Objective To explore the biological features of hematopoiesis reconstitution by using genetic marking.Methods NeoR gene was transduced into bone marrow(BM)cells of mouse mediated by liposome.Then these cells were transplanted into lethally irradiated recipients.The hematopoiesis reconstitution was observed and the marker gene in spleen and BM cells of recipients after hematopoiesis reconstitution was examined.Results The transplanted recipients remained alive and healthy.But the irradiated mice with no transplantation died from BM aplasia soon.Meanwhile,the cells from spleen and BM of transplanted mice could be alive in G418 system,and contained the DNA fragments of NeoR gene by PCR.Conclusion Gene-modified BM cells could be used to reconstitute hematopoiesis successfully and express the foreign gene to some extent stably.

Objective To investigate whether human monocyte-derived dendritic cells (DCs) can express p210 Bcr-Abl protein and induce antigen-specific CTL responses in vitro after transfection with total RNA of K562 cells (K562-RNA).Methods Immature DCs were derived from human peripheral blood monocytes after 5 day incubation in the presence of GM-CSF and IL-4. The cells were then transfected with K562-RNA using electroporation or DOTAP lipofection. To verify the successful transfection of DCs with K562-RNA, Bcr-Abl fusion gene expression was determined by RT-PCR and Western blot. The immune phenotypes of the DCs were analyzed by flow cytometry. CTL cytotoxicity was assayed by propidium iodide (PI) stain and flow cytometry. The amount of DCs, CD1a expression and purity of DCs were measured by FACS.Results Bcr-Abl fusion gene appeared in the DCs after transfection with K562 cell total RNA. But 24 hours later, the Bcr-Abl mRNA from the K562-RNA transfected DCs disappeared, while the cells were expressing p210 Bcr-Abl protein. The transfected DCs could significantly promote T lymphocytes to kill the target K562 cells. We found that PBMC can be induced to DC in culture medium containing human plasma, suggesting a potential for clinical application.Conclusion Human dendritic cells transfected by K562 total RNA can induce effective p210 Bcr-Abl protein-specific immune responses, which might broaden the spectrum of possible DC-based clinical applications.

In this paper, the question about automatic brightness control for x-ray imaging systems based on CCD camera is discussed, and the structure and principle of an auto brightness control loop are analyzed along with the working procedure of the x-ray imaging system. A kind of digital brightness controller about a typical device and the designing idea of the computer brightness intelligent control software is introduced.
Algorithms ; Image Processing, Computer-Assisted ; Radiation Protection ; Radiographic Image Enhancement ; instrumentation ; Radiography ; instrumentation ; X-Ray Intensifying Screens

Based on a survey of community health service organization in several cities, community health service model based on the family clinic was compared with state-owned community health service model, and status quo, advantages and problems of family community health service organization were analyzed. Furthermore, policies for the management of community health service organization based on the family clinic were put forward.
China ; Community Health Services ; methods ; organization & administration ; Data Collection ; Delivery of Health Care ; organization & administration ; Hospitals, Community ; organization & administration ; Humans

In order to explore whether the conventional use of 5-fluorouracil (5-Fu) had any toxic effects on trabecular meshwork cells, bovine trabecular meshwork cells were cultured in vitro and exposed to 5-Fu at different concentrations. The cellular morphology, ultrastructure, mortality and phagocytosis were studied under light microscopy, transmission electron microscopy and methods of Wright's stain. It was found that the toxic effects of 5-Fu on the cells were in a dose-dependent mode. 1×10-1 mg/ml of 5-Fu caused a large part of cells rounded up, while 1×10-3 mg/ml of the drug only a rough appearance of the cell surface. Exposure to 1×10-2 mg/ml of 5-Fu made mitochrone swollen and rough endoplasmic reticulum enlarged, with the cell mortality being 50.5 %. The latex microspheres engulfed in cytoplasm in cells receiving 1×10-1 and 1×10-2 mg/ml of 5-Fu were significantly decreased as compared with those in the control group (P<0.01). It was concluded that the safe concentration of 5-Fu on bovine trabecular meshwork cells was 1×10-3 mg/ml and the conventional dosage of 5-Fu in clinical practice would not cause injury to trabecular meshwork cells.

To study the influence of siRNA targeting survivin gene on the biological behavior of hepatocellular carcinoma (HCC), one pair of 21bp reverse repeated motifs of survivin target sequence with 9 spacers were synthesized and inserted into plasmid psilencer2. 1 to generate siRNA eukaryotic expression vector. After stable transfection into HepG2 cells, the biological behaviors of the survivin siRNA transfected HCC cells were observed. After the recombinant plasmid Psilence (+)-survivin was successfully constructed, survivin mRNA and protein expression inhibition ratio reached 73 % and 75 % respectively compared to control groups. Transfected cells with survivin siRNA demonstrated significantly inhibited cell growth and increased apoptosis. Subsequent study in nude mouse model demonstrated lower succeeding rate in cells transfected with survivin siRNA and slow growth rate. The results elucidated the siRNA targeting survivin gene could specially suppress its expression in HepG2 cells and mhibit tumor cells growth both in vivo and in vitro. This provides a theoretical basis to turn the drug resistance in tumor cells.

The implications of Survivin, CyclinD1, p21WAF1, Caspase-3 in the development, progression and prognosis in cervical cancer were investigated. By using immunohistochemical SP method, the expression of Survivin, CyclinD1, p21WAF1 , Caspase-3 was detected in 41 cases of cervical cancer, 17 cases of cervical intraepithelial neoplasia (CIN) and 10 cases of normal tissues, and their relation with pathological grade, clinical stage, metastasis and survival time was analyzed.The results showed that the positive expression rate of Survivin, CyclinD1 in cervical cancer was significantly higher than in CIN group and normal control group (P＜0.05). The median survival time in the patients with cervical cancer positive for Survivin and CyclinD1 was significantly shorter than in those with negative expression (P＜0.05). The expression of both Survivin and CyclinD1 was not related with tumor grade, clinical stage and metastasis (P＞0. 05). The positive expression rate of p21WAF1 , Caspase-3 in cervical ca rcer was significantly lower than in CIN group and normal control group (P＜0.05), and had a close relation with tumor grade (P＜0.05). The expression of Survivin in cervical cancer in cervical cancer was negatively associated with that of Caspase-3 (P＜0.01), but positively with that of CyclinD1 (P＜0.01). Cox Multivariate analysis revealed that Survivin was the independent prognostic indicator influencing the survival time of the patients with cervical cancer (P＜0.05). It was suggested that the high expression of Survivin or CyclinD1, and low expression of p21WAF1 or Caspase-3 was closely correlated with the development of cervical cancer. Survivin and CyclinD1 could be used as a useful indicator to predict the prognosis of cervical cancer.

Summary: In order to investigate the effect of small ubiquitin-like modifier-1 (SUMO-1) on the p53-induced HepG2 cell apoptosis, HepG2 cells were transfected by recombinant plasmids as pwtp53, pMDM2 and pSUMO-1 respectively. Western blot was employed to detect the protein expression of the transfected recombinant plasmids and the rate of apoptosis was measured by flow cytometry. The results showed that in cells transfected with pwtp53 and pwtp53+pSUMO-1, the apoptosis rate was (16.79±1.62) % and (18.15±1.36) % respectively, while transfected with pwtp53+pMDM2, the rate was decreased to (5.17±1.23) %. The apoptosis rate was (14.06±1.84) % in the cells transfected with pwtp53+pMDM2+pSUMO-1, significantly higher than that in the cells Transfected with pwtp53+pMDM2 (P<0.01). The apoptosis rates in the cells were all less than 2 % and had no significant difference among the groups. It was suggested that in the HepG2 cells, SUMO-1 can increase the apoptosis induced by wild-type p53 through binding to p53 protein, post-translational modification and inhibiting the p53 degradation by MDM2.

Summary: A total of 92 patients with benign prostatic hyperplasia (BPH) were subjected to modified Madigan prostatectomy (MPC) for a much satisfactory effect in open prostatectomy surgery. Exposing anterior prostatic urethra near the bladder neck and conjunct cystotomy modified the MPC procedure. This modified procedure preserved prostatic urethra intact and could also deal with intracystic lesions at the same time. The intact of prostatic urethra was kept completely or largely in 86 cases. The amount of blood loss during modified procedure was less. The mean operative time was 105 min. Seventy patients had been followed up for 3-24 months. The postoperative average Qmax was 19.2 ml/s. The cystourethrography revealed that the urethra and bladder neck were intact in 10 patients postoperatively. Furthermore, the prostatic urethra was obviously wider after modified MPC. The modified MPC can reduce the occurrence of urethra injury and enlarge the MPC indications. The modified technique is easy to perform with less complications and much satisfactory clinical result.

In order to explore whether the conventional use of 5-fluorouracil (5-Fu) had any toxic effects on trabecular meshwork cells, bovine trabecular meshwork cells were cultured in vitro and exposed to 5-Fu at different concentrations. The cellular morphology, ultrastructure, mortality and phagocytosis were studied under light microscopy, transmission electron microscopy and methods of Wright's stain. It was found that the toxic effects of 5-Fu on the cells were in a dose-dependent mode. 1×10-1 mg/ml of 5-Fu caused a large part of cells rounded up, while 1×10-3 mg/ml of the drug only a rough appearance of the cell surface. Exposure to 1×10-2 mg/ml of 5-Fu made mitochrone swollen and rough endoplasmic reticulum enlarged, with the cell mortality being 50.5 %. The latex microspheres engulfed in cytoplasm in cells receiving 1×10-1 and 1×10-2 mg/ml of 5-Fu were significantly decreased as compared with those in the control group (P<0.01). It was concluded that the safe concentration of 5-Fu on bovine trabecular meshwork cells was 1×10-3 mg/ml and the conventional dosage of 5-Fu in clinical practice would not cause injury to trabecular meshwork cells.