Objective To observe the effect of small interfering RNA(siRNA)expression plasmids targeting transforming growth factor p receptor(TαR)Ⅰ gene on the collagen synthesis of hepatic stellate cells(HSCs).Methods Three siRNA expression plasmids were designed and constructed according to TBR Ⅰ sequence.Then the plasmids were transfected into HSC-T6 using 1ipofectamine2000 reagent. The mRNA and protein expressions of TβR Ⅰ were analyzed by reverse transcription polymerase chain reaction(RT-PCR)and Western blot technique, respectively. The cell proliferation was detected using methylthiazo-lyldiphenyl-tetrazolium bromide(MTT)methods. Concentrations of haluronic acid and type Ⅲ pro-collagen in the supernatants were determined by radioimmunoassay. The data were analyzed using least significant difference(LSD).Results Three recombinant plasmids expressing siRNAs were successfully constructed and confirmed by restriction enzyme assay. Compared with the blank control,all the three recombinant plasmids could inhibit the expressions of TβR Ⅰ mRNA,of which plasmid expressing siRNA2 exhibited the strongest inhibitory effect(psiRNA1 group:t=7.354,P＜0.01;psiRNA2 group:t=9.214,P＜0.01;psiRNA3 group:t=5.967,P＜0.01).The expressions of TβR Ⅰ protein were also reduced by all the three recombinant plasmids,of which the plasmid expressing siRNA2 showed the strongest inhibitory effect(psiRNA1 group: t=6.324,P＜0.01;psiRNA2 group:t=8.741,P＜0.01;psiRNA3 group:t=4.128,P＜0.01).The proliferation activity and collagen synthesis of HSCs also decreased in all three HSC groups treated with recombinant plasmids, of which, again, plasmid expressing siRNA2 exhibited the strongest inhibitory effect. However, no significant change was observed in HSCs transfected with non-related siRNA. Conclusion Recombinant plasmids targeting TβR I can inhibit collagen synthesis, which suggests a novel target for gene therapy of liver fibrosis.

Objective To investigate the effect of different doses of ketamine on inflammatory cytokines in rabbits with severe burn at early stage and preliminarily approach its regulatory action on early stage of inflammatory reaction due to stress of trauma.Methods Forty healthy male New Zealand rabbits were randomly divided into four groups in accord with the random number table method: normal control group, scald model group, ketamine analgesia group and ketamine anesthesia group. Before scald, pentobarbital sodium was used for anesthesia, afterwards catheters were inserted into internal jugular vein and internal carotid artery respectively ready for use, and 24 hours later, Ⅲ degree scald at the animal back and buttocks occupying 30% total body surface area (TBSA) was performed as the scald model for all the rabbits except those in normal control group. In ketamine analgesia group, after scald for 0.5 hour, 0.5 mg/kg ketamine intravenous injection was given to the rabbits as the loading dosage and then persistent intravenous pump infusion of 9μg·kg-1·min-1 ketamine was applied for all together 24 hours. In ketamine anesthesia group, after scald for 0.5 hour, 1.5 mg/kg ketamine intravenous injection was given to the rabbits, and then persistent intravenous pump infusion of 45μg·kg-1·min-1 ketamine was applied for 4 hours to maintain systemic anesthesia. In normal control and scald model groups, only intravenous infusion of equal amount of normal saline was given to the rabbits. The amount of intravenous transfusion in each group and the total dosages of ketamine used in ketamine analgesia group and ketamine anesthesia group were recorded. Before scald and 0.5, 6, 12, 24 hours after scald, arterial blood gas analyses were made, and the levels of serum interleukins (IL-1, IL-6) and tumor necrosis factor-α (TNF-α) were determined.Results Although the indexes of blood gas analysis were changed in the four groups, they were all in the normal range, showing that the respiratory function was in the normal range and indirectly reflecting that the circulatory function was also in the normal range, thus the effects on cytokines by factors of respiratory and circulatory functions were ruled out. The levels of IL-1, IL-6 and TNF-α before scald showed no statistically significant differencesamong the four groups (allP > 0.05). From 0.5 hour after scald, the levels of IL-1, IL-6 and TNF-α were markedly higher in model group than those of normal control group [IL-1 (ng/L): 30.27±0.93 vs. 13.79±1.11, IL-6 (ng/L): 47.22±1.49 vs. 46.31±4.12, TNF-α (ng/L): 243.39±20.85 vs. 190.95±14.97, allP < 0.05], and the situation continued until 24 hours after scald; the levels of IL-1, IL-6 and TNF-α from 6 hours after scald were significantly decreased in ketamine analgesia and ketamine anesthesia groups compared with those in the model group, and from 12 hours after scald, the degrees of descent in levels of the above indexes in ketamine analgesia group were more obvious than those in ketamine anesthesia group [IL-1 (ng/L): 19.28±2.51 vs. 40.12±10.31, IL-6 (ng/L): 52.10±4.23 vs. 72.20±10.11, TNF-α (ng/L): 246.03±20.74 vs. 313.71±27.34, allP < 0.05].Conclusion The low-dose ketamine analgesia and ketamine anesthesia have certain degree of inhibitory effect on the expression and release of inflammatory cytokines at the early stage in rabbits with severe burn, the effect of long-term low-dose ketamine analgesia being more significant.

Objective To study prevention and treatment for patients during the intermittent period of maintaining hemodialysis complicated by acute left heart failure. Methods The treating process of 126 examples suffered from acute left heart failure during the intermittent period of 14 225 person-time maintaining hemodialysis in 143 patients were retrospectively analyzed. Results In this case group, 0.89% suffered from acute left heart failure during the intermittent period of hemodialysis and the successful rescuing rate was 88.9%. Conclusions Applying the treatment of emergency hemodialysis, quickly filtering body fluid, the curative effect is reliable and rapid. It is one of the best ways to cure patients suffered from acute left heart failure during the intermittent period of maintaining hemodialysis. Combined with other treatments, it will be able to improve the successful rate, and ensure the sufficiency of hemodialysis. United with other steps of reducing blood pressure, such as using angiotensin converting enzyme inhibitors, angiotensin II receptor blocker, and ? receptor blocker, it is an effective countermeasure to prevent patients suffering from acute left heart failure during the intermittent period of maintaining hemodialysis, and effectively reduces its appearance as well.

Objective To observe the dynamic expressions of kielin/chordin-like protein (kcp) in mouse model of hepatic fibrosis and the effects of bone morphogenetic protein-7 (BMP-7)intervention on the expressions,and to explore a new target for treatment of fibrosis.Methods A total of 50 healthy male ICR mice were divided into three groups:control group(n=10) ; model group (n=30) and BMP-7 treatment group (n=10).The model group was further divided into three subgroups according to different time points:subgroups of 4,8 and 12 weeks with 10 mice in each subgroup.The mouse model of hepatic fibrosis was established by hypodermic injection of carbon tetrachloride(CCl4 ).The mice in BMP-7 treatment group began to receive human recombmant BMP- 7via intraperitoneal injection after 8 weeks of the first administration of CC14 and lasted for 4 weeks.The serum levels of alanine aminotransferase (ALT),aspartate transaminase (AST) and albumin (Alb) were detected.The pathological changes of liver were observed under optical microscope after HE and Masson staining.The dynamic expressions of kcp mRNA and protein of each group were detected by reverse transcription-polymerase chain reaction,immunohistochemistry and Western blot.The comparison of means among groups was done by univariate ANOVA.Results In model group,ALT and AST levels increased,while Alb level gradually decreased,and peaked at week 12.BMP-7treatment could reduce the changes,and there were significant differences among groups (F=23.501,34.600 and 16.244,respectively; all P＜0.05).In normal control group,the expressions of kcp mRNA and protein were low,while those in model group were gradually increased.BMP-7 treatment could achieve remission and the changes were all significantly different among groups (F=30.362 and 10.727,respectively; P＜0.01 or 0.05).The expression of kcp mRNA was positively correlated with levels of transforming growth factor (TGF)-β1 mRNA and BMP-7 mRNA (r=0.760 and 0.769,respectively; both P＜0.05).Conclusions BMP 7 can improve the hepatic fibrosis in mice.kcp may play an important role in the occurrence and development of liver fibrosis which is a potential therapeutic target for hepatic fibrosis.

Objective To set up a system in vitro to rapidly recover plasma proteins lost during artificial-liver treatment.Methods The polyprotein precipitation was obtained by all proteins whose isoelectric point pH value were between 7.3 and 5.1,which collided with each other and aggregated using gradient pH co-precipitation(adding 1 mol/L citric acid slowly in the plasma solution to change the pH values gradually from 7.3 to 5.1 in 5 h)combined with salting out(degree of saturation of NaCl is 33%,reacted for 5.5 h at 4℃)or low-temperature ethanol precipitation(40% ethanol, reacted for 5.5 h at -7℃)so that to get rid of toxicants by discarding the supernatant.Results In the range of pH 5.1-7.3,50%(29g/57g)of the total plasma proteins had been recovered by the gradient pH salting out and 41%(25 g/61g)by the gradient pH low-temperature ethanol co-precipi- tation.The protein remained in the supernatant was mostly albumin and its combined bilirubin.The levels of total bilirubin decreased to 0.07% and 0.06% of the original levels by these two methods respectively and the serum HBV DNA level decreased to be undetected(quantitative PCR).Conclu- sions The proteins with close isoelectric point can co-precipitated with the presence of high concen- tration of NaCl or low-temperature ethanol and by changing the pH value gradually.The total protein in the discarded plasma during artificial-liver treatment can be recovered rapidly using the gradient pH coprecipitation.

Objective To explore the dynamic expressions and the significance of Notch/Jagged signal pathway in rat model of hepatic fibrosis. Methods A total of 42 healthy male SD rats were randomly divided into normal control group (n=6) and model group (n= 36). The model group was further divided into six subgroup according to different time points: subgroups of 4 days, 1, 2, 4, 6 and 8 weeks with six rats in each subgroup. The rat model of hepatic fibrosis was induced by dimethylnitrosamine (DMN). The serum levels of alanine aminotransferase (ALT), aspertate aminotransferase (AST), albumin (Alb) and hyaluronic acid (HA) were detected dynamically after 4 days, 1,2,4,6 and 8 weeks of injection. The liver tissues were observed under optical microscope after HE and Masson staining. Notch-1, Jagged-1 mRNA and protein in liver were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. The comparison of means among groups was done by univariate ANOVA. Results The hepatic fibrosis model was successfully induced by DMN injection and pseudolobules were found after 4 weeks of injection. The serum levels of ALT, AST, Alb and HA were all increased after 4 day of injection and peaked at week 4 which were all significantly higher than those in control group (F=83.10, 104.63, 54.24, 203.81,respectively; all P＜0.05). The expressions of Notch-1, Jagged-1 mRNA and protein in model group were all significantly increased than those in control group (F=282. 44, 369.14, 374.17, 256. 14,respectively;P＜0. 01). And the expressions of Notch-1, Jagged-1 were closely correlated with the hepatic fibrosis stages and transforming growth factor β1 (TGFβ1) expression (r=0. 821, 0. 917,0. 767,0. 844, respectively; P＜0. 01 ). Conclusions The Notch/Jagged pathway may participate in the development of hepatic fibrosis, which is closely correlated with the progression and severity of liver fibrosis.

Objective To explore the normal range of the caudate nucleus' volume in Chinese adults of the Han nationality and provide morphological data for the construction of database for Chinese Standard Brain.Methods This was a clinical multi-center study.One thousand Chinese healthy volunteers (age range=18 to 70)recruited from 16 hospitals were divided into 5 groups,i.e.,Group A(age range=18 to 30),B(age range=31 to 40),C(age range=41 to 50),D(age range=51 to 60),and E(age range=61 to 70).Each group contained 100 males and 100 females.All of the volunteers were scanned by MR using T1 weighted three-dimensional magnetization prepared rapid acquisition gradient echo sequence.The volume of eaudate was measured manually using 3D volume analysis software.The difierence of volumes of the eaudate between male and female were analyzed by independent sample t-test,and among age groups by ANOVA.Pearson's correlation coefficient was used to characterize the relationship between volunles and age.The differences of measurements between left and right caudate nucleus were analyzed by paired t test.Results (1)rnle mean volume of bilateral caudate nucleus in healthy Chinese adults wag(10.973±1.647)cm3.The mean volume of the the male's left and right caudate nucleus were(5.656±0.860)and (5.671±0.855)cm3 respectively,no significant differences were found between the volume of left and right eaudate nucleus(t=1.230.P>0.05).The mean volume of the the female's left and right eaudate nucleus were(5.287±0.774)and(5.331±0.766)cm3 respectively,and the right's wag larger than the left's with significant differences(t=3.999,P<0.01);(2)Pearson correlation analysis showed a significant negative correlation between the nucleus volume and age(male and female's,left and fight)(r=-0.561,-0.568,-0.548,-0.552,P<0.05).Conclusion With high-resolution MRI and 3D volumetric analytic software(Midob),the volume of the caudate nucleus can be accurately measured,so as to provide the basic data for establishing Chinese adults' standard digital brain.and provide nomad control standards for caudate nucleus volume-related diseases.

Objective To explore the normal range of cingulate cortex volumes of Chinese adults of the Han nationality and its relationship with age, which provide morphological data for the construction of database for Chinese Standard Brain.Methods This is a clinical multi-center study.One thousand Chinese healthy volunteers ( age range = 18 to 70) recruited from 15 hospitals were divided into 5 groups, i.e.,Group A (age range = 18 to 30), B (age range=31 to 40), C (age range =41 to 50), D (age range =51 to 60), and E (age range =61 to 70).Each group contained 100 males and 100 females.All of the volunteers were scanned by MR using T1 weighted three-dimensional magnetization prepared rapid acquisition gradient echo sequence.Cingulate cortex volume (including bulk volume and the left/right volume) was measured semi-manually using 3D volume analysis software.Cingulate cortex volumes among age groups were compared by one-way ANOVA.Right and left cingnlate cortex volumes between sexualities were analyzed by paired samples t test.The relationship between cingulate cortex volume and age was analyzed by Pearson correlations and regression analysis.Results Cingulate cortex volumes of male and female were (20 347 ± 2504) and ( 19 432 ± 2184) mm3 respectively, and the male's was significantly larger than that of female's (two sample t'-test for independent samples, t'= 6.156, P < 0.05 ).Right and left cingnlate cortex volumes of male were ( 10 717 ± 1629) and (9630 ± 1498) mm3 respectively, and those of female's were ( 10 064 ± 1407 ) and ( 9368 ± 1441 )mm3 respectively.The volumes of cingulate cortex were significantly different between right and left in male or female ( t = - 12.960, - 8.511, P < 0.05 ),and right was larger than left.Bilateral cingulate cortex volume in male among group A, B, C, D and E[left: ( 10 132 ± 1291 ), ( 10 113 ± 1638), (9599 ± 1576), (9594 ± 1288), (8710 ± 1212) mm3 ; right:(11 212±1442), (11 096±1602), (11 040±1403), (10 633±1638), (9604±1522) mm3] had statistical differences (F = 16.738, 18.707, P < 0.01 ) ; and those in female among five age groups[left:(9689 ± 1426), (9652 ± 1676), (9347 ± 1500), (9098 ± 1225), (9053 ± 1233) mm3 ; fight: ( 10 558 ±1325), ( 10 266 ± 1463), ( 10 100 ± 1497), (9779 ± 1304), (9617 ± 1254) mm3] also had significant differences (F = 16.859,7.528,P <0.01 ).Bilateral cingnlate cortex volume in both male and female were negatively correlated with age ( r = - 0.330, - 0.324, - 0.169, - 0.243, P < 0.05 ), though the correlation coefficient is not high.Conclusions Cingnlate cortex volume could be accurately measured on the high-resolution MRI with 3D volume analysis software, which can provide morphological data for the construction of database for Chinese Standard Brain.The results may provide normal range for the diagnosis of the volumetric deficits of cingulate cortex.

Objective To explore the normal range of the volume of frontal lobe in Chinese adults of the Han nationality and provide morphological data for the construction of database for Chinese Standard Brain. Methods This is a clinical multi-center study. Two hundred Chinese healthy volunteers (age range= 18 to 70) recruited from 16 hospitals were divided into 5 groups, i. e. , age range from 18 to 30, age range from 31 to 40, age range from 41 to 50, age range from 51 to 60, and age range from 61 to 70. Each group contained 20 males and 20 females. All of the volunteers were scanned by MR using T1 weighted threedimensional magnetization prepared rapid acquisition gradient echo sequence. We used the manual method to trace the region of interest and measured the left and right frontal lobe volumes separately. All the data were analyzed with SPSS (version 13.0). The sex differences in the frontal lobe volumes were analyzed by independent-samples t test, and the side differences were analyzed by paired-samples t test. Correlation and regression analysis was used between the age and the frontal lobe volumes. Results In 200 healthy Chinese Han volunteers, the total frontal lobe volumes was (563 ± 73) cm3. For male, the volumes of the left and the right frontal lobe were (288±42) cm3 and (292 ±41 ) cm3 ,respectively. The volumes of the left and right frontal lobe in 100 women were (273 ±30) cm3 and (274 ± 30) cm3 respectively. The differences of sex (t = 3. 334, P < 0. 05 ) and side ( t = - 3.09, P < 0. 05 ) in the total frontal lobe volumes reach significant. There was no significant differences in women ( t = - 1. 304, P > 0. 05 ). There were negative correlations between the frontal lobe volumes and age in men and women ( r = - 0. 586, - 0. 498, P <0. 01 ). Conclusions The total frontal lobe volume of men was larger than that of women. The volume of the right frontal lobe was larger than the left frontal lobe in men, and the asymmetries didn't exist in women.The total frontal lobe volumes were both shrinking with age in men and women, which was more rapid in men than in women.

Objective To measure the volume of hippoeampal formation(HPF) in healthy Chinese Han adults and provide database for researching on a variety of diseases associated with alteration of hippocampal structure. Methods This is a clinical multi-center study. One thousand Chinese healthy volunteers (age range = 18 to 70) recruited from 15 hospitals were divided into 5 groups, i. e. , Group A (age range = 18 to 30), B (age range =31 to40), C (age range =41 to 50), D (age range=51 to 60),and E (age range =61 to 70). Each group contained 100 males and 100 females. All of the volunteers were scanned by MR using T1 weighted three-dimensional magnetization prepared rapid acquisition gradient echo sequence. The margin of HPF were outlined manually for each side. Using multiple linear regression,relationships between hippocampal volume and sex, age, weight and height were analyzed respectively.Independent two sample t test was used to study the differences between male and female and between lcft and right. The differences of hippoeampal volume among age groups were analyzed by ANOVA. Results Hippocampal volume for left and right side were (4752 ± 659) and (5032 ± 660) mm3 respectively. The volume of HPF is significant correlated with gender and age, but without relevance to height and weight ( left and right r = 0. 283,0. 311, F = 30. 127,37. 050 ,P < 0. 01 ). The volumes of left and right hippocampus were (4897 ± 670), (5192 ± 667)mm3 respectively for men, and (4647 ± 624) and (4904 ± 630) mm3 for women. The right hippocampal volume was larger than the left ( t = 7. 030,6. 696, P < 0. 05 ). On both sides, men have larger hippocampus than that of women ( t = 6. 586,7. 326, P < 0. 01 ). The volumes of the left hippocampus among the five age groups in the male were (4981 ± 684), ( 5003 ± 609 ), (4976 ± 657 ),(4841 ± 631 ), (4631 ± 681 ) mm3 respectively, while the volumes of the right hippoeampus were (5340 ±647) ,(5276 ±582), (5264 ±620), (5133 ±661 ), (4894 ±699) mm3 respectively. Among age groups,the differences were statistically significant ( left and right F = 5. 737,7. 607, P < 0. 01 ). For male, there was significant difference of bilateral hippoeampus between the 18-30, 31-40, 41-50 years group and 61-70 years group ( P < 0. 05 ) and there was no significant difference among the other groups ( P > 0. 05 ).There was no significant difference of hippocampal among different groups in women (P > 0. 05 ).Conclusions With high-resolution MRI, the volume of the HPF was accurately measured, so as to provide the basic data for research of the hippocampus-related disease.