Objective:To determine phenolphthalein illegally added into diet health products by Raman spectroscopy. Methods:Raman spectroscopy was used to determine phenolphthalein added into diet health products. The diet health products were extracted by suitable solvents, and then the extracting solution was measured by Raman spectroscopy. Results: A calibration curve was built and the analysis was performed on the samples with phenolphthalein at different concentrations. The results were accordance with the real added amount. Conclusion:With simple sample preprocessing method, Raman spectroscopy can be used in the fast detection of phe-nolphthalein illegally added into diet health products. The method is fast and simple with low cost, and can provide both qualitative and quantitative results. The detection limit is 1% for phenolphthalein in diet health products.

Objective To explore the stress response and immune balance in patients with severe trauma hemorrhage after fluid resuscitation and to clarify the clinical effect of Propofol administered with continuous intravenous infusion pump on it.Methods With prospective,randomized and control analysis,54 patients treated with fluid resuscitation following severe trauma hemorrhage admitted from October 1st,2008 to December 1st,2009 were studied.Another 20 healthy volunteers were enrolled as control group (C group). Patients were randomly divided into:conventional treatment group (R group,n =27 ) and conventional therapy combined with propofol treatment ( P group,n =27) as per gender,age,ISS score,estimated blood loss four factors and the principle ofminimum distribution imbalance index.HR,MAP,the levels of stress hormones [ norepinephrine (NE),cortisol (Cor) ],immune function ( T lymphocyte subsets Th1/Th2) and other biomarkers were observed.Mortality within 28 d between R group and P group was compared.ANOVA was used for the comparison of biomarkers,and independent samples t test was employed to compare variables between the two groups. Results Plasma cortisol (Cor),norepinephrine (NE),alanine aminotransferase (ALT),creatinine ( Cr),blood glucose,and Th1 and Th2 lymphocytes in patients were significantly higher than those in healthy volunteers of group C (P ＜0.01 ),and Th1/Th2 was significantly lower than those in healthy volunteers of C group (P =0.001 ).The ALT (48 h),Glu (6 h),NE (24 h and 48 h) and Cor (6 h and 24 h) of patients in P group were lower than those in R group ( P ＜0.05 ),and Th1 and Th2 were lower than those in R group at different intervals ( Th1:24h P =0.028,48 hP=0.002 ; Th2:6 h P=0.033,24 h P=0.007,48 h P=0.009),and Th1/Th2 ratio (48 h) was significantly higher in P group than that in R group (P =0.028),but there was no significant difference in mortality within 28d between the two groups. Conclusions Strong stress response,immune suppression and organ dysfunction can occur in the early stages of severe trauma hemorrhage after fluid resuscitation.Propofol can inhibit excessive stress response,modulate the immune balance and protect the important organs in those exsanguination patients from severe trauma.

Objective To discuss the treatment of renal or adrenal tumor with cancer thrombus in the inferior vena cava. Methods From Jan 1984 to Apr 2008,29 cases of renal or adrenal malignancy with thrombosis involving the inferior vena cava underwent treatment.The diagnoses were confirmed by Doppler uhrasonography,CT and MRI.In the 29 surgical patiens the tumor thrombus was level I in 7,level Ⅱ in 10,level Ⅲ in 8 and levelⅣin 4.According to TNM classification,23 cases were classified to T2N.M.,1 case was T2Nl Mo,1 case was TzNlMl,1 case was T3NoMo,2 case were T3NlMl and 1 case was T3N2Mo.The mean tumor size was 8.7(4.O-16.O)cm in diameter.The mean tumor thrombosis length was 3.2(2.5-4.0)cm in level I,5.3(4.5-6.0)cm in level Ⅱ,8.2(6.5-9.O)cm in levelⅢand 15.1(12.0-18.5)cm in level IV. Results The operation was performed succesgfullv in 29 patients Patholocieal examination showed that 18 cases of clear cellcarcinoma,3 cases sarcomatoid carcinoma,2 cases renal papillary adenocarcinoma,1 case renal cell carcinoma (undifferentiated),1 case granule carcinoma,3 cases adrenocortical carcinoma and 1 case metastatic malignant melanoma of adrenal gland.Of 29 patients,3 were out of contact.Twenty-six patients were followed up for 35(0-62)months after treatment,3-and 5-year survival rates were 15/26 and 11/26.Three-year survival rates for stage T2 and T3 were 14/22 and 1/4.Five-year survival rates for stage T2 and T3 were 10/22 and 1/4.Three-year survival rates for level I、Ⅱ、Ⅲ andⅣ were 4/6,5/8,5/8 and 1/4.Five-year survival rates for level I,Ⅱ、Ⅲ andⅣ were 3/6,4/8,3/8 and 1/4.Three-year survival rates for a tumor thrombus in the below or above diaphragm were 14/22 versus 1/4,5-year survival rates were 10/22 versus 1/4.Three-year and 5-year survival rates for the patients without distant metastases and lymph node involvement were 12/18 and 9/18.Three-year and 5-year surviral rates for the patients with distant metastases and lymph node involvement were 3/8 and 2/8.The 3 surgical patients with metastatic disease died at 6,10,22 months. Conclusions Surgical treatment could be the preferred approach for the patients of renal or adrenal tumor with cancer thrombus in the inferior vena cava without distant metastases and lymph node involvement.It could improve the quality of life and may prolong survival.

Objective To discuss the clinical feature ,diagnosis and treatment of the occult cerebrospinal fluid rhinorrhea after tracheotomy in patients with severe traumatic brain injury .Methods The clinical data of 18 cases of the occult cerebrospinal fluid rhinorrhea after tracheotomy in patients with severe traumatic brain injury were retro -spectively analyzed .Results 15 cases showed involuntary swallowing movements ,frequent stimulus-likecough, abnormal increased secretions in the oral and nasal;3 cases performance of aspiration ,hypoxemia ,respiratory distress . After a three -dimensional thin skull CT , cisternography , nasal endoscopic examination can confirm the diagnosis . After the treatment with replacing the tracheostomy tube with a balloon ,continuous lumbar drainage ,endoscopic repair leak,the cerebrospinal fluid rhinorrhea were cured .Conclusion Patients with occult cerebrospinal fluid rhinorrhea performance the diversity and easily missed ,early detection and timely treatment can prevent cerebrospinal fluid rhi-norrhea delayed healing and intracranial infection and promote patient recovery .

Objective To investigate the impact of hyper-homocystinemia (Hcy)on cardiac remolding in spontaneously hypertensive rat(SHR)and the protective effects of folic and vitamin B12 on the remolding.Methods The seven-week-old SHR were divided into four groups according to different diet for 20 weeks.The control group was fed a perfect compound diet without methionine,the methionine group was fed a perfect compound diet added with 2 ％ methionine,the treatment group was fed as that of methionine group,but added with vitamin B12 0.09 mg/(kg.d)and folate 4 mg/(kg.d)in the last 12 weeks,the mixture group was fed as that of methionine group in the first 8 weeks,and as that of control group.At the end of the intervention period,plasma levels of Hcy and matrix metalloprotease(MMP)-9 were measured.The blood pressure was detected by the non-invasive blood pressure sensing device.The cardiac structure and function were detected using echocardiography and invasive hemodynamic method.Picro-sirius red(PSR)staining was used to check the fibrosis in paraffin embedding ventricle section.Results Plasma levels of Hcy and MMP-9 were increased in the methionine group,while there was no significant difference in plasma levels of Hcy and MMP-9 among the treatment group,the mixture diet group and control group.Rats in methionine group had the largest heart weight among four groups,and systolic blood pressure was lower in methionine group than in the control group(P＜0.05).Compared with the other three groups,the methionine group showed that the left ventricular end-diastolic pressure(LVEDP)was increased and-dp/dt maxwas decreased.Meanwhile,LVEDP and-dp/dt m,x had no significant difference between the control group,treatment group and mixture group.The methionine group showed the most severe decrease in the left ventricular fractional shortening(LVFS)and +dp/dt While LVFS and + dp/dt max had no statistical significance between the treatment group and control groups,but the mixture group had lower LVFS and +dp/dt max values as compared with the control group(both P＜0.05).Perivascular fibrosis was increased significantly in methionine group as compared with the other three groups,while the treatment and control groups had similar perivascular fibrosis,but the mixture group had higher perivascular fibrosis than did the control group,and had similar perivascular fibrosis to the treatment group.Fibrosis deposition in myocardium mesenchymal was similar to those observed in perivascular fibrosis,but the treatment group showed a more improvement of collagen deposition as compared with the mixture group.Conclusions Hyper-homocystinemia can accelerate systolic and diastolic dysfunction through accumulating fibrosis in myocardium and perivascular wall which lead to adverse cardiac remolding.Folate and vitamin B12 have protective effects on heart function by reversing the adverse changes through decreasing Hcy level.

Objective To report our self-designed guide module for placement of posterior column lag screws in anterior-posterior column acetabular plate using CT reconstruction data.Methods The CT scan data of 50 normal adult pelves were collected from February 2012 to April 2013,involving 30 males and 20 females with an average age of 46.4 years(range,from 25 to 69 years).The data were imported into Mimics 10.01 software for reconstruction of semi-pelvic models.Virtual cylindrical implants were placed intraosseously in both the left and the right posterior columns.The perpendicular distance (OP) from the insertion point O of the virtual cylindrical implant to the arcuate margin (P) and the distance (PI) from the point P to the point I,the crosspoint of the extension line of the ischial ramus and the arcuate margin were measured respectively.The angle (∠φ) between the direction of screws and the plane of guide module and the angle (∠θ) between the direction of screws and the long axis of guide module were also measured respectively.Results The average length of PI was 0.98 ± 0.13 cm,with 1.08 ± 0.22 cm in females and 0.95 ± 0.27 cm in males.The difference between genders was not statistically significant (P ＞ 0.05).The average length of OP was 1.09 ± 0.26 cm,with 1.06 ± 0.29 cm in females and 1.12 ± 0.24 cm in males.The gender difference was not statistically significant either (P ＞ 0.05).The mean value of ∠ φ was 55.43° ± 3.64°,with 55.33° ± 4.00° in females and 55.50° ±3.43° in males.The difference between genders was not statistically significant (P ＞ 0.05).The ∠θ value in females was 39.21 ° ± 2.45° and 35.58° ± 2.31 ° in males.The gender difference was statistically significant (P ＜ 0.05).Conclusions In design of the guide module,the nail holes should be located about 1 cm away from both the posterior edge and the medial edge,the angle between the screw direction and the module plane should be approximately 39° in females and 35° in males,and the angle between the screw direction and the long axis of the module approximately 55°.

BACKGROUND:Pathogenesis of Parkinson’s disease is not completely understood, and there is yet no effective therapy that can prevent the neurodegenerative process of the disease fundamentaly. OBJECTIVE:To explore the effects of pituitary adenylate cyclase-activating polypeptide (PACAP) on lactacystin-induced Parkinson’s disease dopaminergic PC12 cel apoptosis and its molecular mechanism. METHODS: Under induction by nerve growth factors, PC12 cels differentiated into dopaminergic neurons, and then were treated with different concentrations of lactacystin for different time. When the cel survival rate was about 50%,the concentration and action time oflactacystin were selected to establish cel models of Parkinson’s disease. In the study, there were control group, lactacystin group, PACAP1-27 group (intervention group 1) and PACAP1-27+PACAP6-27 co-intervention group (intervention group 2). Changes of cel morphology were observed under inverted microscope; cel viability was detected with MTT method; the expression of endoplasmic reticulum stress specific protein caspase-12 was detected by western blot. Then the action of PACAP1-27 and PACAP6-27 to the cytoxicity of lactacystin was observed. RESULTS AND CONCLUSION: With different concentrations and action time of lactacystin, the viability of PC12 cels presented a concentration- and time-dependent decline. When the lactacystin at 20μmol/L acted for 24 hours, the cel viability was declined by about 50%. Under same conditions of lactacystin concentration and action time (20 μmol/L, 24 hours), the cels in the lactacystin group appeared to have damaged changes, declined cel viability, and increased caspase-12 activity in comparison with the control group (P< 0.01). Compared with the lactacystin group, the cel damage was relieved and cel viability was increased significantly in the intervention group 1 as wel as the expression of caspase-12 was decreased (P < 0.01). Experimental findings in the intervention group 2 were similar to those in the lactacystin group. These results suggest that lactacystin, an ubiquitin proteasome inhibitor, can lead to cel damage; PACAP1-27 plays a protective role by regulating the above-mentioned signal pathway. As one PACAP1-27 receptor antagonist, PACAP6-27 can attenuate this effect of PACAP1-27.

ObjectiveTo observe whether lipopolysaccharide (LPS) derived fromEscherichia coli (E.coli) can induce apoptosis of murine platelets in vitro.Methods Washed platelet suspension was prepared and adjusted to the final concentration of 3×108/mL. According to the difference in stimulants, samples were divided into control group (non-calcium Tyrode buffer), thrombin-treated group (1 U/mL final concentration and non-calcium TB) and LPS in different concentrations treated groups (1, 10 and 100μg/mL final concentration respectively and non-calcium TB). To each specimental group corresponding stimulus was added and incubated 30 minutes at room temperature. Chemiluminescence was adopted to determine the concentration of adenosine triphosphate (ATP) and the activity of cysteinyl aspartate specific proteinase-3 (caspase-3). The percentage of Annexin V positive platelets was determined by flow cytometry to reflect the level of phosphatidylserine (PS) exposure. Mean channel fluorescence (MCF) of platelets was determined by flow cytometry for reflecting the level of mitochondrial inner transmembrane potential (ΔΨm) depolarization.Results Compared with control group, the ATP concentration in thrombin-treated group was decreased obviously [relative light unit (RLU): (5.46±0.14)×105 vs. (6.25±0.26)×105,P< 0.05], Annexin V positive ratio [(50.43±2.45)% vs. (1.58±0.25)%,P< 0.05] and caspase-3 activity [RLU: (26.92±1.60)×103 vs. (1.30±0.10) ×103,P< 0.05] were increased obviously, and platelets MCF was lowered significantly [(8.32±0.58)×104 vs. (13.05±1.10)×104,P< 0.05], suggesting an increase inΔΨm depolarization. After being treated with different concentrations of LPS, ATP concentration, Annexin V positive ratio and caspase-3 activity were increased obviously, platelet MCF was decreased obviously, suggestingΔΨm depolarization was increased in a concentration-dependent manner. Compared with control group, 1μg/mL LPS could increase Annexin V positive ratio [(10.45±1.08)% vs. (1.58±0.25)%,P< 0.05], elevate caspase-3 activity [RLU: (14.06±0.61)×103 vs. (1.30±0.10)×103,P< 0.05], and decrease MCF significantly [(9.48±0.50)×104 vs. (13.05±1.10)×104,P< 0.05]. The ATP concentration, Annexin V positive ratio and caspase-3 activity reached maximum levels after the treatment with 100μg/mL LPS, and they were higher obviously than those of the control group [ATP (RLU): (7.00±0.03)×105 vs. (6.25±0.26)×105, Annexin V positive ratio: (55.35±2.42)% vs. (1.58±0.25)%, casepase-3 (RLU): (32.00±3.75)×103 vs. (1.30± 0.10)×103, allP< 0.05], and platelets MCF reached trough levels, and they were obviously lower than those of the control group [(4.69±0.55)×104 vs. (13.05±1.10)×104,P< 0.05].ConclusionE.coli LPS can induce an increase in ATP, PS exposure,ΔΨm depolarization and activity increase of caspase-3 on mouse platelet in vitro, which indicate that LPS can induce apoptosis of platelets in a concentration-dependent manner.

ObjectiveTo explore the relationship between thrombocytopenia (TCP) induced by lipopolysaccharide (LPS) and coagulation or inflammatory response in mouse.Methods Forty-eight C57BL/6 mice were divided into control group, low-dose, and high-dose LPS treatment groups by random number table method, and each group was subdivided into 4-hour and 24-hour subgroups randomly, with 8 mice in each subgroup. 0.5 mg/kg or 50 mg/kg LPS was injected intraperitoneally in low-dose or high-does group respectively, and equal amount of normal saline was injected in control group. Blood was collected from endocanthal vein at the specified time point, platelet count (PLT) was counted, and the levels of thrombin antithrombin complex (TAT), D-dimer, fibrinogen degradation product (FDP), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were determined by enzyme linked immunosorbent assay (ELISA).Results Compared with control group, PLT (×109/L) at 4 hours and 24 hours in low-dose and high-dose LPS groups was significantly decreased (4 hours: 660.65±180.48, 568.55±117.99 vs. 1 199.13±110.54; 24 hours:505.63±218.92, 256.33±72.86 vs. 1 229.13±1 189.37, allP< 0.05), and the changes were more obvious in high-dose LPS group compared with those of the low-dose LPS group (allP< 0.05). Factorial analysis showed that the changes in PLT were related with LPS dosage and time (F1 = 135.660,P1 = 0.000;F2 = 12.120,P2 = 0.001). It was also found that there was an interactive effect of the dose of LPS and time on PLT (F = 5.580,P = 0.007). Compared with control group, TAT, TNF-α, and IL-6 at 4 hours and 24 hours in low-dose and high-dose LPS groups were significantly decreased [TAT (ng/L) at 4 hours: 1.10±0.59, 0.22±0.13 vs. 3.47±1.73; 24 hours: 1.18±0.68, 0.39±0.29 vs. 3.19±1.27;TNF-α (nmol/L) at 4 hours: 87.35±12.29, 93.70±5.25 vs. 101.59±10.96, 24 hours: 81.94±8.26, 93.23±4.71 vs. 102.84±10.56; IL-6 (ng/L) at 4 hours: 81.78±7.82, 78.59±9.06 vs. 110.88±9.66, 24 hours: 76.03±9.85, 71.34±3.69 vs. 110.88±10.35, allP< 0.05]. TAT at 4 hours and 24 hours in high-dose LPS group was further decreased, and TNF-αat 24 hours was increased as compared with those of low-dose LPS group (allP< 0.05). TAT, TNF-α and IL-6 were influenced only by different dosage of LPS (TAT:F = 42.350,P = 0.000; TNF-α:F = 14.810,P = 0.000; IL-6:F =81.910,P = 0.000), not time (TAT:F = 0.002,P = 0.967; TNF-α:F = 0.342,P = 0.562; IL-6:F = 2.973,P = 0.092). Changes in TAT was not found to be related with the dose of LPS and its time of action, or levels of TNF-α and IL-6 (TAT:F = 0.236,P = 0.791; TNF-α:F = 0.572,P = 0.569; IL-6:F = 0.774,P = 0.468). The dosage of LPS and time of admission showed no influence on D-dimer (F1 = 2.448,P1 = 0.099;F2 = 0.024,P2 = 0.877). The effect of different doses of LPS and time of administration showed no influence on FDP (F1 = 0.106,P1 = 0.900;F2 = 0.013,P2 = 0.908), and no interactive effects were found (D- dimer:F = 0.002,P = 0.998; FDP:F = 0.582,P = 0.563).Conclusion LPS can induce TCP in mouse, but this effect may not related to the activation of coagulation system and excessive inflammatory response.

AIM: To establish a cell line of stable silencing of P2X7 receptor (P2X7R) expression through short hairpin RNA ( shRNA)-mediated interference in murine RAW264.7 macrophages, and to investigate the proliferation and apoptosis in the cell line.METHODS:Stable silencing of P2X7 R gene in the RAW264.7 cells was achieved by re-combinant shRNA plasmid targeting murine P2X7 R gene via liposome mediated transfection, followed by G418 selection. The efficacy of plasmid transfection and P2X7 R silencing in G418 resistant cells was verified by immunofluorescent micros-copy and real-time PCR, respectively.The proliferative activity was analyzed by CCK-8 assay and EdU cell proliferation as-say.The cell cycle distribution and apoptosis were evaluated by flow cytometry.RESULTS:The expression of P2X7 R at mRNA and protein levels was down-regulated by 80% in shP2X7 R group compared with negative control ( NC) plasmid transfection.In addition, P2X7 R-silencing cells exhibited higher proliferative activity compared with NC and wild-type RAW264.7 cells (P<0.05).Compared with NC cells, P2X7R silencing resulted in an increase in the phagocytosis of the cells ( P<0.05) .CONCLUSION:A cell line RAW264.7 of stable silencing of P2X7 R expression was successfully es-tablished.P2X7 R gene silencing stimulates the proliferation, and changes phagocytic function in murine RAW264.7 macro-phages.