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.

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.

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.

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.

OBJECTIVETo explore evaluation strategies for middle ear dysfunction in cleft palate patients, to optimize the diagnosis and treatment of this dysfunction, and ultimately to improve the comprehensive treatment of cleft palate.

METHODSThe relationship among abnormal tympanic types (B, C, and Anomaly), effusion rate, tympanic pressure, and hearing loss were analyzed. We collected relevant information on 469 ears of cleft palate patients and traced one-year longitudinal changes in the tympana of 124 ears from 62 patients with both cleft lip and cleft palate.

RESULTSThe effusion rates of cleft palate patients with type B, type C, and type Anomaly were 50.3% (97/193), 34.8% (8/23), and 20.9% (53/253), respectively. The tympanic pressure of the ears with and without effusion showed no significant difference (P>0.05). The hearing loss in type B cleft palate patients with middle ear effusion was worse than that in patients without effusion (P=0.001). However, the hearing loss in type Anomaly showed no difference (P>0.05). The constituent ratio of each tympanic type remained constant during the period between cheiloplasty and palatoplasty for cleft lip and palate patients (P>0.05).

CONCLUSIONCleft palate patients of all tympanic types may all suffer from middle ear effusion at different rates. Examination by centesis is suggested for ears with abnormal tympanic types. Early aggressive therapy is essential for type B cleft palate patients with middle ear effusion to avoid hearing loss. However, catheterization may be not necessary for type Anomaly patients, and conservative observation should be performed instead. Myringotomy with grommet insertion during palatoplasty does not delay treatment timing for patients with both cleft lip and cleft palateg.

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

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Mikulicz' Disease ; Salivary Gland Diseases

Autophagy is a common phenomenon which widely ex-ists in eukaryotic cells. Researches have shown that autophagy plays a critical role in maintaining cellular hemostasis, cell com-ponents update and keeping a normal physiological state. In re-cent years, the study has found that autophagy is closely related to the growth, the development of cardiovascular diseases and tumors. Further studies show that in different pathological condi-tions, autophagy could both promote angiogenesis and inhibit the formation of blood vessels. Therefore, it is particularly critical to elucidate the mechanisms of autophagy in the regulation of angio-genesis in different pathological conditions. The role of autophagy in cardiovascular diseases especially in the regulation of angio-genesis is discussed in this paper. Besides, the paper also in-cludes the discussion about the mechanisms of autophagy in the regulation angiogenesis, which may provide references for follow-up research and clinical treatment.

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.

Objective To detect the expressions of Lingo-1 and myelin associated protein in the white matter and hippocampus of ovariectomized rats after short-term estrogen replacement therapy in order to explore the possible mechanisms for the effects of estrogen on the brain myelin sheaths and cognitive function.Methods 24 middleaged (9-12 months) female Sprague-Dawley (SD) rats were bilaterally ovariectomized (OVX) and randomly divided into vehicle replacement (OVX+Veh) group and estrogen replacement (OVX+E) group.After one month ERT,The spatial learning and memory ability of all rats were assessed with Morris water maze.Then,10 rats were randomly selected from each group.The ultrastucture of myelin sheaths in the cerebral white matter and hippocampus were observed,and the protein expression of MBP and Lingo-1 were investigated with Western blot and immunohistochemical staining.Results The escape latencies of OVX+E rats in navigation test were significantly shorter than that of OVX+Veh rats(P＜0.05).The myelin sheaths in the white matter and hippocampus of OVX+Veh rats showed obviously degeneration.In the OVX+E group,the expression of MBP in the white matter and hippocampus was significantly higher than that of OVX+Veh group(P＜0.05),however,the expression of Lingo-1 was significantly lower than that of OVX +Veh group (P＜ 0.05).Conclusions One-month ERT has significant beneficial effects on the spatial learning capacity and myelin sheaths in the white matter and hippocampus.The protective effects may be related to estrogen-induced downregulation of the Lingo-1 expression in the white matter and hippocampus of rats.