OBJECTIVE: To construct the expressing vector of siRNA which can inhibit the Smad3 activity. METHODS: Sixty-four bases of 2 pair oligos for hairp in RNA expression which targeted Smad3 gene were chemically synthesized and annealed. pSUPER vector was linearized with BgL II and Hin d III treated with alkaline phosphatase (CIP). Anneled oligos were inserted into the downstream of the treated pSUPER's pol III H1 promoter to construct RNAi plasmid (pSUPER Smad3). Oligos with a scrambled sequence were used as a negative control. pSUPER Smad3 was transfected into human renal tubular epithelial cells (HKC). RESULTS: Recombinant pSUPER Smad3 vector was identified by the digestion with Eco R I and Hin d III, and confirmed by the sequencing analysis with T3 primer. Sixty-four bases had been inserted into the expected site. Furthermore, the insertion sequence was exactly corrected. The activity evaluation indicated that mRNA and protein of Smad3 but not Smad2 were inhibited by pSUPER Smad3 in HKC. CONCLUSION The pSUPER Smad3 system has been constructed successfully, and has high inhibition and specificity in vitro.
Epithelial Cells ; metabolism ; Humans ; Kidney Tubules ; cytology ; Plasmids ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Smad3 Protein ; antagonists & inhibitors ; genetics ; Transfection

Objective To assess the clinical effect of oblique lumbar interbody fusion (OLIF) combined with percutaneous pedicle screw fixation on computer navigation for lumbar spondylolisthesis.Methods Total 20 patients (8 males and 12 females with average age of 54.1± 12.3 years) with lumbar spondylolisthesis were enrolled in our study during Oct.2014 and May.2016.All patients were treated with OLIF combined with percutaneous pedicle screw fixation on computer navigation.Operation time,blood loss and complications were all recorded.Clinical and Radiographic evaluation were investigated on 1 week,3 months,6 months,12 months postoperatively and final follow-up.Visual analogue scale (VAS) for low back pain and leg pain,Oswestry disability index (ODI) for low back pain and the MOS item short form health survey (SF-36) were used to evaluate the clinical efficacy of surgery.Disc height,disc angle,lumbar lordosis and degree of upper vertebral slip of patients were investigated with X-ray.Cross-sectional area of intervertebral foramina was measured with three-dimensional CT and MRI.The cross-sectional area and sagittal diameter of the thecal sac were measured on T2-weighted axial and sagittal magnetic resonance images.Accuracy of pedicle screw placement was investigated with three-dimensional CT.Fusion rate was investigated with three-dimensional CT and Xray.Results All patients were followed for 12-30 months (22.9±4.8 months).The mean operation time was (119.0±23.8) min,the mean blood loss was (57.8±20.6) ml.VAS for low back pain,VAS for leg pain,and ODI were significantly improved from (6.7± 2.6),(6.3±2.7) and 50.5％±18.2％ preoperatively to (1.3±1.0),(0.8±1.0) and 14.0％±9.6％ at the latest follow-up.The SF-36 PCS and MCS scores were improved from (27.1 ± 13.9) and (51.0±22.7) preoperatively to (67.3± 18.9) and (81.2±14.1) at the latest follow-up.Disc height,disc angle,lumbar lordosis were significantly increased from (6.0±3.6) mm,1.8°±6.2° and 39.2°±8.4° preoperatively to (10.8± 1.7) mm,6.2°±3.5° and 45.0°±7.8° at the latest follow-up.Degree of upper vertebral slip of patients was reduced from 23.5％±7.4％ preoperatively to 4.2％±3.1％ at the latest follow-up.Cross-sectional area of intervertebral foramina in CT and MRI were significantly increased from (140.6±36.0) mm2 and (78.1±31.2) mm2 before surgery to (179.8±35.6) mm2 and (141.7±29.5) mm2 at 6 months after surgery.Cross-sectional area and sagittal diameter of thecal sac were significantly increased from (73.4±29.3) mm2 and (5.2±3.2) mm before surgery to (124.5±26.6) mm2 and (9.5±2.0) mm at 6 months after surgery.Accuracy of pedicle screw placement was 95％,and fusion rate was 100％ at 6 months after surgery.There were no severe vascular and nerve injuries.Conclusion OLIF combined with percutaneous pedicle screw fixation on computer navigation has good indirect decompression effect on lumbar spondylolisthesis,and was associated with high fusion rate.It can also effectively decrease the surgical trauma,improve the accuracy of pedicle screw placement,and increase disc height,disc angle and lumbar lordosis.

Objective To understand the current status and relative factors on knowledge, attitude, practice to contraception among college students in Beijing and to provide evidence for developing aimed contraceptive service and education in colleges. Methods Quantitative surveys were employed, questionnaire was conducted anonymously among 12 450 unmarried college students under informed consent. Results Among 12 129 students, acknowledgement of contraceptive knowledge among females was superior to males in college(P=0.000). Rates on the favor for unmarried sexual practice and 'sexual liberation' were 81.9% and 60.2% for males, comparing to 74.8% and 54.3% for females respectively. The prevalence rates for masturbation were 74.3% for males and 24.9% for females respectively. 18.4% of the male and 10.5% of the female students had ever experienced unmarried sexual intercourse. Among those students who had sexual experiences, 29.4% of the males had made their sex partners pregnant and 23.1% of the female students had experienced unexpected pregnancy. Statistical significance was found between males and females (P=0.000) in terms of attitudes and behaviors. 51.3% of the male and female students took contraceptive measures during the episode of first sexual intercourse. The excuses for not taking any measures would include: sexual intercourse happened incidentally, not interested in using condom, only one sexual experience would not cause pregnancy, etc. Conclusion The prevalence of unmarried sexual intercourse among college students had been rising. Among those who had experienced unmarried sexual intercourse, the rate of taking contraceptive measures was low and the incidence of unexpected pregnancy was high. It is urgent to strengthen the education and service programs on contraception among college students.

OBJECTIVETo compare the diagnostic value study the technique of ultrasonography, magnetic resonance cholangiopancreatography (MRCP) and endoscopic retrograde cholangiopancreatography (ERCP) in common duct stones.

METHODSThree hundreds and eighty-four patients suspected common duct stones from August 2005 to October 2007 were undergone abdominal ultrasonography, MRCP and ERCP.

RESULTSThere was stone in common duct in 370 and no stones in 14 of 384 patients. Ultrasonography indicated stones 268 cases, 8 false positive result was among them. MRCP diagnosed stones in 362 cases and false positive result in 6 cases, ERCP diagnosed stones 370 cases. The diagnostic accuracy of ultrasonography, MRCP and ERCP was 70.3% (260/370), 96.2% (356/370) and 100% respectively.

CONCLUSIONSThe diagnostic accuracy of ultrasonography for common duct stone was little higher, US should be used as a primary checking method. There was higher concordance between MRCP and ERCP for common duct stone. Combination of MRCP and ERCP can improve diagnostic accuracy of common duct stone.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Cholangiopancreatography, Endoscopic Retrograde ; Cholangiopancreatography, Magnetic Resonance ; Choledocholithiasis ; diagnosis ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Ultrasonography

As a result of the complex anatomy in upper cervical spine, the operative treatment of axis neoplasms is always complicated. Although the procedure for the second cervical vertebra (C2) surgery had been described previously in diverse approaches and reconstruction forms, each has its own limitations and restrictions that usually result in less satisfactory conclusions. The purpose of this study was to evaluate the operation efficacy for axis tumors by using a combined anterior (retropharyngeal) cervical and posterior approach in achieving total resection of C2 and circumferential reconstruction. Eight consecutive C2 tumor patients with mean age of 47.6 years in our institute sequentially underwent vertebra resection and fixation through aforementioned approach from Jan. 2006 to Dec. 2010. No surgical mortality or severe morbidity occurred in our group. In terms of complications, 2 cases developed transient difficulty in swallowing liquids (one of them experienced dysphonia) and 1 developed cerebrospinal fluid leakage (CSFL) that was resolved later. During a mean follow-up period of 31.9 months, the visual analogue scale (VAS) and Japanese orthopedic association (JOA) score revealed that the pain level and neurological function in all patients were improved postoperatively, and there was no evidence of fixation failure and local recurrence. It is concluded that the anterior cervical retropharyngeal approach permits a visible exposure to facilitate the C2 vertebra resection and perform an effective anterior reconstruction at the same time. The custom-made mesh cage applied in our cases can be acted as a firm and convenient implant in circumferential fixation.
Adult ; Axis, Cervical Vertebra ; diagnostic imaging ; surgery ; Combined Modality Therapy ; Female ; Humans ; Laminectomy ; methods ; Male ; Middle Aged ; Radiography ; Reconstructive Surgical Procedures ; methods ; Spinal Neoplasms ; diagnostic imaging ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo examine the expression of FLICE-inhibitory protein (FLIP) in juvenile idiopathic arthritis (JIA) and analyze its correlation with synovial inflammation.

METHODSThe expression of FLIP was assessed in 11 JIA and 3 normal synovial tissue samples by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. The cell types expressing FLIP were further characterized, and the correlation of FLIP expression with the degree of synovial inflammation, as well as the activity of caspase 8 was then analyzed.

RESULTSRT-PCR revealed the expression of FLIP mRNA in all 11 JIA samples, but not in 3 normal synovial tissues. In JIA, FLIP expression could be found in both the lining and sublining layers, mainly in the macrophage-like cells. Moreover, the expression of FLIP in JIA synovial tissues was positively correlated with the degree of synovial inflammation (r = 0.563, P < 0.05).

CONCLUSIONThe expression of antiapoptotic FLIP in JIA synovial tissue and its correlation to accumulation of inflammatory cells in synovial tissue suggests that FLIP potentially extends the lifespan of synovial cells and thus contributes to the progression of joint destruction.

Adolescent ; Arthritis, Juvenile ; metabolism ; pathology ; CASP8 and FADD-Like Apoptosis Regulating Protein ; genetics ; metabolism ; Caspase 8 ; metabolism ; Child ; Female ; Humans ; Inflammation ; metabolism ; pathology ; Male ; Protein Isoforms ; genetics ; metabolism ; Synovial Membrane ; cytology ; metabolism ; pathology

OBJECTIVELaparoscopic colorectal surgery is a skill-dependent procedure. The present study aims to analyze the learning curve of a properly trained surgeon, with basic laparoscopic techniques, to become skillful in performing laparoscopic colorectal operations.

METHODSA series of non-selective, consecutive 189 cases of laparoscopic colorectal surgery were accomplished, from December 2009 to February 2012, by one surgeon with years of skilled technique in laparoscopic cholecystectomy, rich experience in assisting laparoscopic colorectal surgery, and experience of approximately 180 procedures of gastric and colorectal surgery annually. 170 out of 189 procedures were radical operations for colorectal neoplasma, including right colectomies in 28 cases, left colectomies in 5 cases, sigmoidectomies in 28 cases, high Dixon procedures in 45 cases, low Dixon (total mesorectal excision, TME) procedures in 41 cases and Miles procedure in 23 cases. 19 other patients underwent combined procedures for multi-primary tumors or inflammatory enteritis. All these procedures were analyzed according to time span (the earlier half and later half) in respect to length of surgery, intraoperative blood loss, number of lymph nodes retrieved, intraoperative events and postoperative complications.

RESULTSFor radical right colectomy, the D2 dissection conducted in the earlier phase (n = 8) had the similar length of surgery, more blood loss and less LN retrieval, compared with the D3 dissection conducted in recent phase (n = 20). The earlier performed high Dixon procedures (n = 22) consumed longer time than the later procedures (n = 23) consumed, but with similar blood loss and LN retrieval. Low Dixon (TME) procedures showed significant differences in length of surgery and blood loss relative to time span. Recently performed simoidectomy and Miles procedures showed a trend of shorter time consumed compared with earlier performed procedures. Conversion ratio to open surgery was 1.05%. Adverse effects occurred in 8 cases of surgeries, including intestinal injury (3/189), insufficient distal margin (2/189), intraoperative bleeding (2/189) and vaginal injury (1/76). There was no operative death. Chief complications included urinary retention 5.82%, ileus 4.76%, anastomotic leak 4.24%, perineal infection 23.08% (6/26), wound dehiscence 2.65%, gastrointestinal bleeding 1.59%, peritoneal infection 1.06%. Surgery for distal rectum tended to have more complications, such as urinary retention, anastomotic leak and perineal infection. The later performed low Dixon procedures produced insignificantly fewer anastomotic leaks than those in the earlier phase.

CONCLUSIONSFor a trained surgeon with basic laparoscopic techniques, there are at least 15 - 25 cases of different procedures needed for him/her to become skilled to perform laparoscopic surgery. The learning curve should also depend on the annual number of colorectal surgeries.

Aged ; Colonic Diseases ; surgery ; Colorectal Neoplasms ; surgery ; Colorectal Surgery ; methods ; Female ; Humans ; Laparoscopy ; methods ; Learning Curve ; Male ; Middle Aged ; Postoperative Complications ; epidemiology ; Treatment Outcome

OBJECTIVETo investigate the safety and therapeutic effect of low dose (1000 U/m(2)) L-asparaginase (L-Asp) in the treatment of children with acute lymphoblastic leukemia (ALL).

METHODSSix patients were treated with low dose L-Asp after previously suffered severe side effects from standard dose L-Asp (5000 - 10,000 U/m(2)). Twenty-eight blood samples were obtained randomly from 5 of them. Plasma asparagine concentration was detected by reverse phase-high performance liquid chromatography (RP-HPLC).

RESULTSAll the patients treated with low dose L-Asp showed no any toxic symptoms. The plasma asparagine levels in the patients were all above 5 micromol/L except case 4 (4.91 micromol/L) before receiving L-Asp, and were all decreased below 0.5 micromol/L five days after receiving low dose L-Asp, except case 3 (3.70 micromol/L), the results being like that of receiving standard dose L-Asp.

CONCLUSIONLow dose L-Asp has definite efficacy for childhood ALL, while avoids serious side effects from standard dose L-Asp.

Adolescent ; Antineoplastic Agents ; administration & dosage ; adverse effects ; blood ; Asparaginase ; administration & dosage ; adverse effects ; blood ; Child ; Child, Preschool ; Female ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Treatment Outcome

OBJECTIVETo describe the current situation and trends of mean weight and height of the Chinese people.

METHODSSubjects of 71,971 households from 31 provinces were selected by stratified multi-stage cluster random sampling method. Height from 210 136 and weight from 202 749 subjects aged 2-79 years were used to describe their trends of development during 1992-2002. The height of children younger than 3 years of age was measured by infant height bed while the others were measured by metal height pole within 0.1 cm accuracy. Fasting body weight was measured to the nearest 0.1 kg.

RESULTSData on the mean height (cm) of Chinese children aged 2, 6, 11, 17 years were gathered which include: 1) in urban area: boys as 90.1 (2y), 118.4 (6y), 144.9 (11y), 170.2 (17y); girls as: 89.0 (2y), 117.0 (6y), 145.8 (11y), 158.6 (17y); 2) in rural area:boys as 87.6 (2y), 113.1 (6y), 139.2 (11y), 166.3 (17y) while girls as 86.2 (2y), 112.9 (6y), 140.0 (11y), 157.0 (17y). When compared to the results from 1992, the average height increased among all age groups. The values of increase among different groups aged 2-5 years were seen as: 3.6 cm of boys in urban and 3.0 cm of boys in rural areas while 3.8 cm of girls in urban and 3.0 cm of girls in rural areas. Among the aged 6-19 year groups, the average increasing values of height were 3.1 cm, 3.4 cm, 2.8 cm and 3.1 cm of city boys, rural boys, city girls and rural girls, respectively versus adults as 0.7 cm, 0.9 cm, 0.6 cm and 1.0 cm, respectively. The mean weight (kg) of Chinese children aged 2, 6, 11, 17 years were listed as below: boys in urban area: 13.5 (2y), 22.2 (6y), 37.4 (11y), 58.7 (17y); girls in urban area: 12.7 (2y), 21.2 (6y), 36.7 (11y), 51.9 (17y); boys in rural area: 12.8 (2y), 19.4 (6y), 31.9 (11y), 54.9 (17y); girls in rural area: 11.9 (2y), 18.7 (6y), 31.8 (11y), 51.2 (17y). The average weights of all age groups on 2002 were all higher than their same age counterparts in 1992. The average increasing values of weight among children aged 2-5 years were 1.0 kg, 0.6 kg, 0.8 kg and 0.5 kg for city boys, rural boys, city girls and rural girls, respectively. Among the aged 6-19 year groups, they were 2.0 kg, 1.5 kg, 1.4 kg and 1.1 kg, while among the adults aged 20-70 years, they were 3.2 kg, 3.0 kg, 1.6 kg and 2.9 kg, respectively.

CONCLUSIONBoth mean weight and height had a remarkable increase and the physical status improved among all age groups for the last ten years.

Adolescent ; Adult ; Age Distribution ; Aged ; Asian Continental Ancestry Group ; statistics & numerical data ; Body Height ; Body Weight ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Middle Aged ; Rural Population ; statistics & numerical data ; Sex Distribution ; Urban Population ; statistics & numerical data ; Young Adult

The aim of study was to investigate the effect of acute lymphoblastic leukemia (ALL) children bone marrow mesenchymal stem cells (MSC) on resistance of K562/A02 cells and its mechanism. MSC obtained from bone marrow of AL children were cultured and identified. The co-culture of MSC and K562/A02 and the culture of K562/A02 cell suspension alone was performed, of which 2 kinds of cells were treated with same concentration of adriamycin (ADM), and the rate of apoptosis was detected by flow cytometry, bcl-2 and bax of K562/A02 were detected by RT-PCR, while mdr1 gene level was detected by FQ-PCR. The results indicated that the MSC separation and proliferation were viable and steady. The apoptosis rate of the K562/A02 cells co-cultured with MSC was 1.97 ± 0.11%, while apoptosis rate of the K562/A02 cells cultured alone was 8.38 ± 0.29%, there was significant difference (p < 0.05). As compared with the K562/A02 cells cultured alone, the bcl-2 gene expression in K562/A02 cells co-cultured with MSC obviously increased; ratio of bcl-2/bax was obviously enhanced. The mdr1 gene level in K562/A02 co-cultured with MSC was no statistical different from K562/A02 cultured alone (p > 0.05), which suggested that adhesion co-cultured with MSC did not induce mdr1 expression higher than the culture of suspension. It is concluded that the MSC of ALL children can escape the leukemia cells from proapoptotic effect of drugs, the resistance of K562/A02 to ADM may be involved in enhancement of bcl-2 gene expression of K562/A02 cells co-cultured with MSC, but not in relation to mdr1 gene in K562/A02 cells themselves.
ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Bone Marrow Cells ; drug effects ; Child ; Child, Preschool ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Female ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Male ; Mesenchymal Stromal Cells ; drug effects ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; bcl-2-Associated X Protein ; genetics