Objective To explore the inhibitory effect of astragalus polysaccharide (APS) on the proliferation of human erythroleukemia K562 cells and its mechanisms.Methods After K562 cells (purchased from Shanghai cell bank of chinese academy of science) were treated with different concentrations (0 mg/L,100 mg/L,200 mg/L and 400 mg/L) of APS.The influences of APS on the growth rate,doubling time and cell cycle distribution of K562 cells were observed by methyl thiazolyl tetra-zolium assay (MTF) and flow cytometry,respectively.Furthermore,the reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting assay were used to detect the expressions of Cyclin A,Cyclin B,Cyclin E and p21 gene at the mRNA and protein levels,respectively.Results MTT assay findings showed that,compared to the control group (0 mg/L APS),growth rates of K562 cells treated with 100 mg/L,200 mg/L and 400 mg/L APS decreased significantly (all P ＜ 0.01),and the doubling times lengthened significantly (all P ＜ 0.01).Flow cytometry findings revealed that,compared to the control group,the G1 phase cells in K562 cells of APS group increased significantly (P ＜0.01),while the S and G2/M phase cells decreased significantly (all P ＜ 0.01).RT-PCR and Western blotting results indicated that Cyclin B and Cyclin E expression of K562 cells at the mRNA and protein levels in the APS group were significantly lower than those of the control group(all P ＜ 0.01),whereas p21 expression was significantly enhanced at mRNA and protein levels (P ＜ 0.01),and Cyclin A expression was not significantly different at mRNA and protein levels between the 2 groups (all P ＞ 0.05).Conclusions APS could inhibit the proliferation of human erythroleukemia K562 cells.APS could inhibit the proliferation of K562 cells by down-regulating the expression of Cyclin B and Cyclin E and up-regulating the expression of p21.

BACKGROUND:Titanium aloy and stainless steel are two common internal fixation materials, but there are some difference in their therapeutic effects and biocompatibility. OBJECTIVE:To explore the therapeutic effects and biocompatibility of titanium aloy and stainless steel internal fixation materials for the treatment of spinal tuberculosis. METHODS:Seventy-one spinal tuberculosis patients, 35 males and 36 females, aged 17-81 were enroled. Among them, 35 patients received titanium aloy internal fixation, and the 36 patients underwent stainless steel internal fixation. At the end of 12-month folow-up, Cobb angle changes, therapeutic effect and Frankel grade were analyzed in the two groups. RESULTS AND CONCLUSION:Before treatment, there was no difference in the spinal kyphosis angle and Frankel grade between the two groups. At the last folow-up, the Frankel grade and Cobb angle were both improved in the two groups (P < 0.05), but there was stil no difference between the two groups. The cure rate was 97% (n=34) in the titanium aloy group and 92% in the stainless steel group (n=33), and no significant difference was found between the two groups. These two kinds of internal fixation materials exhibited good biocompatibility, and no infection and other adverse reactions occurred. These findings indicate that both titanium aloy and stainless steel as internal fixation materials have good biocompatibility and therapeutic effects.

Objective To explore the prognosis of elderly patients suffered from hospital-acquired pneumonia (HAP) by T cell subsets and clinical pulmonary infection score (CPIS).Methods A cohort of 125 elderly patients admitted in ICU & ED (Emergency Department) from Aug,2012 to Jul,2013 were enrolled for a prospective and observational study.The patients were divided into 3 groups:HAP survival group (n =50,group A),HAP death group (n =40,group B) and non-HAP group (n =35,control group).The criteria of exclusion were patients with auto-immune diseases,immunodeficiency,allergic disorders,malignancies,diabetes,trauma,surgical diseases,or patients with recent use of immunosuppressive agents or cyclooxygenase-inhibitors (Aspirin etc.).In the control group,patients with nosocomial pneumonia and other diseases afecting the CPIS were excluded.APACHE Ⅱ scores of all patients were recorded.Blood T cell subsets (including values of CD3,CD4 +,CD8 +,and CD4 +/CD8 +)were measured on the admission day,the 1st day of HAP onset and the 5th day after onset of HAP in HAP patients whereas these measurements were tested only on the admission day in controls.Meanwhile,the CPISs were recorded on the admission day,the 1st day of HAP onset and the 5th day after onset of HAP in HAP patients.Flow cytometer (FCM) was used to detect T cell subsets.Data of statistical analysis were represented as Mean ± SD.The significant differences in T cell subsets and CPIS between survival group and death group were analyzed by independent t test.The paired samples t test was employed in survival group and death group.Linear correlation analysis was made between CD4 +/CD8 + ratio and CPIS in survival and death groups,respectively.Results There were no significant differences in demographics and clinical features (including age,sex,length of stay,APACHE Ⅱ scores) of patients in survivors and non-survivors (P ＞ 0.05).The values of CDs (CD3,CD4 + and CD4 +/CD8 + ratio) between patients of control group and patients of HAP groups were not significantly different on the admission day (P ＞ 0.05).The values of CDs on the admission day were much lower than those on the 1 st day of HAP onset in both survivors and nonsurvivors (P ＜ 0.05).The values of CDs on the 5th day after onset of HAP were higher than those on the 1 st day of HAP onset in the survival group (P ＜ 0.05),while there were no significant differences in CDs between different intervals after HAP onset in the death group (P ＞ 0.05).There were no significant changes in values of CD8 + in any group (P ＞ 0.05).Both survivors and non-survivors had much higher CPIS values on the 1st day of HAP onset than those on the admission day (P ＜0.01).The survival group had higher CPIS on the 5th day after onset of HAP compared to the 1st day of HAP onset (P ＜0.01),while there was no significant change in the death group.Linear correlation analysis showed negative correlation between CD4 +/CD8 + ratio and CPIS on both the 1 st day of HAP onset (survival group:R =-0.740,P =0.004 ; death group:R =-0.613,P =0.035) and the 5th day after onset of HAP (survival group:R =-0.639,P =0.009; death group:R=-0.686,P=0.021).Conclusions The hospital-acquired pneumonia appears as an immune imbalance disorder.The difference in CDs is a promising objective tool,aiding in prediction of prognosis of HAP in the elderly,the lower the CDs,the higher severity.The CD4 + / CD8 + ratio showed a negative correlation with CPIS.Monitoring of T cell subsets and CPIS may provide clinical value for the treatment of hospital-acquired pneumonia in the elderly.

To study the value of lung ultrasound score (LUS) in assessing the clinical outcome of patients with ventilator-associated pneumonia (VAP).A total of 99 VAP patients were enrolled in a prospective study.All patients met the diagnostic criterion of VAP based on the 2013 guidelines and admitted into our ICU from Jun 2013 to Jun 2015.All parameters were recorded on the diagnostic day (day 1) and day 5,including LUS,clinical pulmonary infection score (CPIS),chest X ray (CXR),Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) score,Sequential Organ Failure Assessment (SOFA) score,etc.According to the CPIS,patients were divided into 2 groups(CPIS less than 6 and more or equal to 6).CPIS and LUS were similar on day 1 between two groups (P ＞ 0.05).However,on day 5,significant differences of CPIS and LUS were found between groups with CPIS ＜ 6 and CPIS≥6 (P =0.019 and P ＜ 0.001 respectively).LUS decreased on day 5 in CPIS ＜ 6 group and increased in CPIS ≥6 group.In CPIS ＜ 6 group,there was a positive correlation between LUS and CPIS on day 1 (r =0.375,P =0.003) and day 5 (r =0.590,P ＜ 0.001).CPIS ≥6 groupshowed the same trend on day 1 (r =0.484,P =0.002) and day 5 (r =0.407,P =0.011).LUS can be used to dynamically evaluate the clinical outcome of VAP.

BACKGROUND: Several studies have demonstrated that many American women who are at high risk of developing osteoporosis have higher levels of serum phosphorus. This indicates that some substances which can lower the serum level of phosphorus will supply a new and effective method to prevent and treat osteoporosis.OBJECTIVE: To observe the influences of porcine bone protein on bone mineral density (BMD) and serum levels of calcium and phosphorus in a rat model of osteoporosis.METHODS: Wistar rat models of osteoporosis were established by intramuscular injection of dexamethasone. Rat models were randomly divided into physiological saline, Jiegu Qili tablet, 50, 100, 200 mg/kg porcine bone protein groups. Rats that did not receive any treatments served as normal controls. After 12 weeks of treatment, serum was collected and serum levels of phosphorus and calcium were determined by biochemistry method. At the same time, tibia sections were made to determine tibial DMD by QDR-400 dual energy X-ray absorptiometry and to observe tibia marrow cavity by hematoxylin-eosin staining.RESULTS AND CONCLUSION: There was no significant difference in serum level of calcium among groups (P＞0.05).Compared with the physiological saline group, serum level of phosphorus in the 50, 100, 200 mg/kg porcine bone protein groups was significantly decreased (P ＜ 0.05). BMD was significantly higher in the 50, 100, 200 mg/kg porcine bone protein, Jiegu Qili tablet groups than in the physiological saline group (P ＜ 0.05). The tibia marrow cavity was smallest in the normal control group and largest in the physiological saline group. The tibia marrow cavity was larger in the 50, 100, 200 mg/kg porcine bone protein,Jiegu Qili tablet groups than in the physiological saline group. These results indicate that porcine bone protein cannot change the serum level of calcium, but it lowers serum level of phosphorus, and increases BMD, in a rat model of osteoporosis. However, the dose-dependent effect of porcine bone protein was not observed within the present experimental dosage. In addition, porcine bone protein can also reduce the marrow cavity of the tibia of rats with osteoporosis.

Objective To explore the clinical value of renal dynamic imaging and urinary N-acetyl-?-D-glucosaminidase(NAG),apoptosis DNA fragment(ADF) in evaluating the damage degree of hydronephrotic kidneys(HnK) in children with hydronephrosis.Methods Level of glomerular filtration rate(GFR) was detected in 41 children with congenital hydronephrosis by renal dynamic imaging,and urine NAG,ADF in pelvis in HnK and healthy kidneys (as controls) were detected by enzyme-linked immuno-sorbent assay(ELISA).Patholo-gic changes of HnK in 41 children were graded intoⅠ~Ⅴ according to Elder standard. And GFR,urinary NAG and ADF of HnK were divi-ded into subgroup according to pathologic changes ,at the same time statistical analysis was performed within each groups. And the correlations of pathologic grades with GFR,urinary NAG and ADF of HnK were analyzed.Results 1.Kindneys GFR in healthy kidneys and Hnk were (174.33?20.43)?10-3 L/min,(143.86?17.51)?10-3 L/min respectinely,and there was significant difference between healthy kidneys and Hnk (P0.05).3.There was significant negative correlation between GFR levels of HnK and pathologic grades(r=-0.814 P0.05).Conclusions For hydronephrotic kidneys,urinary NAG can eva-luate impaired nephric tubule whereas renal dynamic imaging may evaluate the damage level of glomeruli;urine ADF may not indicate the damage level of diseased kidneys in children with congenital hydronephrosis.

Objective To increase the awareness of virus-associated hemophagocytic syndrome.Methods Sixteen cases of virus-associated hemophagocytic syndrome were retrospectively analyzed.Results All presented with persistent high fever,cytopenia,hepato-splenomegaly,hepatic dysfunction,hypertriglyceridemia,hyperferritinemia,coagulopathy,hypofibrinogenemia,cytokine storm and a low natural killer cell activity.All patients had lymphohistiocytic accumulation in bone marrow.Treatment with high-dose gamma-globulin and high-dose methylprednisolone.Clinical symptoms and laboratory improved,and five patients died.Conclusion Aggressive early diagnosis and treatment are critical to improve survival.

Acute Disease ; Anemia ; etiology ; Child ; Fever ; etiology ; Humans ; Leukemia ; complications ; diagnosis ; therapy ; Male ; Patient Care ; Prognosis

Objective To study the protein expressions of Fas-associated death domain protein (FADD) and caspase-8 in rats with intracerebral hemorrhage ,and the effects of erythropoietin tp reveal the mechanism of neu-m-protection by EPO. Methods 126 male SD rats were randomly divided into three groups: Sham-operated group, intracerebral hemorrhage group, and EPO group. Each group was divided into seven subgroups according to the differ-ent time points (3,6,12,24,48,72 h and 7 d). The model of intracerebral hemorrage was established in rats by in-tracerebral injection of autogenous blood. The protein expressions of FADD and caspas-8 in rats tissue around the hemorrhagic and the normal brain tissue were detected by immunohistochemistry. Results The protein expressions of FADD and caspase-8 were increased [(4.66±0.46 ) and ( 15.89±1.81)] at 3 h after intracerebral hemorrhage, and peaked at 48 h [ (35.88±4.24 ) and (45.04±3.99)], the expressions of FADD and caspas-8 in the region around hematoma in EPO group significantly decreased compared with model group[ (3.92±0.64) and (28.24±1.90), (13.32±2.01 ) and (35.08±2.82)] at 3 h and 48 h. Conclusion The protein expressions of FADD and easpase-8 are markedly increased after intracerebral hemorrhage. EPO can protect the neurons by signifi-cantly reducing the expressions of FADD and caspase-8.

Objective To evaluate the significance of COLD-PCR in detecting KRAS mutation of pancreatic cancer and colorectal cancer patients. Methods First, set up COLD-PCR and compared the sensitivities of COLD-PCR/Sanger sequencing with PCR/Sanger sequencing using mixed cell lines ( KRAS wild-type cell line SW116 and KRAS mutant cell line SW480).Then, detected KRAS mutation of 20 formalin-fixed paraffin-embedded samples of pancreatic cancer and 39 formalin-fixed paraffin-embedded samples of colorectal cancer using PCR/Sanger sequencing and COLD-PCR/Sanger sequencing, respectively and compared the coincidence rate and consistency. Results The low detection limits of PCR/Sanger respectively. KRAS frequency detected by COLD-PCR/Sanger sequencing [75% (15/20)] in 20 cases of pancreatic cancer was higher than that detected by regular PCR/Sanger sequencing [40% ( 8/20 ) ,x2 =5.013, P < 0.05]. KRAS frequency detected by COLD-PCR/Sanger sequencing [44% (17/39)] in 39 cases of colorectal cancer was higher than that detected by regular PCR/Sanger sequencing [31% (12/39) ,x2 =1. 372, P = 0. 174]. The coincidence rate of these two methods was 0. 730 and the difference had no statistical significance. The coincidence rate of detecting KRAS mutation was 65% in pancreatic cancer and the results showed a good correlation between two methods and the two methods had bad agreement in diagnosis (Kappa = 0. 364, P < 0. 05 ). COLD-PCR/Sanger sequencing could detect more cases of KRAS mutations from pancreatic caner than regular PCR/Sanger sequencing. This was because there were many non-tumor cells in pancreatic tumor tissue and COLD-PCR/Sanger sequencing was more sensitive than regular PCR/Sanger sequencing. The coincidence rate of detecting KRAS mutations was 87% in colorectal cancer and the results were showed a good correlation between two methods and the two methods had substantical agreement in diagonsis ( Kappa = 0. 730, P < 0. 05 ) . Conclusion COLD-PCR/Sanger sequencing is highly sensitive to screen KRAS mutation in pancreatic cancer and colorectal cancer patients.