Objective: Detecting the expression and mutation of human telomeric repeat binding factor (hTRF1) in 10 malignant hematopoietic cell line cells on the base of determining its genomic structure and its four pseudogenes to clarify if hTRF1 mutation is one of the factors of the activation of telomerase. Methods: hTRF1cDNA sequences were obtained from GenBank, its genome structure and pseudogenes were forecasted by BLAST and other biology information programs and then testified by sequencing. Real-time RT-PCR was used to detect the expression of hTRF1mRNA in 10 cell line cells, including myelogenous leukemia cell lines K562, HL-60, U-937, NB4, THP-1, HEL and Dami; lymphoblastic leukemia cell lines 6T-CEM, Jurkat and Raji. Telomerase activities of cells were detected by using telomeric repeat amplification (TRAP)-ELISA protocol. PCR and sequencing were used to detect mutation of each exon of hTRF1 in 10 cell line cells. Results: hTRF1 gene, mapped to 8q13, was divided into 10 exons and spans 38.6 kb. Four processed pseudogenes of hTRF1 located on chromosome 13, 18, 21 and X respectively, was named as ΨhTRF1-13, ΨhTRF1-18, ΨhTRF1-21 and ΨhTRF1-X respectively. All cell line cells showed positive telomerase activity. The expression of hTRF1 was significantly lower in malignant hematopoietic cell lines cells (0.0338, 0.0108~0.0749) than in normal mononuclear cells (0.0493, 0.0369~0.128) (P=0.004). But no significant mutation was found in all exons of hTRF1 in 10 cell line cells. Four variants were found in part of intron 1, 2 and 8 of hTRF1. Their infection on gene function is unknown and needs further studies. Conclusion: hTRF1 mutation is probably not one of the main factors for telomerase activation in malignant hematopoietic disease.

Objective: To evaluate the safety and clinical efficacy of segmental radiofrequency ablation of pulmonary vein (PV) ostia for patients with refractory paroxysmal atrial fibrillation (AF) under multi-slice spiral computed tomography (MSCT) guidance before the procedure. Methods: A series of 58 consecutive patients with refractory paroxysmal AF were enrolled to undergo segmental radiofrequency ablation of PV ostia. The 36 male and 22 female patients with mean age of (57.4±9.5) (32~79) years and no obvious organic heart disease. Before ablation, patients received MSCT to generate 3-dimentional image of the left atrium (LA) and proximal PVs. Patients then underwent segmental radiofrequency ablation of PV ostia using PV circular mapping catheter manipulated several times to ensure complete isolation between PVs and LA. Results: No complications occurred during the procedure. One patient developed delayed cardiac tamponade, which was drained percutaneously. The mean follow-up time was (17.1±9.3) months. Forty-one patients (95%) experienced improved quality of life one month after the procedure. Thirty-six patients (83%) showed stable sinus rhythm, while 10 patients (23%) required additional anti-arrhythmic drugs. AF returned≥1 time in 6 (14%) patients who underwent anti-arrhythmic drug therapy, but the number of episodes was less than that before the procedure. However, one patient experienced recurrent episodes of atrial flutter. Conclusion: It is safe and effective to perform segmental radiofrequency ablation of PV ostia for patients with refractory paroxysmal AF using MSCT guidance mappening.

Objective: To compare a new device (Innocor) for non-invasive measurement of cardiac output (CO) by foreign gas rebreathing method with conventional techniques used in the measurements of cardiac function. Methods: Cardiac outputs measured by Innocor (CO_RB) were compared with CO obtained by echocardiography (CO_EC), Swan-Ganz thermodilution (CO_TD), and left ventricle radiography (CO_LVR) in 34 patients subjected to cardiac catheterization. Values obtained from the four methods were analyzed by linear regression and paired values were compared by the method of Bland and Altman in SPSS. Results: There was strong positive correlation (r=0.94) between Innocor cardiac output values and the corresponding values obtained by thermodilution and between CO_EC and CO_LVR values. Thermodilution appears to overestimate cardiac output when compared to the values obtained with Innocor by (0.66±0.22) L/min (P<0.0001). There was no correlation between data obtained by Innocor and the corresponding CO_EC and CO_LVR values. Conclusion: Innocor CO_RB is an easy, safe and well established method for non-invasive measurement of cardiac output with good prospects for clinical application in heart disease patients.

Objective: This study was designed to detect the expression of bcl-2 and p53 proteins in colorectal carcinomas and to determine their association with the patient survival and stage of the diseases. Methods: Immunohistochemistry method was used to detect the expression of bcl-2 and p53 proteins in 93 cases of colorectal carcinoma. The stain results were obtained by analyzing the clinic-pathological characteristics of patients. Results: Fifty-seven percent (53/93) of the colorectal carcinomas were bcl-2 protein positive. The positive rate of bcl-2 protein in lymph node involvement cases was lower (15/37) than the cases without node involvement (38/58, P<0.01). The positive rate of p53 protein was 43% (40/93) in colon-rectum carcinomas. No significant correlation was observed between p53 protein expression and clinic-pathological manifestations (P>0.05) but the survival was significantly worse (P=0.0001) in the p53 protein positive cases. Neither bcl-2 nor p53 alone was correlated with stage of the disease. When combined bcl-2/p53 status was analyzed, a group with bcl-2(+) and p53(−) had the best prognosis. This group was significantly associated with earlier Dukes’ stages (P=0.1763). In multivariate Cox regression analysis, lymph node involvement and p53 protein expression were two independent factors correlated with survival time. Conclusion: The expression of bcl-2 and p53 represent biological characteristics of colorectal carcinomas. Assessment of both bcl-2 and p53 status may be valuable in predicting the prognosis of patients.

Objective: This study is aimed at developing a simple and easy way to generate dendritic cells (DCs) from human peripheral blood monocytes (PBMCs) in vitro. Methods: PBMCs were isolated directly from white blood cell rather than whole blood and purified by patching methods (collecting the attached cell and removing the suspension cell). DCs were then generated by culturing PBMCs for six days with 30 ng/ml recombinant human granulocyte-macrophage stimulating factor (rhGM-CSF) and 20 ng/ml recombinant human interleukin-4 (rhIL-4) in vitro. On the sixth day, TNF-alpha (TNFα) 30 ng/ml was added into some DC cultures, which were then incubated for two additional days. The morphology was monitored by light microscopy and transmission electronic microscopy, and the phenotypes were determined by flow cytometry. Autologous mixed leukocyte reactions (MLR) were used to characterize DC function after TNFα or lipopolysaccharide (LPS) stimulations for 24 h. Results: After six days of culture, the monocytes developed significant dendritic morphology and a portion of cells expressed CD1a, CD80 and CD86, features of DCs. TNFα treatment induced DCs maturation and up-regulation of CD80, CD86 and CD83. Autologous MLR demonstrated that these DCs possess potent T-cell stimulatory capacity. Conclusion: This study developed a simple and easy way to generate DCs from PBMCs exposed to rhGM-CSF and rhIL-4. The DCs produced by this method acquired morphologic and antigenic characteristics of DCs.

Objective: To study the clinical features of chronic hepatitis B (CHB) patients with tyrosine-methionine-aspartate-aspartate (YMDD) mutation after lamivudine therapy. Methods: This investigation was a retrospective study of 63 CHB patients with YMDD mutation during lamivudine therapy. Clinical data, including period and types of YMDD mutation; hepatitis B virus (HBV) DNA levels and alanine aminotransferase (ALT) levels before and after YMDD mutation were measured. YMDD mutation in the HBV DNA polymerase gene was determined using polymerase chain reaction (PCR) and direct sequencing. HBV DNA quantification was determined using real-time PCR. Relevant serum markers of HBV were measured. The follow-up period was 12 months after YMDD mutation. Results: YMDD mutation occurred 7~44 months (median, 21.5 months) after the start of lamivudine therapy. The majority of the cases (42/63, 66.6%) had YMDD mutants detected between 12 and 24 months. Four types of YMDD mutation were observed in this study, rtL180M/M204V mutation was the predominant type (26/63, 41.3%). A proportion of patients (16/63, 25.4%; 12/63, 19.1%) had higher HBV DNA levels and ALT levels (after mutation vs before mutation), respectively. Conclusion: The majority of patients with YMDD mutants had similar or lower HBV DNA levels and ALT levels compared with baseline values. This subset of patients might have benefited from the continued lamivudine therapy. The patients with increased ALT and HBV DNA levels (breakthrough hepatitis) should benefit from the addition of a newer nucleotide analogue (e.g. adefovir).

Objective: Acute pulmonary thromboembolism (PTE) is a serious high mortality pulmonary vascular disease whose effective treatment decreases morbidity and mortality. To determine if low-molecular-weight-heparin (LMWH) is clinically as efficient and safe as unfractionated heparin (UH) in patients with diagnosis of acute non-massive PTE, our study compares the efficacy, adverse effects and costs of LMWH and UH. Methods: One hundred and fourteen patients with non-massive acute PTE were randomly divided into LMWH (nadroparin calcium) and UH groups. Oxygenation index, D-dimer, fibrinogen (FG), lung ventilation/perfusion (V/Q) scan and computed tomography pulmonary angiography (CTPA) were observed before anticoagulation and on day 14 after anticoagulation. Results: In both groups, the ABG (arterial blood gas) analysis showed PaO₂ and PaCO₂ were elevated, P(A-a)O₂ was decreased and oxygenation index (PaO₂/FIO₂) was elevated, D-dimer and fibrinogen were decreased, lung V/Q and CTPA showed embolized segments reduced (P<0.05). Hemorrhage and thrombocytopenia occurred in 3.5% of the LMWH group. Hemorrhage occurred in 5.3% and thrombocytopenia occurred in 7.0% of the UH group. The average cost in the LMWH group was RMB 1218.60 Yuan and RMB 1541.40 Yuan in the UH group. Conclusion: LMWH and UH are equally effective for treatment of non-massive acute PTE, but LMWH may have a lower prevalence of complications and is less expensive.

Objective: We aim to describe the environment iodine concentration in salt, water and soil along Zhejiang Province coast in the China foreland. It will be helpful for us to judge whether this area is insufficient in iodine and universal iodized salt is necessary or not. Methods: We collected iodized salt samples, drinking water samples (tap water in the towns, and well water or spring water in the villages), water samples from different sources (ditches, lakes, rivers) and soil samples through random sampling in June, 2005. Salt, water and soil iodine was detected by arsenic-cerium redox method. Statistical analysis was expressed as mean±SEM by Windows SPSS 13.0. Results: (1) The iodine concentration in salt was 27.9±4.33 mg/kg (n=108). (2) Seventy-five water samples were collected. The water iodine value was 0.6~84.8 μg/L (mean of 11.66 μg/L). The watershed along the Qiantang River has significantly higher iodine content than the water in Lin’an in mountain area (P<0.01). The iodine content and mean iodine content of tap water, well or spring water and natural water sources were 4.30±2.43 μg/L (n=34), 23.59±27.74 μg/L (n=19) and 12.72±10.72 μg/L (n=22) respectively. This indicated that among environmental water sources, the ditch iodine content was the highest with river water iodine being the lowest (P<0.01). (3) Soil iodine value was 0.11~2.93 mg/kg (mean of 1.32 mg/kg). Though there was no statistical difference of soil iodine in different districts (P=0.131), soil iodine content correlated positively with water iodine content. Conclusion: Iodine concentration in salt accords with national policy of adding iodine in salt. Foreland has more iodine in water than mountain area. The data reflected that water and soil iodine in foreland area was not high, which suggests universal iodized salt should be necessary. Environment iodine has relatively close association with pollution.

A novel technique of three-dimensional (3D) reconstruction, segmentation, display and analysis of series slices of images including microscopic wide field optical sectioning by deconvolution method, cryo-electron microscope slices by Fourier-Bessel synthesis and electron tomography (ET), and a series of computed tomography (CT) was developed to perform simultaneous measurement on the structure and function of biomedical samples. The paper presents the 3D reconstruction segmentation display and analysis results of pollen spore, chaperonin, virus, head, cervical bone, tibia and carpus. At the same time, it also puts forward some potential applications of the new technique in the biomedical realm.

To investigate the features of electroencephalography (EEG) power and coherence at rest and during a working memory task of patients with mild cognitive impairment (MCI). Thirty-five patients (17 males, 18 females; 52~71 years old) and 34 sex- and age-matched controls (17 males, 17 females; 51~63 years old) were recruited in the present study. Mini-Mental State Examination (MMSE) of 35 patients with MCI and 34 normal controls revealed that the scores of MCI patients did not differ significantly from those of normal controls (P>0.05). Then, EEGs at rest and during working memory task with three levels of working memory load were recorded. The EEG power was computed over 10 channels: right and left frontal (F3, F4), central (C3, C4), parietal (P3, P4), temporal (T5, T6) and occipital (O1, O2); inter-hemispheric coherences were computed from five electrode pairs of F3-F4, C3-C4, P3-P4, T5-T6 and O1-O2 for delta (1.0~3.5 Hz), theta (4.0~7.5 Hz), alpha-1 (8.0~10.0 Hz), alpha-2 (10.5 ~13.0 Hz), beta-1 (13.5~18.0 Hz) and beta-2 (18.5~30.0 Hz) frequency bands. All values of the EEG power of MCI patients were found to be higher than those of normal controls at rest and during working memory tasks. Furthermore, the values of EEG power in the theta, alpha-1, alpha-2 and beta-1 bands of patients with MCI were significantly high (P<0.05) in comparison with those of normal controls. Correlation analysis indicated a significant negative correlation between the EEG powers and MMSE scores. In addition, during working memory tasks, the EEG coherences in all bands were significantly higher in the MCI group in comparison with those in the control group (P<0.05). However, there was no significant difference in EEG coherences between two groups at rest. These findings comprise evidence that MCI patients have higher EEG power at rest, and higher EEG power and coherence during working conditions. It suggests that MCI may be associated with compensatory processes at rest and during working memory tasks. Moreover, failure of normal cortical connections may be exist in MCI patients.

To evaluate the phacoemulsification and intraocular lens implantation in patients with sensory exotropia subsequent to senile cataract. The authors prospectively studied the role of phacoemulsification and intraocular lens implantation on 25 patients by observing visual acuity, ocular alignment, binocular vision and diplopia pre-, 1 month post- and 3 months post-operation. The patients underwent follow-up for three months. Postoperatively, one patient had a corrected visual acuity of 20/50, and 24 patients had 20/40 or better. The ocular alignment, binocular vision and diplopia were resolved spontaneously. Phacoemulsification and intraocular lens implantation performed together is effective on sensory exotropia subsequent to senile cataract.

Objective: This study aimed to establish chemiluminescent immunoassay (CLIA) for quantitative determination of theophylline levels in human serum. Methods: To measure the concentration of theophylline (n=122) and evaluate the assay. Results: The linear range of the CLIA method was 0.51~40 mg/L (Y=1.02X+0.44, r=0.995). The intra and inter CV (coefficient variance) of CLIA were 3.20% and 3.57%, respectively. The average recovery rate was 102.3%. This method was free from interference by brilirubin (<200 μmol/L), hemoglobin (<10 g/L), and triglycerides (<15 mmol/L). Conclusion: This method is simple, convenient and precise for clinical pharmacokinetics study of theophylline.

We identified a novel gene ST13 from a subtractive cDNA library of normal intestinal mucosa in 1993, more studies showed that ST13 was a co-chaperone of Hsp70s. Recently we detected the ST13 gene expression in tumor tissue and adjacent normal tissue of the same colorectal cancer patient and investigated if the ST13 gene expression might have any prognostic value. Analysis was performed at molecular level by reverse transcription-PCR using real-time detection method. We measured two genes simultaneously, ST13 as the target gene and glyceraldehydes-3-phosphate dehydrogenase as a reference gene, in primary colorectal tumor specimens and tumor-adjacent normal mucosa specimens from 50 colorectal cancer patients. The expression levels of the ST13 gene were significantly decreased in primary tumors compared with adjacent mucosa (P<0.05). But there were no significant differences in the expression of ST13 as compared with different Dukes’ stage, tumor differentiation grade, invasion depth, lymph node metastasis and disease-specific survival.

Objective: To study the effect and implication of nonmyeloablative donor specific bone marrow (DSBM) infusion on the immunoreaction of liver allotransplantation. Methods: Orthotopic liver transplantation model was used in this study. Groups were set as follows: Group I, syngeneic control (Wistar-to-Wistar); Group II, acute rejection (SD-to-Wistar); Group III, acute rejection treated with cyclosporine A (CsA) by intramuscular injection (SD-to-Wistar+CsA); Group IV, bone marrow infusion at 7 d pretransplantation followed by short-term CsA treatment (SD-to-Wistar+DSBM); Another group of short-term CsA treatment preoperatively without bone marrow infusion was also set as control. General characteristics and survival time were observed. Histological grades of rejection were determined by pathological examination. IL-2 and IFN-γ level in peripheral blood and donor liver were detected respectively by Enzyme-Linked Immuno-Sorbent Assay (ELISA) and Western blot. Chimerism of donor cells was measured by PCR for a male-specific marker (Y-chromosome-specific sequence, Sry). Results: No signs of rejection were found in Group I. Acute rejection occurred in both Group II and the short-term CsA treated group. All the recipients died at (9~15) d posttransplantation with a median survival time of (10.7±0.5) d and (11.2±2.4) d, respectively. Only mild rejection could be seen in Group III. In Group IV, 4 out of 6 recipients had long-term survival (>100 d), the histological grade of rejection was significantly lower than that of Group II, so did the expression level of IL-2 and IFN-γ in both peripheral blood and grafted liver. Y-chromosome-specific sequence (Sry) of male SD rats could be detected in the bone marrow, spleen and thymus of female recipients at 15 d after bone marrow infusion. Conclusion: Mild preconditioning nonmyeloablative donor specific bone marrow infusion can enhance chimerism formation in recipients, alleviate the rejection of liver allotransplantation and prolong survival of liver allotransplantation.

Objective: To study the clinical therapy and prognosis in children with transient congenital hypothyroidism (CH). Methods: Fifty-seven children with CH diagnosed after neonatal screening were treated with low-dosage levothyroxine (L-T4). Follow-up evaluation included the determination of TT3, TT4 and TSH serum levels and the assessment of thyroid gland morphology, bone age, growth development and development quotients (DQ). A full check-up was performed at age 2, when the affected children first discontinued the L-T4 treatment for 1 month, and one year later. Development quotients were compared with a control group of 29 healthy peers. Results: The initial L-T4 dosage administered was 3.21~5.81 μg/(kg·d) with an average of (16.25±3.87) μg/d. Mean duration of therapy was (28.09±9.56) months. No significant difference was found between study group and control group in the DQ test (average score (106.58±14.40) vs (102.4±8.6), P>0.05) and 96.49% of the CH children achieved a test score above 85. Bone age, 99mTc scans and ultrasonographic findings were all normal, and evaluation of physical development was normal too, as were the serum levels of TT3, TT4 and TSH after one year of follow-up. Conclusion: A L-T4 dosage of 3.21~5.81 μg/(kg·d) was found sufficient for the treatment of transient CH. The treated children showed satisfactory overall mental and physical development at age 2. So it is possible for CH children to stop taking medicine if their laboratory findings and physical development are all normal after regular treatment and 2~3 years of follow-up.