Objective To investigate the psychological status and related factors in patients with mainte-nance hemodialysis in Huzhou city.Methods 376 cases of maintenance hemodialysis were selected as the research subjects in Huzhou First People's Hospital.The anxiety and depression status in maintenance hemodialysis patients were investigated by issuing questionnaires.To study the relationship between anxiety and depression,gender,age, social support,nutritional status,complications,and in maintenance hemodialysis patients.At the same time,the quality of life was evaluated by SF -36 scale.Results Issuing 376 questionnaires,a total of 373 points,the effective recovery rate was 99.20%.In 373 patients,there were 182 patients (48.79%)with no depression,mild depression in 78 cases (20.91%),moderate depression in 59 cases (15.82%),severe depression in 54 cases (14.48%). 51.21% patients had different degree of depression.Of the 373 patients,there were 123 patients (32.97%)with no anxiety,mild anxiety in 141 cases (37.80%),mild and moderate anxiety in 55 cases (14.75%),moderate anxiety in 33 cases (8.85%),severe anxiety in 21 cases (5.63%).A total of 67.03% patients had different degrees of anx-iety.Huzhou city maintenance hemodialysis,female patients had depression rate of 80.70%,anxiety rate of 65.50%, which were significantly higher than the rate of depression 55.45%,anxiety rate 39.11% in male patients,the differ-ences were statistically significant (P <0.01 -0.05).In maintenance hemodialysis patients,the anxiety and depres-sion status gradually increased with age.Severe malnutrition in maintenance hemodialysis patients,the rate of depres-sion,anxiety rates were 80.65%,90.32%,the rates of depression,anxiety were 44.49%,61.81% in normal and mild malnutrition patients,the differences were statistically significant (all P <0.01).SF -36 scale showed that most of the patients with maintenance hemodialysis patients had poor quality of life,and the function was limited. Conclusion Maintenance hemodialysis patients,the general existence of anxiety and depression in the mental state in Huzhou city.Moreover,the psychological state of the patients is influenced by sex,age,nutrition status,and other factors.In clinical,it is needed to provide effective psychological intervention for patients to improve the quality of life,according to the psychological status and related factors of maintenance hemodialysis patients.

BACKGROUND: The skill to culture osteoblasts primarily has been well developed. However, trypsinase can affect membrane protein of osteoblasts if the time of digestion is long. Therefore, it is of great significance to select an ideal method to avoid the damage from trypsinase to cells as possible when culturing osteoblasts.OBJECTIVE: To explore a novel method to isolate and culture SD rat osteoblasts in vitro, and identify the functions of the cells.DESIGN: Observational study.SETTING: Department of Orthopaedics, First Affiliated Hospital of Jinan University.MATERIALS: This experiment was carried out in the Department of Orthopaedics, First Affiliated Hospital of Jinan University from March to May in 2007. Eight SPF 24-hour old SD rats were used in the experiment. The rats, irrespective of gender, were provided by the Experimental Animal Center of Nanfang Medical University. The experimental animals were disposed according to ethical criteria. The main reagents were detailed as follows: collagenase Ⅱ (Sigma Company);trypsin (Sigma Company); alkaline phosphatase (ALP) kit (Nanjing Jiancheng Biological Products Company); SABC-1021(Wuhan Boster Biotechnology Company).METHODS: 24-hour old SD rats were chosen for experiment. The newly born SD rats were sacrificed by anesthesia and the cranial bones of the rats were obtained cleanly, erased completely of the periosteum and cut to blocks of I mm3. The cranial bones were digested by 0.25 % trypsinase for 20 minutes, then by 0. 1% type Ⅱ collagenase for 60 minutes. The digestive time of trypsinase was controlled in the process of digestion to avoid to harm the cells. The liquid was gathered and centrifuged. The cells were cultured in culture flask and were purified by many times adhered.MAIN OUTCOME MEASURES: Morphology observations under the inverted phase contrast microscope, transmission electron microscope, and scanning electron microscope were performed. The phenotype, calcium tuberculation and the expression of alkaline phosphatase were studied with alizarin red staining and modified Gomori Ca-Co assays respectively.The cells were also evaluated with collage Ⅰ immunohistochemical staining.RESULTS: The cultured cells had active proliferation ability. Cells showed multi-angle or fusiform shape. Nucleus was immature and organell was plentiful. Therefore, they had typical morphological characters of osteoblasts. Moreover, they showed the osteoblastic phenotypes such as their synthesis of alkaline phosphatase, collage Ⅰ and formation of calcium tuberculations.CONCLUSION: The cells cultured by our modified enzymatic digestion method had typical morphological and biological characteristics of osteoblasts.

Aim To investigate the effects of dalbinol on proliferation and apoptosis of human colon cancer HCT1 16 cells and its mechanisms.Methods Anti-proliferative effect of dalbinol was evaluated by MTT assay.The morphological changes of apoptosis were observed by Hoechst33342 staining.Apoptotic rate and ROS generation were analyzed by flow cytometry.The related proteins of Wnt/β-catenin pathway and the ap-optosis-associated proteins expression were measured by Western blot.Results The growth of HCT1 16 treated with dalbinol was inhibited in a dose and time dependent manner with IC50 (4.8 ±0.53 ),(2.5 ± 0.43)and (0.6 ±0.22)μmol·L-1 at 24,48 and 72 h,respectively.Typical morphological changes of ap-optosis such as cell shrinkage,karyopyknosis and nu-clear condensation were observed by Hoechst33342 staining.Meanwhile,the apoptotic rate and intracellu-lar ROS generation of dalbinol were both increased dose-dependently. Western blot results showed that dalbinol could activate the expression of cleaved Caspase-3 and cleaved PARP by decreasing anti-apop-totic protein levels such as Bcl-2 and Mcl-1 and in-creasing pro-apoptotic protein levels such as Bax and Bim,which induced further apoptosis.Moreover,dal-binol can reduce the protein expression of the total and nuclear β-catenin,but not cytoplasmic β-catenin by suppressing the protein expression of Dvl-2 and GSK-3β(pS9 ),as well as its target proteins c-Myc and Sur-vivin.Conclusion dalbinol can induce apoptosis in colon cancer HCT1 16 cells by upregulating the intra-cellular ROS generation and suppressing Dvl/GSK-3β/β-catenin pathway.

Objective To investigate the alterations and phenotypes of dendritic cells, inflamma-tory monocytes and macrophages in immunocompetent mice during Pneumocystis murina ( P.murina) infec-tion for further analysis of the function of these cells during P.murina infection.Methods Wild type male C57BL/6 mice at age 6-8 weeks were randomly divided into two groups including the group with P.murina infection and the group receiving sham surgery.The mice without any intervention were used to set up the blank control group.The loads of P.murina strains in lung tissues of each mouse were quantified by TaqMan real-time fluorescence polymerase chain reaction after the infection.Histopathological examination was per-formed to evaluate the degree of inflammation in lung tissues.The numbers of dendritic cells, inflammatory monocytes and macrophages in lung tissues, peripheral blood and bone marrow samples, and the changes of inflammatory monocytes in spleen tissues were measured by flow cytometry analysis.The expression of major histocompatability complexⅡ(MHCⅡ), CX3C chemokine receptor 1 (CX3CR1) and CC chemokine re-ceptor 2 ( CCR2 ) by dendritic cells, inflammatory monocytes and macrophages in lung tissues during P.murina infection were analyzed by flow cytometry analysis.All of the data were collected one, two, three and four weeks after the corresponding treatments.Results The loads of P.murina strains in P.murina in-fected mice were elevated after two and three weeks infection, but decline at week 4 (P>0.05).Significant pathological changes including the alveolar destruction, inflammatory cell infiltration and thickened alveolar septum in mice with P.murina infection were observed under a microscope at week 3 and week 4.Compared to the sham surgery treatment group, the number of CD11c+CD11b+dendritic cells were increased in lung tissues, but decreased in blood samples during P.murina infection ( P<0.05) .The levels of inflammatory monocytes in blood samples fell at week 3 and then rose at week 4 during P.murina infection (P<0.05). No significant difference with the change of macrophages in mice was observed during P.murina infection ( P>0.05).The CD11c+CD11b+dendritic cells in lung tissues of mice with P.murina infection expressed high levels of MHCⅡand CX3CR1, and low levels of CCR2.The inflammatory monocytes in lung tissues of mice expressed high levels of CCR2, moderate levels of MHCⅡand low levels of CX3CR1 during P.murina in-fection.High levels of CX3CR1 and low levels of MHCⅡ and CCR2 were observed in macrophages from lung tissues of mice with P.murina infection.Conclusion Highly expressed CD11c+CD11b+dendritic cells and MHCⅡwere detected in lung tissues of mice during P.murina infection, indicating that CD11c+CD11b+dendritic cells were involved in the host defense against P.murina infection.

Objective To evaluate the diagnostic value of endoscopic ultrasonography (EUS), and explore the efficacy of endoscopic treatment in patients with esophageal submucosal tumor. Methods Sixty-eight patients with esophageal submucosal tumor were selected, and the tumor was derived from the muscularis mucosa and submucosa according to the common endoscope and endoscopic ultrasonography detection. Endoscopic mucosal resection (EMR) was applied to remove submucosal tumor with diameter less than 1.0 cm, endoscopic piecemeal mucosal resection (EPMR) or endoscopic submucosal dissection (ESD) was applied to remove submucosal tumor with diameter 1.1 - 1.5 cm, and ESD was applied to remove submucosal tumor bigger than 1.5 cm. Samples were examined by pathology after treatment. Results Tumors in all the patients were completely removed, and the tumor diameter was 0.6-2.3 cm. Forty-one cases were treated with EMR, 9 cases were treated with EPMR and 18 cases were treated with ESD. Four patients had intra-operative bleeding that was stopped by electrocoagulation hemostasis. No perforation occurred in all cases. Postoperative pathology revealed 43 cases had leiomyoma, 23 cases had interstitialoma, and 2 cases had lipoma. Patients were reviewed by gastroscope 3 months after operation. The white scars formed in all patients, and there was no residue or recurrence. Conclusions Different origin layers and property of esophageal submucosal tumor can be diagnosed accurately by EUS, and endoscopic therapy (EMR, EPMR and ESD) is an effective treatment for submucosal tumor from muscularis mucosa and submucosa. Endoscopic therapy is safe and effective. It provides sufficient pathological information.

BACKGROUND: The structure of nanometer chitosan-sodium/collagen (nano-CS/COL) is similar to that of the extracellular matrix (ECM) in the nanometer level. Whether this can promote the adhesion and growth of bone marrow mesenchymal stem cells (MSCs) and the calcification?OBJECTIVE: To investigate the in vitro histocompatibility of nano-CS/COL. DESIGN: Single sample observation.SETTING: Department of Orthopaedics, First Hospital, Jinan University. MATERIALS: This study was performed at the Experimental Center, First Hospital Affiliated to Jinan University between March 2007 and July 2007. Ten 4-week-old female SD rats, of SPF grade, weighing 200 g, were provided by the Guangdong Provincial Laboratory Center [Permission No. SCXK (yue) 2003-0002]. The protocol was carried out in accordance with animal ethics guidelines for the use and care of animals. Nano-CS/COL METHODS: Bone marrow MSCs were isolated from SD rats and cultured. Cell surface antigen was detected by loss cellanalyticalmethod.Nano-CS/COLscaffold waspreparedbypolyelectrolyte confocallaser-scanning microscopy. The well-grown cells of the third passage were co-cultured in vitro on the nano-CS/COL scaffold. Taking simple nano-CS/COL scaffold material as control, the histocompatibility of scaffold material and cells were comprehensively evaluated by cell adherence rate, growth curve, cell activity and cycle, and scanning electron microscope observation.MAIN OUTCOME MEASURES: ① Identification of cell surface antigen marker after isolation and culture of bone marrow MSCs. ②The histocompatibility of nano-CS/COL material and bone marrow MSCs 2, 4 and 8 days after nano-CS/COL material compounded with cells. ③Determination of adherence rate of cells to nano-CS/COL material. ? Cell circle and activity detected 5 days after nano-CS/COL material compounding with cells. RESULTS: ① Detection results of cell surface antigen marker: The expression of CD29, CD106, CD44, CD34 and CD45 was 90.86%, 73.38%, 82.61%, 0.76% and 0.60%, respectively. ②Histocompatibility of bone marrow MSCs and nano-CS/COL material: It was shown under the scanning electron microscope that nano-CS/COL scaffold presented porous three-dimensional structure, and different sizes of macropoles and interconnected small pores. The interval porosity determined by quality assay was 85%-90%, and aperture averaged 150 μm (range 50 - 300u m). Two days after bone marrow MSCs compounded to nano-CS/COL scaffold, bone marrow MSCs presented globular shape and were scattered; Four days later, bone marrow MSCs presented shuttle shape, extended and anchored on the surface of nano-CS/COL by pseudopods; Eight days later, bone marrow MSCs proliferated and fused each other, and they secreted a lot of extracellular matrix, then which covered most material particles. ③ The adherence rate of bone marrow MSCs to nano-CS/COL: Bone marrow MSCs and nano-CS/COL were co-cultured 2 and 6 hours separately. The adherence rate of bone marrow MSCs was higher to nano-CS/COL scaffold than to simple chitosan scaffold. ④ Comparison of cells and cell cycle between on nano-CS/COL scaffold and on the chitosan scaffold: On the nano-CS/COL scaffold, cell activity was 96.67%, cell cycle at G0-G1 was 90.81%, at G2-M was 0.52% and at S was 8.66%. G2/G1 was 1.81. On the simple chitosan scaffold, cell activity was 95.27%, cell cycle at G0-G1 was 87.14%, at G2-M was 9.69%, and at S was 4.16%. G2/G1 was 1.80.CONCLUSION: Nano-CS/COL scaffold can be used as tissue engineering biomaterials because bone marrow MSCs can well grow on it.

Objective To evaluate the inactivation effect of heat inactivation and three common chemical disinfectants on enterovirus group A 71 (EV-A71),provide basic data for disinfection work after laboratory activities in enterovirus secondary biosafety laboratory.Methods EV-A71 virus was treated at different temperatures and different disinfectant concentrations according to different time.The inactivated virus was inoculated into 96-well cell culture plates,and the cytopathic effect was observed and recorded.Finally,the virus titer of the virus treatment solution was calculated.Results After inactivation at 56 ℃ for 30 min,the inactivation logarithm of EV-A71 was more than 6,and inactivation at 65 ℃ for 30 rin,70 ℃ for 10 min,no infection was observed.Three chemical disinfectants could effectively inactivate a certain titer of EV-A71 at a certain concentration and treatment time.Conclusions The inactivation effect of EV-A71 virus is related to factors such as thermal inactivation temperature,chemical disinfectant concentration,and treatment time.Studying the inactivation curve of EV-A71 is beneficial to the selection of inactivation conditions in different inactivation environments.

We report a complete genomic sequence of rare isolates (minor genotype) of the SARS-CoV from SARS patients in Guangdong, China, where the first few cases emerged. The most striking discovery from the isolate is an extra 29-nucleotide sequence located at the nucleotide positions between 27,863 and 27,864 (referred to the complete sequence of BJ01) within an overlapped region composed of BGI-PUP5 (BGI-postulated uncharacterized protein 5) and BGI-PUP6 upstream of the N (nucleocapsid) protein. The discovery of this minor genotype, GD-Ins29, suggests a significant genetic event and differentiates it from the previously reported genotype, the dominant form among all sequenced SARS-CoV isolates. A 17-nt segment of this extra sequence is identical to a segment of the same size in two human mRNA sequences that may interfere with viral replication and transcription in the cytosol of the infected cells. It provides a new avenue for the exploration of the virus-host interaction in viral evolution, host pathogenesis, and vaccine development.
Base Sequence ; China ; Cluster Analysis ; Gene Components ; Genetic Variation ; Genome, Viral ; Genotype ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; Sequence Analysis, DNA ; Severe Acute Respiratory Syndrome ; genetics