Objective To explore and evaluate the efficiency of the leucocyte depletion filter on eliminating tumor cells from blood, and its influence on the morphology and function of red blood cells. Methods Blood from operation field was collected, filtered, centrifuged and washed. Intestinal cancer cells (LOVO) and gastric cancer cells (SGC) were cultured in vitro and mixed with the collected blood. Blood contaminated with known number of tumor cells was then centrifuged and washed with cell retriever, and then the mixture was filtered with leucocyte depletion filter. The amount of residual tumor cells in filtered blood was counted after trypan blue staining before and after culturing. Unwashed and unfiltered blood containing tumor cells were used as control and were also cultured. The morphology and Na+-K+-ATPase activity of treated red blood cells was assessed. Results Tumor cells were detectable after only being washed with cell retriever and grew well after culturing for two weeks. No tumor cells were found after both washed with cell retriever and filtered with leucocyte depletion filter, and no tumor cell grew two weeks after culturing. No change was found in morphology and Na+-K+-ATPase activity of red blood cells after being washed and filtered. Conclusion Cell retriever is not able to eliminate tumor cells from blood completely. Further filtration with leucocyte depletion filter is capable of completely eliminating the residual tumor cells in blood. The morphology and Na+-K+-ATPase of red blood cells do not change after being washed with cell retriever and being filtered with leucocyte depletion filter.

Objective To investigate the effect of flavopiridol on the proliferation,invasiveness and apoptosis of human prostate cancer cell line LNCaP,and to explore the possibility of its application in clinical treatment.Methods MTT assay was used to detect cell proliferation,cell invasion in vitro was detected by Transwell assay,and flow cytometer was used to observe apoptosis.Results Flavopiridol inhibited the growth of LNCaP cells in a concentration-dependent and time-dependent way (P ＜ 0.05),and reduced the ability of invasion capacity.After treated by 10 nmol/L flavopiridol for 24 h,the apoptosis rate was increased significantly to (7.5±0.9) ％ compared with the control group [(5.3±0.5) ％] (P ＜ 0.05).Conclusion Flavopiridol can inhibit proliferation of LNCaP cells and induce apoptosis,which may be applicable for the treatment of prostate cancer.

Objective To investigate the expression of the enhancer of zeste homolog 2 (EZH2) gene and its significance in colorectal adenocarcinoma.Methods Immunohistochemistry and Western blot was used to assess the expression of EZH2 in human colorectal adenocarcinoma tissues,colorectal adenoma tissues and non-cancerous adjacent colorectal tissues.The relationships between EZH2 and each clinical pathology parameter were analyzed.Results The results of immunohistochemical trail showed that the expression rates of EZH2 in colorectal adenocarcinoma,colorectal adenoma and non-cancerous adjacent colorectal tissues were 80.56 ％ (87/108),62.5 ％ (25/40) and 5.00 ％ (2/40),respectively (P ＜ 0.05).Western blot revealed that the expression level of EZH2 in colorectal adenocarcinoma tissues,colorectal adenoma tissues and non-cancerous adjacent colorectal tissues level 0.549±0.145,0.283±0.023 and 0.107±0.022,respectively.The level in colorectal adenocarcinoma tissues (0.549±0.145) and colorectal adenoma (0.283±0.023) was significantly higher than that in non-cancerous adjacent colorectal tissues (0.107±0.022).Compared with colorectal adenoma tissues,level in colorectal adenocarcinoma tissues was significantly higher.There were significant differences among the three groups (F =20.113,P ＜ 0.05).The ratio of high expression level of EZH2 in colorectal adenocarcinoma tissues was closed related with tumorgenesis,differentiation,TNM staging and lymphatic metastsis (all P ＜ 0.05).However,no correlation was revealed between EZH2 expression and the age,gender (both P ＞ 0.05).Conclusion The expression of EZH2 may be associated with the tumorgenesis invasion,metastasis and progression of colorectal adenocarcinoma.

Objective To investigate the expression of protein interacting with Cα kinase 1 (PICK1) and its significance in colorectal adenocarcinoma.Methods The expression of PICK1 were detected by immunohistochemistry and Western blot methods in tissues of colorectal adenocarcinoma,colorectal adenoma,colorectal polyp and adjacent tissues.The correlation between PICK1 and clinical pathological data was explored.Results Immunohistochemical assay showed that the positive ratio of PICK1 protein was 72.5 ％ (74/102),and overexpressed in 31 cases (30.4 ％,31/102) with colorectal adenocarcinoma.The ratio of high expression level of PICK1 in colorectal adenoma tissues (22.2 ％,8/36) was significantly higher than that in the adjacent tissues (0,0/40) (P ＜ 0.05).The ratio of high expression level of PICK1 in colorectal polyp tissues (5.6 ％,2/36) was no statistically difference compared with that of the adjacent tissues (P ＞ 0.05).Western blot analysis revealed that the expression of PICK1 in colorectal adenocarcinoma (1.10±0.04) was significantly higher than that in the adjacent tissues (0.75±0.07) (P ＜ 0.05).The result showed significant correlation with the tumor location,the degree of differentiation,depth of invasion,TNM stages and lymph metastasis (all P ＜ 0.05).However,there is no correlation was revealed between PICK1 expression and the patients age,gender (both P ＞ 0.05).Conclusion The expression of PICK1 may be associated with the tumorgenesis,progression,invasion and metastasis of colorectal adenocarcinoma,which contributes to the prognosis of patients.

Objective To present the surgical technique of transumbilical laparoendoscopic singlesite pyeloplasty (LESS-P) for pediatric patients with ureteropelvic junction obstruction (UPJO).Methods Twenty-four pediatric patients with UPJO had transumbilical LESS-P performed by the same surgeon from June to December 2010.Sixteen patients were male and eight female aged from 2 to 62 months with an average age of 14 months.Eighteen patients had obstruction on the left side and six on the right side.Dismembered LESS-P was carried out with the Anderson-Hynes anastomosis where 5-0 sutures were uswed over a double J ureteric stent.Results All operations were successful.None was converted to open surgery and no additional sheath tube or incision besides the umbilicus was needed.No intraoperative complications occurred.Ectopic blood vessels were found in two patients during surgery.The mean operative time was 145 min,and the average blood loss about 10 ml.Abdominal drainage tubes remained for 2 -9 d after surgery.The mean postoperative hospital time was 7 d.Two patients had postoperative urinary fistula,which naturally disappeared at 4 and 7 d postoperation,respectively.Follow-up with ultrasound and diuretic renal scintigraphy found 23 patients had significantly decreased renal pelvis diameter,the remaining case showed no obvious change,but diuretic renography showed significantly improved excretion.ConclusionsPediatric transumbilical LESS-P could be safe and effective.LESS-P could achieve comparable clinic outcomes to standard laparoscopy.

Objective To investigate the expression of Dysadherin and analyze its role in renal clear cell carcinoma (RCCC).Methods RT-PCR and immunohistochemical were used to detect the expression of Dysadherin in 60 cases of fresh RCCC and 60 adjacent normal renal tissues(male 35,female 25; age 37-78,median age 61; ＞7 cm 24,≤7 cm 36; Ⅰ/Ⅱ 39,Ⅲ/Ⅳ 21).Results Dysadherin mRNA expression in RCCC tissues (2.0043±0.2890) was higher than that in adjacent normal renal tissues (0.8461 ±0.2479) (t =6.8020,P ＜ 0.05).Dysadherin expression was associated with nuclear grade.The expression of Dysadherin in nucleus grade Ⅲ and Ⅳ tumors were significantly higher than that in nucleus grade Ⅰ and Ⅱ tumors [the mRNA expression were 4.6224±0.3194,2.7780±0.2288,the positive rates of protein were 64.1％ (25/39),95.2 ％ (20/21) (t =6.5750,x2 =5.495,P ＜ 0.05)].There was no association between the expression of Dysadherin with sex (t =1.0530,x2 =0.023),age(t =0.0511,x2 =0.089) and tumor size (t =1.0330,x2 =0.370) (P ＞ 0.05).Conclusion In RCCC,Dysadherin expression is positively associated with tumor aggressiveness based on grading.It seems that Dysadherin may be a valuable prognostic marker in RCCC.

Objective To investigate the effects of sevoflurane pretreatment on the myocardial injury in patients undergoing cardiac valve replacement with cardiopulmonary bypass (CPB).Methods Twenty NYHA class Ⅱ or Ⅲ patients,aged ＜ 60 yr,undergoing cardiac valve replacement with CPB,were randomly divided into 2 groups (n =10 each):sevoflurane group (group S) and control group (group C).The patients were premeditated with intramuscular morphine and scopolamine.Anesthesia was induced with iv injection of midazolam 0.05-0.08 mg/kg,fentanyl 10-15 μg/kg and pipecuronium 0.08-0.10 mg/kg.Anesthesia was maintained with intermittent iv boluses of midazolam,fentanyl and pipecuronium and in addition sevoflurane was inhaled before aortic clamping and the end-tidal concentration was rapidly increased to 1.0％ and maintained at the level for 5 min in group S.Blood samples were taken from the central vein before skin incision (T1),immediately after aortic clamping (T2 ),at 0 and 30 min after aortic unclamping (T3-4),and at 2,6,12 and 24 h after operation (T5-8) for determination of the concentration of serum cardiac troponin Ⅰ (cTnI) and activities of creatine kinase (CK) and creatine kinase isoenzyme-MB (CK-MB).Myocardial specimens were taken from right auricle before aortic clamping and at 10 min after aortic unclamping for electron microscopic examination.Results The concentration of serum cTnI and activities of CK and CK-MB were significantly increased at T4-8 in both groups ( P ＜ 0.05).The serum cTnI concentration at T4-8 and the activities of CK and CK-MB at T8 were significantly lower in group S than in group C ( P ＜0.05).Different degrees of mitochondrial swelling were observed after aortic unclamping in both groups,but the changes were milder in group S than in group C.Conclusion Sevoflurane pretreatment can attenuate the myocardial injury in patients undergoing cardiac valve replacement with CPB.

Objective To construct the bait plasmid of HNP-3 mature peptide in yeast two-hybrid system and examine whether the recombinant bait plasmid has self-activating and toxicity effect.Methods Using RT-PCR technique,the cDNA fragments of HNP-3 mature peptide gene were amplified from the extracted RNA in cultured HL-60 cells.The fragment was firstly cloned into pBluescript-SK-II vector,confirmed by sequencing,then sub-cloned into the bait plasmid pGBKT7 and identified with PCR and sequence analysis techniques.The recombinant plasmid was introduced into the yeast cell AH109,and its self-activating and toxicity effect was tested by auxotrophic selective culture.Results DNA sequencing indicated that the inserted fragment in pBluescript-SK-II vector was HNP-3 mature peptide gene sequence,and the sub-cloned recombinant pGBKT7-HNP-3 was no mismatch.The recombinant bait plasmid didn't have self-activating effect and did not show toxicity to yeast AH109 cell.Conclusion The bait plasmid of HNP-3 mature peptide was constructed successfully.This was helpful for investigating the proteins interacting with HNP-3 mature peptide by yeast two-hybrid technique.

Objective:To investigate the targeted surveillance combined with cluster management on isolation protector use rate and hand hygiene qualification rate in caregivers of patients with respiratory tract multidrug-resistant bacterial infections.Methods:Second Hospital of Shanxi Medical University began to perform targeted surveillance combined with cluster management among caregivers of patients with multidrug-resistant bacterial infections in July 2019. The caregivers of 50 patients with respiratory tract multidrug-resistant bacterial infections who were admitted before targeted surveillance combined with cluster management were included in the pre-implementation group. The caregivers of 50 patients with respiratory tract multidrug-resistant bacterial infections who were admitted after targeted surveillance combined with cluster management were included in the post-implementation group. Multidrug-resistant bacterial infections in caregivers, isolation protector use rate, environmental disinfection and hand hygiene qualification rate were compared between before and after 6 months of targeted surveillance combined with cluster management. The application value of targeted surveillance combined with cluster management in the prevention of respiratory tract multidrug-resistant bacterial infections was analyzed.Results:The number of caregivers with respiratory tract multidrug-resistant bacterial infections in the post-implementation group was lower than that in the pre-implementation group (5 vs. 13, P < 0.05). The proportion of caregivers who wear a mask (94.11%), isolation clothes (80.39%) and gloves (98.03%) in the post-implementation group was significantly higher than that in the pre-implementation group (70.00%, 62.00%, 78.00%, χ2 = 10.027, 4.911, 9.683, all P < 0.05). The proportion of caregivers who were qualified in terms of hand hygiene (82.35%), object surface (76.47%), and keeping the air fresh in the post-implementation group was significantly higher than that in the pre-implementation group (64.00%, 58.00%, 64.00%, χ2 = 4.341, 3.915, 5.450, all P < 0.05). Conclusion:Targeted surveillance combined with cluster management can effectively help prevent respiratory tract multidrug-resistant bacterial infections in caregivers and increase isolation protector use rate and hand hygiene qualification rate.

Background::Alterations in the placental expression of glucose transporters (GLUTs), the crucial maternal-fetal nutrient transporters, have been found in women with hyperglycemia in pregnancy (HIP). However, there is still uncertainty about the underlying effect of the high-glucose environment on placental GLUTs expression in HIP.Methods::We quantitatively evaluated the activity of mammalian target of rapamycin (mTOR) and expression of GLUTs (GLUT1, GLUT3, and GLUT4) in the placenta of women with normal pregnancies (CTRL, n = 12) and pregnant women complicated with poorly controlled type 2 diabetes mellitus (T2DM, n = 12) by immunohistochemistry. In addition, BeWo cells were treated with different glucose concentrations to verify the regulation of hyperglycemia. Then, changes in the expression of GLUTs following the activation or suppression of the mTOR pathway were also assessed using MHY1485/rapamycin (RAPA) treatment or small interfering RNA (siRNA)-mediated silencing approaches. Moreover, we further explored the alteration and potential upstream regulatory role of methyltransferase-like 3 (METTL3) when exposed to hyperglycemia. Results::mTOR, phosphorylated mTOR (p-mTOR), and GLUT1 protein levels were upregulated in the placenta of women with T2DM compared with those CTRL. In BeWo cells, mTOR activity increased with increasing glucose concentration, and the expression of GLUT1, GLUT3, and GLUT4 as well as GLUT1 cell membrane translocation were upregulated by hyperglycemia to varying degrees. Both the drug-mediated and genetic depletion of mTOR signaling in BeWo cells suppressed GLUTs expression, whereas MHY1485-induced mTOR activation upregulated GLUTs expression. Additionally, high glucose levels upregulated METTL3 expression and nuclear translocation, and decreasing METTL3 levels suppressed GLUTs expression and mTOR activity and vice versa. Furthermore, in METTL3 knockdown BeWo cells, the inhibitory effect on GLUTs expression was eliminated by activating the mTOR signaling pathway using MHY1485. Conclusion::High-glucose environment-induced upregulation of METTL3 in trophoblasts regulates the expression of GLUTs through mTOR signaling, contributing to disordered nutrient transport in women with HIP.