The template-independent teminal transferase activity of Taq DNA polymerase results in an overhanging dA at the 3′end of its PCR products. The pGEMX vector constructed in this experiment forms a single overhanging dT at its 3′end as the result of cleavage with Xcm I restriction enzyme. This vector is very efficient for direct cloning of PCR product obtained by using Taq DNA polymerase.Recombinant colonies can be selected by Blue/white screening. Moreover,insertion fragment can be easily released from the vector simply with either BamH I or Hind III digestion.

OBJECTIVE To discuss the quality management of the recycled medical instrument.METHODS We executed the routine of recycled instrument seriously during our daily work,and gave strict measures during quality management such as callback,washing,classification,packaging,sterilization,handout,and inspection.RESULTS The recycled medical instrument had been used and managed for more than one year and the clinical work was ensured in progress smoothly and achieved satisfied results.CONCLUSIONS Actualizing the measure of quality control,perfecting the method of management and improving quality control could assure the safe and efficient use of recycled medical instrument and reach the standard of controlling hospital infection,and advancing medical nursing level.

Objective To investigate the testing capabilities of clinical transfusion laboratories in medical institutions in Beijing for the regulatory authorities to formulate administrative policies in this regard.Methods Experts assigned by Beijing Clinical Transfusion Quality Control Center made on-site inspections at the transfusion laboratories in medical institutions using quality control products.They recorded the complete testing process of the operators as well as the instruments,detection reagents in use and the testing results,with statistics and analysis made to the data so collected.Results The pass rate of these on-site inspections was lower than that of the external quality assessment.Some laboratories failed to complete the testing of the quality control products in time and the actual operations in some laboratories were inconsistent to the guidelines.55.9％ of level Ⅰ hospitals and 25.6％ of level Ⅱ hospitals were found with insufficient and inadequate instruments and process layout to meet the needs of clinical blood transfusion.Some of the technicians were found without sufficient trainings in their professional knowledge and basic skills,resulting in their poor competence against emergency cases and weakness in independent problem solving.In addition,the records of detection process and results were found to be substandard.Conclusions Transfusion laboratories in Beijing need to improve their testing capabilities in general.

OBJECTIVE To investigate the risk factors for nosocomial pneumonia(NP) caused by imipenem-resistant Acinetobacter baumannii(IRAB) and its antimicrobial susceptibility in vitro. METHODS The data of 34 cases of IRAB-NP were analyzed and 68 cases of NP caused by imipenem-susceptible A.baumannii(ISAB) were randomized as control.Antimicrobial susceptibility(MIC) was determined with the method of agar dilution. RESULTS The two independent factors associated with the development of IRAB-NP: previous fluoroquinolone(OR=5.738) and imipenem/meropenem(OR=7.129) use.The drug sensitivity test in vitro showed that these strains were multiresistant to commonly used antibiotics,and only ampicillin/sulbactam and cefoperazone/sulbactam whose resistance rate was less than 30%. CONCLUSIONS Previous imipenem/meropenem and fluoroquinolone use is independent risk factors for IRAB-NP.These strains are high drug resistant.

Objective To investigate the prevalence of serum Anti-sperm antibodies(AsAb) in group of prepubertal boys with cryptor-chidism before and after surgery,and explore whether it may be influenced by testicular location,orchiopexy,unilateral or bilateral cryptorchi-dism and deferens and(or) epididymis abnormality.Methods Serum AsAb was examined in 50 prepubertal boys with cryptorchidism and 50 boys with oblique inguinal hernia before and 6 months after surgery.Fifty boys who were normal were served as controls.IgG and IgM AsAb were determined by the immunodotting method.Results The preoperative and post-orchiopexy serum AsAb positive rates in cryptorchid boys were significantly higher than the preoperative and post-high ligation of hernial sac ones respectively in boys with oblique inguinal hernia(Pa0.05).Conclusion Cryptorchidism may induce an autoimmune response against sperm antigen in prepuberty independent of testicular location,orchiopexy,unilateral or bilateral cryptorchidism and deferens and(or) epididymis abnormality.

Abstract: Objective To analyze the changes of coagulation indicators in patients with hepatitis B complicated with Clonorchis sinensis (C. Sinensis), and provide reference value for diagnosis, drug using and prognosis monitoring. Methods　The patient samples were collected from the Third Affiliated Hospital of Sun Yat-sen University from June 2018 to February 2022 and divided into six groups. They were 40 healthy patients, 47 patients with simple chronic hepatitis B, 47 patients with post-hepatitis B liver cirrhosis, 40 patients with C. Sinensis mono-infection, 30 patients with chronic hepatitis B patients co-infected with C. Sinensis and 27 patients with post-hepatitis B liver cirrhosis patients coinfected with C. Sinensis. Four coagulation indexes, prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) and fibrinogen (FIB), were detected and compared among the groups. Results Compared with the healthy group, C. Sinensis mono-infection group had higher level of FIB and delayed PT, APTT; chronic hepatitis B group and chronic hepatitis B patients co-infected with C. Sinensis group had delayed PT, APTT, TT and significant lower FIB, these differences were statistically significant (all P<0.05). Compared with simple chronic hepatitis B group, post-hepatitis B liver cirrhosis and chronic hepatitis B patients co-infected with C. Sinensis had significant delayed PT, APTT, TT and lower FIB (all P<0.05). Compared with the post-hepatitis B liver cirrhosis and chronic hepatitis B patients co-infected with C. Sinensis, post-hepatitis B liver cirrhosis patients coinfected with C. Sinensis group had significant delayed PT, APTT, TT and lower FIB (all P<0.05). Conclusions The coinfection of C. Sinensis will further aggravate the coagulation dysfunction of HBV patients, leading to poor treatment and prognosis. HBV patients will have worse coagulation function in the process to post-hepatitis B liver cirrhosis; Therefore, it is important to pay attention to C. Sinensis co-infection when treating HBV patients, so that further guidance on clinical use and monitoring of prognosis can be provided.

Objective To evaluate the relationship between sternum protection and bone marrow suppression in postoperative radiotherapy for esophageal carcinoma. Methods Total of 98 postoperative patients with thoracic esophageal carcinoma were randomly divided into experimental group (52 cases) and control group (46 cases). All patients were given intensity-modulated radiotherapy (IMRT), with the dose of 50-50.4 Gy. The patients in experimental group were irradiated by 6 fields (4-fields in front, 2-fields behind) which were crossed to avoid direct exposure to the sternum. The patients in control group were irradiated by 5 fields (3-fields in front, 2-fields behind) with front-middle of the field passing through the sternum. Concurrently all patients received 2 cycles of cisplatin chemotherapy. Results Dmean, V20 and V30 of the sternum in the experimental group were (20.21 ±3.60) Gy, (40.78 ±7.19) % and (33.78 ±9.44) %, which were lower than those in the control group [(30.91±5.21) Gy, (81.01±4.81) %, (51.60±6.84) %], respectively (P<0.05). However, the volume and dose distribution of lung, spinal cord and heart were similar between the two groups (P> 0.05). Both the incidence rates of bone marrow suppression at 14th day and 35th day after radiotherapy were significantly higher in the control group (52.2%, 73.9%) than those in the experimental group (28.8 %, 50.0 %) (P< 0.05), and the incidence rate of bone marrow suppression at 7th day after radiotherapy was similar between the two groups. Conclusion Protecting and sketching for sternum in postoperative radiotherapy for esophageal carcinoma can reduce the incidence of bone marrow suppression effectively, which would not increase the radiation dose in the lung, heart and spinal cord.

Objective To study the expression of KAI1 and CD40 in gastric cancer and their correlation with clinicopathologic features and prognosis.Methods The expression of KAI1 and CD40 of 64 gastric cancer tissues was examinated by S-P immunohistochemieal methods.Results The overall positive rate of KAI1 was 22% in gastric cancer.Expression of KAI1 was positively related to the degree of tumor differentiation,it was negatively related to invasion depth and lymph node metastasis and the tumor clinical stage.The 5-year survival rate of cases with positive expression of KAII was significantly higher than that of cases with negative expression(X~2=42.426,P=0.000).The expression rate of CD40 in gastric cancer tissue was 34%.The expression of CD40 was significantly correlated with metastasis and clinical stage of gastric cancer.The 5-year survival rate of cases with positive expression of CD40 was significantly lower than that of cases with negative expression(X~2=4.841,P=0.028).There is no relationship between KAI1 and CD40 expression in gastric cancer.Conclusion KAI1 is associated with gastric cancer differentiation,invasion and metastasis.CD40 status helps to evaluate metastasis,and predict prognosis of gastric cancer.

OBJECTIVETo investigate the expression of long non-coding RNA-HOTAIR in prostate cancer cells and its effects on the growth and metastasis of the cells.

METHODSUsing quantitative reverse-transcription PCR (qRT-PCR), we determined the relative expression of HOTAIR in the normal human prostate epithelial cell line RWPE-I and prostate cancer cell lines PC-3 and DU145. We detected the effects of HOTAIR on the cell cycle and invasiveness of prostate cancer cells by RNA interference, flow cytometry, and Transwell mitration assay.

RESULTSThe expressions of HOTAIR in the PC3 and DU145 cells were increased 3.2 and 5.7 times, respectively, as compared with that in the normal RWPE-1 cells. After si-HOTAIR interference, the prostate cancer cells were arrested in the G2 phase and downregulated in the G1 phase. The invasive ability of the prostate cancer cells was evidently inhibited, with the inhibition rates of 32% and 44% of the PC3 cells and 43% and 34% of the DU145 cells for si-HOTAIR1 and si-HOTAIR2, respectively.

CONCLUSIONIncRNA HOTAIR is highly expressed in prostate cancer, which is associated with the growth and invasiveness of prostate cancer cells. HOTAIR is potentially a novel marker for the diagnosis and prognosis of prostate cancer.

Cell Cycle ; Cell Cycle Checkpoints ; Cell Division ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; G1 Phase ; G2 Phase ; Humans ; Male ; Neoplasm Invasiveness ; Prognosis ; Prostatic Neoplasms ; metabolism ; pathology ; RNA Interference ; RNA, Long Noncoding ; metabolism ; RNA, Untranslated ; metabolism

Objective To investigate the relationship between expression of active form of caspase-3 and cell cycle in Fas-mediated apoptosis of B lymphocytoma cell line MML-1. Methods MML-1 cells were incubated with agonistic anti-Fas antibody for different time,and cell apoptosis was induced.Cell apoptotic rates were analysed by flow cytometry,and sensitivity of MML-1 cells to apoptosis was determined.The expression of active form of caspase-3 was analysed by double staining with PI-Triton X and FITC-active caspase-3.Cyclin A,B_1 and E were selected as cell cycle markers for S,G_2/M and G_1 phase of MML-1 cells,and the expression of active form of caspase-3 was detected by flow cytometry. Results The cell apoptotic rate reached 56% after induction by Fas for 6 h.After induction by Fas for 4 h,the active form of caspase-3 was mainly expressed in cells of G_1 phase,while rarely in cells of S and G_2/M phase.Cells with negative cyclin A and B_1 and positive cyclin E expressed active form of caspase-3. Conclusion The expression of active form of caspase-3 in MML-1 cells mediated by Fas might be cell cycle dependent.Cells entering into late G_1 and early S phase first express active form of caspase-3,and their sensitivity to Fas-mediated apoptosis is the highest.