Polymerase Chain Reaction (PCR)in recent years is a new disease detection methods basedon molecular biology level.It has the advantage of sensitive,specific,trace,simple,fast,but in the operation due to multiple factors,quality control is not easy to control.Especially in the first stage of quality control,it engages alot of operations and is weakest stage such needs most concern.It is necessary and important to have a good quality control;it not only improve the reliability and accu-racy of PCR testing,treatment of molecular biology,the professional level of work staff,but also reduce patient disputes and increase technology access,acceptance of the PCR laboratory.In this paper,the author from the factors which influence the first stage of PCR:whether correctly fill PCR test application form,patient preparation,handling,transportation,storage of good sample,unqualified specimen rejection,the requirement of laboratory technicians,calibration and maintenance on the in-struments;quality check on reagents,consumables quality inspection,standard operating procedures (SOP)writing,and la-boratory safety.Hope this paper can do a good reference for other same profession.

Protein-protein interactions (PPI) are involved in a variety of biological processes, including cell-to-cell interactions, metabolism and development control. The misregulation, post-translational modification and interference of PPI are related to a variety of human diseases, making the regulation of these interactions a very attractive field of drug discovery. In recent years, the interaction between MDM2 and p53 has become a research hotspot, which plays an important role in the treatment of tumors. But unfortunately there are no such inhibitors approved all over the world. In this view, recent advances of MDM2-p53 inhibitors were briefly described and its inhibitors with potential therapeutic activities in clinical studies were introduced.

OBJECTIVETo investigate the administration of far-lateral approach in lesions located anterior or anterolateral to brain stem and upper cervical spinal cord.

METHODSTwenty-three patients underwent far lateral approach, including 12 retrocondylar approach, 5 partial transcondylar approach, 3 transfacetal and partial transcondylar approach, 2 transtubercular approach and 1 complete transcondylar approach.

RESULTSTotal tumor removal was achieved in 15 patients, subtotal removal in 5 patients, 3 vertibral artery aneurysms were clipped successfully, 3 patients were given occipitalcervical fusion. There was no operative mortality. The most frequent complications were lower cranial nerve deficit, CSF leakage, injury to vertibral artery, and ischemia of brain stem, cerebellum or spinal cord. No patient presented clinical instability of the occipitocervical junction after surgery.

CONCLUSIONSThe far-lateral approach is an ideal approach to structures located ventral to cranial-cervicle junction. But some of the surgical steps are technically difficult and carry some degree of risk. The choice of approach depends on the pathological feature and degree of exposure required for effective surgical treatment. Bone removal should be quantified for individual lesion. The approach may be limited to less aggressive steps, while still achieving significant exposure and surgical space.

Adolescent ; Adult ; Aneurysm ; surgery ; Basilar Artery ; surgery ; Brain Neoplasms ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Microsurgery ; Middle Aged ; Neurosurgical Procedures ; methods ; Treatment Outcome ; Vertebral Artery ; surgery

OBJECTIVETo demonstrate the chemical constituents of Alyxia sinensis.

METHODThe constituents were isolated by column chromatography and identified by advanced physical and spectral analysis.

RESULTEight compounds have been isolated and elucidated as bauereny acetate(18), scopletin(19), liriodendrin(20), pinoresinol-di-O-beta-D-glucopyranoside(21), daucosterol(22), flaxetin(23), esculin(24), aseculin(25).

CONCLUSIONThese compounds were found from the plant for the first time, and compound 20,21,23-25 were found from Alyxiae genis for the first time, and compound 18 is firstly been isolated from natural source.

Apocynaceae ; chemistry ; Esculin ; chemistry ; isolation & purification ; Furans ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Triterpenes ; chemistry ; isolation & purification

Objective To study the mlationship between dietary soy isoflavones and blood lipids among residents of 40-65 years old,in Guangzhou.Methods Dietary soy isoflavones and other nutrients intakes were assessed with quantitative food frequency questionnaire(FFQ).Total cholesterol(TC),triglycerides(TG),HDL cholesterol(HDL-C)and LDL cholesterol(LDL-C)in plasma were measured with colorimetry.Results Ranges of dietary soy isoflavones intake among 134 males and 261 females were fxom 0 mg/day to 61.96 mg/day and 0 mg/day to 82.52 mg/day,with means of 11.95 mg/day,14.90 mg/day,respectively.After adjusted for total energy intake and fat percent energy,difiefences of TC,LDL-C in total population and TC in women were statistically significant between groups(P value was 0.002,0.008,0.004,respectively) and dose-effect relationships(P value was <0.001.0.012.0.001,respectively)were observed between dietary soy isoflavones intake and the upper mentioned three indices.Compared with the low-intake group,tbese three indices lowered 7.06%,10.13%and 7.48%,respectively in high-intake group.Critical significance of LDL-C was observed both in women and men between groups.Further controlled for age,BMI and WHR,no obvious change of the results was observed.Conclusion Moderate intakes of soy isoflavone as part of a regular diet seemed to be associated with favorable blood lipid levels.

OBJECTIVETo investigate the clinical application value of 8.5/11.5 F transurethral seminal vesiculoscopy in the diagnosis and treatment of refractory hematospermia.

METHODSWe retrospectively analyzed 78 cases of refractory hematospermia diagnosed and treated by 8.5/11.5 F transurethral seminal vesiculoscopy from June 2012 to June 2014. The patients underwent serum PSA examination, transrectal ultrasonography, seminal vesicle ultrasonography, and pelvis CT or MRI before surgery, and all received transurethral seminal vesiculoscopy under the 8.5/11.5 F rigid ureteroscope.

RESULTSOperations were all successfully accomplished, which revealed abnormal opening of the ejaculatory duct in 5 cases, mucosal inflammatory hyperemia in the prostatic utricle and seminal vesicle in 78, dark red mucilage substance in the seminal vesicle in 34, seminal vesicle stones in 19, small polyp in the seminal vesicle in 2, and ejaculatory duct or seminal vesicle cyst in 4. All the patients received symptomatic treatment during the surgery. After surgery, hematouria was found in 13 cases, which disappeared within 2 weeks, pelvic hematoma in 1 case, which was cured by conservative treatment within 3 months, and epididymitis in 2 cases, which was controlled by anti-infection treatment. Hematospermia recurred in 3 cases during the 1-year postoperative follow-up.

CONCLUSION8.5/11.5 F transurethral seminal vesiculoscopy, with its advantages of easy operation, wide field of vision, large channel for operation, and few complications, deserves general clinical application in the diagnosis and treatment of refractory hematospermia.

Calculi ; Ejaculatory Ducts ; Endoscopy ; methods ; Epididymitis ; etiology ; Hemospermia ; diagnosis ; therapy ; Humans ; Magnetic Resonance Imaging ; Male ; Postoperative Period ; Recurrence ; Retrospective Studies ; Seminal Vesicles ; Tomography, X-Ray Computed ; Urethra

Objective To analyze the CT and MRI characteristics of the pineal region tumors (PRTs) to improve the accuracy of diagnosis and differential diagnosis.Methods The MRI and CT findings of 28 patients with PRTs proved by operation and pathology were retrospectively analyzed.Eight of them were performed preoperative 1H-MRS and DWI examination and thier data were analyzed too.Results of these 28 patients,11 were diagnosed as having germinoma,4 teratoma,3 astrocytoma,3 meningioma,2 pinealocytoma and 1 epidermoid cyst,1 pineoblastoma,1 hamartoma and 1 dermoidcyst.The main imaging characteristics were cystic tumors in the pineal region,variational density/signal intensity with few tumors having their specified morphology,density/signal intensity and enhancement style.The germinomas and the pineoblastomas were mainly homogeneous solid masses with symmetrical density/signal.The astrocytoma was cystic-solid,mostly,with changing shapes and odds density/signal.The mature teratoma showed blending density/signal.The 1H-MRS showed typical high Cho peak and low NAA peak in the astrocytoma; no NAA peak was found in the meningeoma; high Cho peak and no Lip peak were found in the pineoblastoma; the germinoma was noted having Lip peak.Conclusion Observation on the changes of the morphologic features,density/signal intensity and enhancement style can help in the differential diagnosis of PRTs.DWI and 1H-MRS examinations can improve the accuracy of differential diagnosis to some extent,but some PRTs remain to be non-differentiable.

Primitive trigeminal artery (PTA) and primitive otic artery (POA) is a very rare entity in adult life. We present a case of PTA and POA associated with a giant unruptured cavernous aneurysm in a 54-year-old woman. The PTA and the POA arose from the sac of the aneurysm directly, which greatly complicated endovascular therapy management.
*Cerebral Angiography ; Cerebral Arteries/*abnormalities ; Diagnosis, Differential ; Embolization, Therapeutic ; Female ; Humans ; Intracranial Aneurysm/*radiography/therapy ; Middle Aged

OBJECTIVETo observe the change of activation and proliferation ability of rat T-lymphocytes after suppress ICOS gene expression by RNA interference.

METHODSFour interference sites targeting at rat ICOS gene were designed and four pairs of oligonucleotide fragments were cloned into the pSilencer 4.1-CMV neo plasmid vectors then transfected into rat lymphocytes with cationic liposome. The expression of mRNA and protein of ICOS was detected by RT-PCR and flow cytometry. The alteration of lymphocyte proliferation ability was evaluated by mix lymphocyte reaction, and the secretion levels of IFN-gamma and IL-4 were measured by ELISA procedure.

RESULTSAfter transfection, the expression of mRNA and protein of ICOS in test groups were lower than that in control groups (P < 0.05). The ability of T-lymphocytes in proliferation was poor and the levels of IFN-gamma and IL-4 were reduced with ICOS gene shut down.

CONCLUSIONSRNA interference plasmid vector can suppress ICOS expression in rat T-lymphocytes significantly, and it may be useful for further study on transplantation immunity tolerance.

Animals ; Antigens, Differentiation, T-Lymphocyte ; genetics ; metabolism ; Cell Proliferation ; Cells, Cultured ; Female ; Genetic Vectors ; Inducible T-Cell Co-Stimulator Protein ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Lymphocyte Activation ; Male ; RNA Interference ; Rats ; Rats, Sprague-Dawley ; T-Lymphocytes ; immunology ; metabolism ; Transfection

OBJECTIVETo detect the expression of CBX7 in human glioma and investigate the potential regulatory effect of abnormally expressed microRNAs on CBX7 expression.

METHODSSemi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot were applied to detect the expression pattern of CBX7 in 2 human normal brain tissues, 9 glioma tissues, and 3 glioma cell lines. Miranda algorithm and Ensemble Machine Learning algorithm were combined to predict miRNAs that target human CBX7. The expression of miR-9 in those tissues and cell lines were detected by real-time PCR. After miR-9 overexpression in 293ET and miR-9 knock-down in T98G, luciferase assay and Western blot were used to confirm the effect of miR-9 on CBX7 expression. MTT assay and flow cytometry were applied to detect the effect of miR-9 knock-down on T98G cells.

RESULTSNo obvious difference in the CBX7 mRNA level between normal and tumor tissues was observed, while the protein level of CBX7 was abrogated or markedly reduced in glioma tissues and cell lines. Several miRNAs including miR-9 may target CBX7 by bioinformatics prediction. MiR-9 was up-regulated in glioma tissues and cell lines. In 293ET cell, luciferase activity of CBX7-3'UTR reporter was decreased to 24% after miR-9 overexpression. After miR-9 knock-down in T98G cell, the luciferase activity was increased by 1.8 fold and there was no change of CBX7 mRNA, while the protein level of endogenous CBX7 was significantly increased. The number of survival T98G cells increased and cells in G1 phase decreased after miR-9 knock-down.

CONCLUSIONIn human glioma, CBX7 is down-regulated by the inhibition of miR-9 at posttranscriptional level.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Algorithms ; Blotting, Western ; Brain ; metabolism ; Cell Line ; Cell Line, Tumor ; Cell Proliferation ; Child ; Female ; Flow Cytometry ; Glioma ; genetics ; metabolism ; Humans ; In Vitro Techniques ; Male ; MicroRNAs ; genetics ; physiology ; Middle Aged ; Polycomb Repressive Complex 1 ; Repressor Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult