OBJECTIVEHuman parainfluenza virus (HPIV) types 1, 2 and 3 are major viral pathogens responsible for upper and lower respiratory tract infections. In this study, a real-time RT-PCR was developed using multiplex primers-probe (HPIV-1, 2, 3) for the simultaneous detection of both HPIV1, HPIV2 and HPIV3 genomes.

METHODSOptimal primers and probes were designed using specialized software. The conditions for multiplex real-time RT-PCR had been optimized. The synthesis of RNA standards of HPIV1, 2, 3 were used a T7 RNA polymerase. Check the specificity sensitivities and stability of one step RT-PCR assay.

RESULTSObtained in a 10-fold dilution series assay demonstrate a high sensitivity of the assay with a lowest detection limit of 10 copies for HPIV1, 100 copies for HPIV2 and 100 copies for HPIV3.

CONCLUSIONThe assays demonstrates an improved sensitivity and scope of detecting HPIV1, 2, 3 viruses relative to routine antigen detection assays while the quantitative utility may facilitate investigation of the pre-diagnosis and respiratory virus pathogenesis.

Humans ; Oligonucleotides ; genetics ; Parainfluenza Virus 1, Human ; isolation & purification ; Parainfluenza Virus 2, Human ; isolation & purification ; Parainfluenza Virus 3, Human ; isolation & purification ; Real-Time Polymerase Chain Reaction ; methods

Objective To establish a model of acute gouty arthritis( AGA) in rats and observe its maintenance time. Methods The AGA model of rats was established by injecting monosodium urate ( MSU) at the concentration of 25 mg/mL into the ankle joint cavity. The rats were observed for 8 d at different time points. Skin temperature, degree of joint swelling, gait, inflammatory cells in synovial fluid, histopathological changes of synovial tissue and other indicators were observed to determine whether the modeling and maintenance time were successful. Results At 3 h after modeling, differ-ences in the swelling of ankle joint, increase of skin temperature, abnormal gait, the number of inflammatory cells in syno-vial fluid, synovial hyperplasia, capillary congestion, and disarrangement of synovial cells in the rats were observed in the saline group and the model group (P <0. 01). At 4 hours after modeling, the above mentioned inflammatory changes in the saline group were significantly reduced, compared with that at 3 h, showing a significant difference (P<0. 01), while the inflammatory changes of the model group were increased significantly compared with that at 3 hours ( P<0. 01 ) , and showed significant difference compared with the saline group (P<0. 01). At 24 h after modeling, the indexes in the rats of saline group returned to normal, but the inflammation of the model group was increased. At 48-72 h after modeling, the local inflammation such as ankle swelling, skin temperature, and abnormal gait of the rats in the model group reached a peak. The inflammation of the ankle joint in the model group was gradually reduced from 96 to 168 h after the model was established, but there were still significant differences in the indexes compared with the blank group (P<0. 01). At 192 h after modeling, the joint swelling, skin temperature and abnormal gait of the rats in the model group returned to normal, however, there were significant differences in the number of inflammatory cells and the pathological changes of synovial membrane compared with the blank group ( P<0. 01 ) . Conclusions A rat model of AGA can be successfully prepared and identified at 4 h after modeling by injection of MSU crystal suspension into the ankle joint cavity. This rat model of AGA can be maintained at least 168 hours after modeling.

OBJECTIVETo investigate the complex differences between high metastatic and low metastatic cells of the Adenoid cystic Carcinoma.

METHODSGene expression patterns were examined in high metastatic cell ACC-M strain and low metastatic ACC-2 strain with the method SSH (suppression subtractive hybridization).

RESULTSalthough extensive similarity was noted between the expression profiles, twelve genes were highly expressed of, in low metastatic cell ACC-2 tester, compared with driver, high metastatic cell ACC-M. These genes were cysteine-rich angiogenic-inducer protein (cyr61), chromosome7 clone RP11-52501, G protein, was family member Iferritin heavy polypeptide I, jumping translocation breakpoint, eukaryotic translation elongation, folate receptor, ribosomal proteins L7a, S21, P0 and other two novel genes-ACC metastasis-associated RNH and ACC metastasis-associated suspected protein. GenBank accession number were AF522024 and AF522025 respectively.

CONCLUSIONSthe result suggests that the obtainment of the ability of metastasis is related to the low expression or mutation of these genes. These data provide insight into the extent of expression differences underlying metastasis-related genes that may prove useful as diagnostic or prognostic markers.

Base Sequence ; Blotting, Northern ; Carcinoma, Adenoid Cystic ; genetics ; secondary ; Cell Line, Tumor ; Gene Expression Profiling ; Humans ; Molecular Sequence Data ; Reverse Transcriptase Polymerase Chain Reaction

Objective To analyze the peri-procedural complications of coronary rotational atherectomy. Methods A total of 107 consecutive patients with coronary artery disease received rotational atherectomy from January 2012 to December 2016 were enrolled retrospectively. The occurrence rate and the management of peri-procedural complications were summarized. Results ① The serum level of CK-MB and cTnI in patients who received rotational atherectomy were signifi cantly elevated compared with the level before the procedure,[4.1(2.4-14.1)U/L vs.1.5(1.0-2.1)U/L,P<0.001]and[0.28(0.11-1.11)ng/ml vs.0.01 (0.01-0.02) ng/ml, P<0.001], respectively. Of the 107 target lesions, 104 (97.2%) were classifi ed as type B2 or C lesions.②Peri-procedural complications of coronary rotational atherectomy occurred in 21 patients (19.6%). There were 5 cases (4.7%) of coronary slow fl ow, 1 case (0.9%) of severe coronary spasm, 9 cases (8.4%) of coronary dissection after rotational atherectomy and 1 case (0.9%) of side branch occlusion. Six patients (5.6%) had severe sinus bradycardia, but only 1 patient required temporary pacemaker. Burr entrapment happened in 2 patients (1.9%). Peripheral vascular complications occurred in 2 patients (1.9%) with 1 case of femoral artery hematoma at puncture site and the other 1 case of mediastinal hematoma. One patient (0.9%) developed contrast induced nephropathy after the procedure. ③All the peri-procedural complications of rotational atherectomy were benign. The procedure instant success rate was 98.1%, with no acute or sub-acute stent thrombosis, cardiac death, emergent coronary artery bypass graft occurred during hospitalization. Conclusions The complications associated with coronary rotational atherectomy are not rare, but severe complications are not common. The procedure could be safely performed with a high instant success rate.

BACKGROUNDHuman bocavirus (HBoV) is a parvovirus recently found to possibly cause respiratory tract disease in children and adults. This study investigated HBoV infection and its clinical characteristics in children younger than five years of age suffering from acute lower respiratory tract infection in Beijing Children's Hospital.

METHODSNasopharyngeal aspirates were collected from children suffering from acute lower respiratory tract infection during the winters of 2004 to 2006 (from November through the following February). HBoV was detected by polymerase chain reaction amplification and virus isolation and the amplification products were sequenced for identification.

RESULTSHBoV infection was detected in 16 of 333 study subjects. Coinfections with respiratory syncytial virus were detected in 3 of 16 HBoV positive patients with acute lower respiratory tract infection. The median age for HBoV positive children was 8 months (mean age, 17 months; range, 3 to 57 months). Among the HBoV positive children, 14 were younger than 3 years old, 9 were younger than 1 year old and 7 were younger than 6 months. These 16 positive HBoV children exhibited coughing and abnormal chest radiography findings and more than 60% of these children had wheezing and fever. Ten children were clinically diagnosed with pneumonia, 2 bronchiolitis, 2 acute bronchitis and 2 asthma. One child died.

CONCLUSIONSHBoV was detected in about 5% of children with acute lower respiratory infection seen in Beijing Children's Hospital. Further investigations regarding clinical and epidemiologic characteristics of HBoV infection are needed.

Bocavirus ; isolation & purification ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Parvoviridae Infections ; diagnosis ; etiology ; Polymerase Chain Reaction ; Respiratory Tract Infections ; diagnosis ; etiology

Objective To compare the setup accuracies of the surface-guided radiation therapy for nasopharyngeal carcinoma based on volume rendering technique(VRT)reference surface and digital imaging and communications in medicine(DICOM)reference surface.Methods Twenty patients with nasopharyngeal carcinoma who received radiotherapy using the customized AccuCushion with a thermoplastic fixation mask were selected.After the first cone beam CT(CBCT)-based image-guided radiotherapy of each patient was completed,the thermoplastic mask was removed to obtain the VRT reference surface based on optical surface monitoring system(OSMS),during which the patient position was maintained through instructions;the DICOM reference surface was got based on the body surface contour data extracted from the patient's localized CT images.After the completion of the 5th,10th,and 15th CBCT image-guided radiotherapy treatments,the patients'6 dimensions of setup errors were measured based on the 2 reference surfaces,respectively.Dimensional correlation analyses were carried out on the setup errors measured based on the 2 reference surfaces separately using Python's Stats tool to test the independence between the setup errors in each dimension.After independence test,Wilcoxon rank sum test was performed on the data of the setup errors based on each dimension measured with the 2 reference surfaces.Results The two sets of setup errors were independent of each other in all dimensions.The errors based on VRT reference surface were lower than those based on DICOM reference surface in 6 dimensions,while the difference between the 6-D errors of the two groups did not gradually become significant as the treatment progressed.Conclusion In the SGRT of nasopharyngeal carcinoma using a customized cushion combined with a thermoplastic mask,the setup accuracy and consistency based on VRT reference surface are both higher than those based on DICOM reference surface.

OBJECTIVETo analyze the postoperative complications in patients with endometrial carcinoma undergoing surgical operation in different modes and to explore the surgical safety of retroperitoneal lymph node dissection.

METHODSTwo hundred and nineteen patients with endometrial cancer treated in our hospital between May 2006 and April 2012 were included in this study. Their clinicopathological data were retrospectively analyzed. Among them, 65 patients received total abdominal hysterectomy and bilateral salpingo-oophorectomy (TAH+BSO group), 54 patients received TAH and BSO and pelvic lymph node dissection (PLX group), and 100 patients received TAH and BSO and PLX and para-aortic lymph node dissection (PALX group). The surgical procedures and postoperative complications in different operation modes were analyzed.

RESULTSThe operation time was (114.84 ± 6.45) min in the TAH+BSO group, (182.94 ± 6.62) min in the PLX group, and (188.27 ± 5.77) min in the PALX group. The operation time in the TAH+BSO group was significantly shorter than that in the PLX and PALX group (P < 0.001). The amount of blood loss was (222.97 ± 38.42) ml in the TAH+BSO group, (311.80 ± 21.62) ml in the PLX group, and (391.51 ± 53.20) ml in the PALX group. respectively. The amount of blood loss in the TAH+BSO was significantly less than that in the PLX and PALX group (P = 0.009). Lymphedema of the lower extremities was the most frequent complication of retroperitoneal lymph node dissection and the incidence rate was 31.8%. Lymphocyst was the second frequent complication, with an incidence rate of 27.3%. The incidence rate of ileus in the PALX group was significantly higher than that in the PLX group (P = 0.001). There were no significant differences in the incidence rate of lymphedema, lymphocyst and deep vein thrombosis between the PALX and PLX groups (P > 0.05).

CONCLUSIONSRetroperitoneal lymph node dissection is an acceptable operation mode, although slightly increasing the incidence of ileus, compared with the TAH+BSO group. It is needed to choose appropriate indication in order to decrease the post-operative complications.

Adult ; Blood Loss, Surgical ; Carcinoma, Endometrioid ; pathology ; surgery ; Endometrial Neoplasms ; pathology ; surgery ; Extremities ; Female ; Humans ; Hysterectomy ; adverse effects ; methods ; Lymph Node Excision ; adverse effects ; Lymph Nodes ; Lymphatic Metastasis ; Lymphedema ; etiology ; Lymphocele ; etiology ; Middle Aged ; Operative Time ; Ovariectomy ; adverse effects ; methods ; Pelvis ; Postoperative Complications ; epidemiology ; Retrospective Studies

The transcription factor X-box binding protein-1 (XBP1) plays a key role in unfolded protein reaction. This study was aimed to investigate the expression pattern and regulation of XBP1 in the mouse uterus during early pregnancy. The methods of immunohistochemistry (IHC) and real time quantitative RT-PCR were used to test XBP1 expression in early pregnancy, artificial decidualization, oestrous cycle and hormone-regulated mouse models. The results showed that XBP1 was spatiotemporally expressed in mouse uterus during early pregnancy. The XBP1 protein was mainly detected in the luminal and glandular epithelia on days 1-4 of pregnancy, and was strongly detected in the decidual area on days 5-8 of pregnancy. Similarly, XBP1 expression was also mainly expressed in decidual cells following artificial decidualization. During the oestrous cycle, Xbp1, Xbp1u, and Xbp1s mRNA was predominantly present in proestrus. In the ovariectomized uterus, the expression of XBP1 in luminal and glandular epithelia was up-regulated after estrogen treatment. These results suggest that XBP1 is associated with embryo implantation and decidualization during early pregnancy in mice, and the expression of XBP1 in luminal and glandular epithelia may be regulated by estrogen.
Animals ; Decidua ; Embryo Implantation ; Estrogens ; Female ; Mice ; Pregnancy ; RNA, Messenger/genetics* ; Uterus