (t = 4. 47,P < 0. 01). The translocation rates of TRPM4 in the sham operation group and the SAH group were 44. 0 ± 1. 9% and 60. 1 ± 2. 3% respectively,and the SAH group was higher than the sham operation group (χ2 = 4. 48,P < 0. 05). When the clamping voltages were - 100 mV,- 80 mV,- 60 mV,and - 40 mV,the maximum current intensity of TRPM4 single channel of the sham operation group was more than that of the SAH group. There were significant differences between the 2 groups (- 1. 90 ± 0. 10 mV vs. - 2. 23 ± 0. 08 mV,- 1. 68 ± 0. 12 mV vs. - 1. 99 ± 0. 12 mV,- 0. 89 ± 0. 09 mV vs. - 1. 24 ± 0. 09 mV,and - 0. 69 ± 0. 12 mV vs. - 0. 92 ± 0. 11 mV;all P < 0. 01). When the clamping voltages were - 20,0,20,40,60,80,and 100 mV,there was no significant difference in the maximum current intensity of TRPM4 single channel between the 2 groups (all P > 0. 05). Conclusion SAH has the induced effect for TRPM4 activity.

BACKGROUND:Primary human lung epithelial cel s are difficult to be isolated and cultured in vitro, which is characterized as limited sources, low cel viability, slow proliferation capacity, and lacking of differentiation capability.
OBJECTIVE:To establish an air-liquid interface model of lung epithelium by in vitro proliferation of human bronchiolar epithelial cel s, which is used for research on function of lung epithelial cel s.
METHODS:Primary human bronchiolar epithelial cel s were isolated using Pronase and DNase I combined digestive methods, and then proliferated using medium containing ROCK kinase inhibitor. The proliferated cel s were used for establishment of the air-liquid interface epithelium model. Cel differentiation was identified using scanning electron microscope, phase contrast microscope and immunofluorescent staining.
RESULTS AND CONCLUSION:The primary human bronchiolar epithelial cel s could be expanded successful y using medium containing ROCK kinase inhibitor, and the basal cel marker Cytokeratin14 was preferential y expressed in the proliferated cel population, indicating that these basal cel s might be the main subpopulation of human lung epithelial stem cel s. Subsequently, the proliferated cel s under the air-liquid interface could differentiate into ciliated cel s and non-ciliated column cel s. The results suggest that the proliferation and differentiation of human bronchiolar epithelial cel s were maintained in the presence of ROCK kinase inhibitor, and the air-liquid interface could promote the differentiation of human bronchiolar epithelial cel s.

Objective To investigate the effect of arterial and venous subarachnoid hemorrhage ( SAH)on voltage-dependent calcium channel( VDCC)currents of cerebral artery smooth muscle cells and the relationship between the concentration of oxyhemoglobin( OxyHb)in arterial and venous blood and cerebral blood flow. Methods Thirty-six clean grade rats were colleted. A rat SAH model was induced by injection of autologous arterial or venous blood in suprasellar cistern using assisted stereotaxic apparatus. The rats were divided into three groups:an arterial SAH( n=14 ),a venous SAH( n=13 ),and a sham operation( n=9 )group. The arterial and venous OxyHb concentrations were measured. Three days after SAH modeling,a patch clamp was used to detect the relative surface area of the cerebral artery smooth muscle cells,resting potential,and VDCC currents in rats. A fluorescent microsphere method was used to quantitatively analyze cerebral blood flow(CBF). Results (1)Arterial SAH OxyHb concentration (127 ± 4 g/L)was significantly higher than venous SAH OxyHb concentration(54 ± 6 g/L),and that of the sham operation group was 50 ± 5 g/L. The differences were statistically significant among the 3 groups( P<0. 01).(2)The maximum current of VDCC of the arterial SAH group(3. 22 ± 0. 31 pA)was significantly higher than that of the venous SAH group(2. 19 ± 0. 27 pA)and the sham operation group(2. 18 ± 0. 29 pA). The differences were statistically significant among the 3 groups( P<0. 01 ). The VDCC currents of the arterial SAH group were consisted of L- and R-currents,while the currents of the venous SAH group were only consisted of L-VDCC.(3)The cerebral blood flow of the arterial SAH group(0. 83 ± 0. 14 mL/[g·min])was significantly higher than that of the venous SAH group(1. 28 ± 0. 28 mL/[g·min])and the sham operation group(1. 35 ± 0. 19 mL/[g·min]). The differences were statistically significant(P<0. 01). Conclusions The changing effect of arterial SAH on the expression and function of the cerebral artery smooth muscle cells are greater than that of the venous SAH. This difference may be associated with the concentration and composition of vasospasm factors of OxyHb in arterial and venous blood.

To evaluate the efficacy of Tuina and Chinese patent drug Shuxuetong injection in preventing patients undergoing total knee arthroplasty from deep venous thrombosis and in functional rehabilitation.

Objective To summarize the experience of the therapy and diagnosis of thoracictuberculosis. Methods Diagnosis and operation of 163 cases of thoracictuberculosis were analyzed.Results 163 cases of thoracic- tuberculosis were treated with focuspurge upon two weeks' anti-tuberculosis treatment.153 cases were cured upon one operation.10 cases suffered incision delayed healing and there were no recurrence cases.Conclusion Thoraeictu- berculosis was treated with focuspurge upon two weeks anti-tuberculosis treatment before operation.Complete purge of focus and postoperative compression band and residual cavity filled with music flap were important measures to prevent incision delayed healing and recurrence.

Objective To evaluate the effect of iodine supplementation on improvement of developmental quotient (DQ) at the critical period of infant brain development.Methods Pregnant,lactating women and infants less than 3 years old were supplemented with iodized oil in Linxia Hui Autonomous Prefecture(Linxia Prefecture) Gansu Province in 2006-2010.Before and after the intervention(2006,2007-2010),five townships were randomly selected in the north,the south,the east,the west and the center of eight counties(cities) of Linxia.One village was chosen from each of those townships and 20 infants,20 pregnant women and 20 lactating women were randomly selected in each village(insufficient was made up from the neighboring villages).Urinary iodine(UI) level of the infants,pregnant and lactating women were determined by arsenic cerium catalytic spectrophotometry.DQ value of infants was measured before and after supplementation of iodized oil in 2006 and 2010.UI value of pregnant,lactating women and infants was monitored every year after iodine supplementation.Results Before iodine supplementation(2006),the median UI level of pregnant,lactating women and infants was 89.28,84.85,107.3 μg/L,respectively.After iodine supplementation,the medians UI level in 2007,2008,2009 and 2010 were,respectively,pregnant women:136.0,187.8,118.2,175.8 μg/L; lactating women:135.2,159.8,187.5,163.5 μ g/L; infants:139.6,174.7,190.7,168.4 μg/L.Before iodine supplementation,the DQ value of infants was 92.8 ± 16.3,and the average score of gross motor,fine motor,adaptive capacity,language and social behavior was 93.7 ± 20.0,91.4 ± 20.0,92.4 ± 19.0,90.3 ± 20.0,96.4 ± 22.1,respectively.After iodine supplementation,the DQ value of infants was 104.3 ± 13.8,and the average score of gross motor,fine motor,adaptive capacity,language and social behavior was 104.8 ± 21.5,104.1 ± 17.2,104.8 ± 16.1,99.9 ± 19.1,108.0 ± 22.7,respectively,which were higher than that before iodine supplementation (t =-10.43,-10.77,-13.78,-14.28,-9.96,-15.33,all P ＜ 0.01).Conclusions Iodine deficiency at the critical period of brain development can affect the intellectual development of infants and young children at all functional areas of intelligence.Iodine supplementation at the critical period of brain development can prevent mental retardation caused by iodine deficiency disorders.

Objective To evaluate the outcome of primary total knee arthroplasty (TKA) in treatment of stiff knee, and discuss the key points of operative technique and rehabilitation. Methods From February 2005 to April 2009, 23 patients with 34 stiff knees were treated with primary TKA. The study included 3 males (4 knees) and 20 females (30 knees), with the mean age of 56.9 years. Primary disease of the patients included rheumatoid arthritis (26 knees in 15 cases) and osteoarthritis (8 knees in 8 cases). Varus deformity was found in 10 knees (5°-15°), and valgus was found in 5 knees (5°-10°). Evaluations included preoperative and postoperative range of motion (ROM) measurement, hospital for special surgery knee score (HSS), blood loss, operative time and assessment of postoperative complications. Results All patients were followed up. The mean follow-up time was 32.2 (range, 24 to 40) months. At the final follow-up visit, the HSS score increased from 42.9±5.2 preoperatively to 85.7±4.3, the range of motion increased from 42.6°±5.7° preoperatively to 89.2°±10.5°. Sixteen knees in 12 cases underwent manipulation at 3 to 8 weeks postoperatively for unsatisfied ROM, but ROM was still less than 90° in 8 knees at the last follow-up. The average blood loss were (632.4±180.2) ml in first 24 hours (450-850 ml) and the operative time were (98.1±18.6) min (80-150 min). Deep venous thrombosis was found in 3 patients. All the symptoms relieved after anticoagulant therapy. Postoperative varus deformity was seen in one patient, but the function of knee was good. No revision was needed. Conclusion Primary total knee arthroplasty is reliable method in treatment of stiff knees. Sufficient soft tissue release during the operation, postoperative muscle strength exercise and essential manipulation are key points for satisfactory outcomes.

China ; Heart Transplantation ; methods ; Humans

Endoplasmic reticulum stress (ERS) is an adaptive process in response to circumstantial changes, but excessive and/or prolonged ERS can induce cell apoptosis. C/EBP homologous protein (CHOP) is a very important marker participating in ERS-associated cell apoptosis, while the role of the myocyte apoptosis induced by CHOP remains unclear in the development of hypertrophy. The present study aimed to investigate the effect of CHOP-mediated ERS-associated apoptosis on myocardial hypertrophy induced by abdominal aortic constriction in rats. Healthy male Wistar rats were randomly divided into model group (n=45) and control group (n=40). The rats in model group received abdominal aortic constriction. Hemodynamic changes, whole heart weight/body weight (HW/BW) and left ventricular weight/body weight (LVW/BW) were measured on 1 d, 3 d, 7 d, 14 d and 28 d after surgery, respectively. The mRNA expression of glucose-regulated protein 78 (GRP78), calreticulin (CRT) and CHOP, which are important markers of ERS, were detected by RT-PCR, and Western blot was used to assess the protein level of GRP78, CRT, CHOP, and apoptosis-associated proteins, Bax and Bcl-2. The results obtained were as follows. Compared with control group, the blood pressure, LVW/BW, and HW/BW of rats in model group increased significantly and cardiac function enhanced compensatively on 7 d after surgery, and increased progressively during the experiment. As early as 1 d after surgery, the mRNA level of CRT in model group increased by 136% (P< 0.01) compared with control, while the protein expression increased by 69.2% on 7 d after surgery (P<0.01). Both mRNA and protein expression of GRP78 increased by 20% and 186% (P<0.01) respectively on 7 d after surgery, and the expression sustained high level during the experiment afterwards. Correlation analysis indicated a positive correlation between +dp/dt(max) and CRT protein expression (r=0.780, P<0.01) as well as GRP78 protein expression (r=0.694, P<0.01). Prolonged ERS triggered myocyte apoptosis, as both the mRNA and protein level of CHOP in model group increased by 22.2% (P<0.01) and 76.0% (P<0.01) respectively compared with control on 7 d after hypertrophy (14 d after surgery), and meanwhile, the protein expression of pro-apoptotic Bax increased by 41.1% (P<0.01) and anti-apoptotic Bcl-2 protein expression decreased by 25.5% (P<0.01). Correlation analysis indicated a positive correlation between CHOP and Bax expression (r=0.654, P<0.01), and a negative correlation between CHOP and Bcl-2 expression (r=-0.671, P<0.01). These results suggest that abdominal aortic constriction induces a significant up-regulation in ER molecular chaperones at early stage of post-surgery, indicating that ERS response is activated in the rat heart; while prolonged ERS could lead to myocyte apoptosis, and CHOP-mediated ERS-associated apoptosis may contribute to myocardial hypertrophy. We speculate that cell apoptosis may take part in the regulation of myocardial hypertrophy and heart failure, and determine the progression of decompensated hypertrophy.
Animals ; Aorta ; physiopathology ; Apoptosis ; Calreticulin ; metabolism ; Constriction ; Endoplasmic Reticulum Stress ; Heat-Shock Proteins ; metabolism ; Hypertrophy ; pathology ; Male ; Myocardium ; pathology ; Rats ; Rats, Wistar ; Transcription Factor CHOP ; metabolism ; Up-Regulation ; bcl-2-Associated X Protein ; metabolism