Objective To evaluate the effect of open reduction and internal fixation in treatment of calcaneal fractures in aged patients. Methods Between January 1997 and June 2007,37 patients (41 fractures)at a mean age of68.3 years(60-78 years)with calcaneal fractures underwent surgical operations.According to Sanders classification based on CT scanning.14 fractures were rated as typeⅡ,18 as type Ⅲ and 9 as type Ⅳ.Of all,5 fractures were treated with reconstruction plate,18 with shapeable titanic plate,4 with Y-shaped plate,12 Depuy titanic plate and 2 with AO locking titanic plate plus open reduction and internal fixation.The foot function was evaluated by X-ray,American Orthopedic Foot and Ankle Society(AOFAS)Score and Maryland Foot Score. Results A total of 27 patients (60.9%)with 31 fractures were followed up for 12-48 months(average 27.6 months),which showed that all fractures were healed,with morphous improvement of the calcanus determined by 8 parameters measured in X-ray films.The average active range of motion was 36°of plantar flexion,12°of dorsiflexion,15°of inversion,and 10°of eversion.According to Maryland Foot Score,functional foot score was excellent(90-100 points)in 16 fractures,good(75-89 points)in 13 and fair(50-74 points)in 2,with excellence rate of 94%(29/31). Conclusion Open reduction and intemal fixation can gain good clinical result in severe calcaneal fractures in aged patients with obvious dislocation of posterior articular facet.

ObjectiveTo evaluate the clinical outcome of new proximal femoral nail antirotation (PFNA) in treatment of subtrochanteric fractures. MethodsThere were 31 patients with traumatic subtrochanteric fractures intramedullarily fixed with PFNA. The patients included 14 males and 17 fe-males at a mean age of 54.6 years (range 34-90 years). According to Seinsheimer classification of sub-trochanteric fractures, 14 patients were with type Ⅱ fractures (including four patients with type ⅡA,seven with type ⅡB and three with type ⅡC), 12 with type Ⅲ fractures (including eight with type ⅢA and four with type ⅢB), two with type Ⅳ fractures and three with type Ⅴ fractures. Close reduction was performed under traction and C-arm fluoroscopy in 26 patients and open reduction through a mini-incision before inserting the nail in five patients due to difficult close reduction. ResultsThe operation lasted for a mean time of 72.5 minutes (range 45-120 minutes), with mean blood loss of 127.5 ml (range 100-350 ml). Of all, 27 patients (87.1%, 27/31) were followed up for a mean duration of 14.3 months (range 8-24 months), which showed fracture union in all patients, with a mean union period of 17.4 weeks (range 10-21 weeks). According to Harris Hip Rate Scale, the results were. excellent in 21 pa-tients, good in two and fair in four, with excellence rate of 85.2% (23/27). ConclusionsPFNA is an effective device for treatment of subtrochanteric fracture, with a high union rate and a low complication rate, for it has advantages of easy use and minor trauma, low bleeding and stable fixation in operation.

OBJECTIVETo analyze the constituent proteins in donkey hide, the key ingredient for Ejiao, an important traditional Chinese medicine for the blood-related conditions, in hope to eventually decipher the biochemical mechanism behind Ejiao's prominent medicinal efficacy.

METHODTwo methods were employed to extract proteins in donkey skin. One used TriPure isolation reagent to extract the total proteins in donkey skin. Another used 1% sodium dodecyl sulfate (SDS) to heat the sample at 100 degrees C overnight. And then sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and capillary HPLC were used to analyze the component of proteins.

RESULTThere are not only collagen alpha1 (I) and collagen alpha2 (I), but also serum albumin in donkey skin. The content is over 25% in total proteins with the method of TriPure isolation reagent. The content of donkey serum albumin is up to 20% with the method of 1% SDS heating. And two bands, molecular weight are nearly 200 kDa,were found on 7.5% SDS-PAGE. Extracted these proteins to analyze with capillary HPLC, they were found to be the complex products of collagen and serum albumin of donkey.

CONCLUSIONDonkey serum albumin is a main protein component in the hide, which is a clue to expose is the effect of Ejiao on blood.

Animals ; Chromatography, High Pressure Liquid ; Collagen Type I ; analysis ; chemistry ; metabolism ; Collagen Type II ; analysis ; chemistry ; metabolism ; Drug Interactions ; Electrophoresis, Polyacrylamide Gel ; Equidae ; Molecular Weight ; Protein Binding ; Serum Albumin ; analysis ; chemistry ; metabolism ; Skin ; chemistry

Resistance to chemotherapy is a major obstacle for the effective treatment of advanced ovarian cancer. The mechanism of chemoresistance is still poorly understood. Recently, more and more evidence showed microRNAs (miRNAs) modulated many key molecules and pathways involved in chemotherapy. microRNA-106a (miR-106a) has been implicated in many cancers, but its role in ovarian cancer and drug resistance still remains unexplored. This study was to investigate whether miR-106a mediated resistance of the ovarian cancer cell line A2780 to the chemotherapeutic agent cisplatin (DDP). The different levels of miR-106a in A2780 cells and their resistant variant A2780/DDP cells were identified by using real-time PCR. MTT assay and flow cytometry were used to analyze the effect of miR-106a on cisplatin resistance of these paired cells. Real-time PCR, Western blotting and luciferase reporter assay were applied to explore whether Mcl-1 was a target of miR-106a. As compared to A2780 cells, the expression of miR-106a was down-regulated in the cisplatin resistant cell line A2780/DDP. Moreover, knockdown of miR-106a dramatically decreased antiproliferative effects and apoptosis induced by cisplatin in A2780 cells, while overexpression of miR-106a significantly increased antiproliferative effects and apoptosis induced by cisplatin in A2780/DDP cells. Furthermore, miR-106a inhibited cell survival and cisplatin resistance through downregulating the expression of Mcl-1. Mcl-1 was a direct target of miR-106a. These results suggest that miR-106a may provide a novel mechanism for understanding cisplatin resistance in ovarian cancer by modulating Mcl-1.
Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Drug Resistance, Neoplasm ; genetics ; Female ; Humans ; MicroRNAs ; genetics ; Myeloid Cell Leukemia Sequence 1 Protein ; genetics ; Ovarian Neoplasms ; drug therapy ; genetics

Ovarian cancer is the fifth lethal gynecologic malignancy. Metastasis-associated gene 1 (MTA1) is overexpressed in many malignant tumors with high metastatic potential. This study investigated whether down-regulation of MTA1 expression by RNAi in A2780 ovarian cancer cells could affect proliferation, anoikis, migration, invasion and adhesion of the cells and to research the potential for MTA1 gene therapy of ovarian cancer. After transfection with effective Mta1 gene siRNA, the effects on proliferation, anoikis, migration, invasion and adhesion of A2780 cells were tested by MTT assay, flow cytometry, wound-healing assay, Transwell assay and adhesion assay. Expression levels of PTEN, beta 1 integrin, MMP-9, phosphor-AKT (Ser473), and total AKT activity were evaluated in control and transfected cells. The results showed that inhibition of MTA1 mediated by Mta1-siRNA transfection decreased the cell invasion, migration and adhesion, and induced the increased cell anoikis, but no significant difference was found in proliferation of A2780 cancer cells. In addition, beta 1 integrin, MMP-9, and phosphor-AKT protein levels were significantly down-regulated, while PTEN was significantly up-regulated. These results demonstrated that MTA1 played an important role in the cell metastasis in ovarian cancer. MTA1 could serve as another novel potential therapeutic target in ovarian cancer.
Apoptosis ; genetics ; Carcinoma ; genetics ; pathology ; secondary ; Cell Line, Tumor ; Cell Survival ; genetics ; Female ; Gene Targeting ; methods ; Genetic Therapy ; methods ; Histone Deacetylases ; genetics ; Humans ; Ovarian Neoplasms ; genetics ; pathology ; therapy ; RNA, Small Interfering ; genetics ; therapeutic use ; Repressor Proteins ; genetics ; Treatment Outcome

OBJECTIVETo observe the effect of paiqian chewing tablet (PQCT) on lead discharging and health in children.

METHODSAdopting self-control and inter-group control method, 94 children with blood lead level exceeding 100 microg/L were randomly divided into the observed group and the control group. The observation period for both groups was 30 days.

RESULTSAt the 20th and 30th day of treatment, the urinary lead output in the observed group was significantly higher than that in the control group (P < 0.05, P < 0.01), and showed significant difference as compared with that before treatment (P < 0.05). Besides, the total amount of urinary lead discharging in the observed group was significantly more than that in the control group (P < 0.05).

CONCLUSIONPQCT has markedly lead discharging improvement action with no influence on urinary calcium and zinc excretion. As all the routine indexes of blood and urine ranged within the normal extent, it demonstrated that PQCT was harmless to the health of observed individual.

Child ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Lead ; blood ; urine ; Lead Poisoning ; drug therapy ; Male ; Phytotherapy ; Tablets

Objective To detect the expressions of DNA damage checkpoint genes including A TR, A TM, Chk1 and Chk2 in human primary gliomns and explore their relations with tumor progression. Methods SYBRTM Green real-time quantitative PCR was performed to detect the expressions of ATR, A TM, Chk1 and Chk2 genes in 35 cases of primary gliomas and 10 of normal brain tissues. Results In glioma tissues of various pathological grades, the expressions of the target genes, with the exception of A TM gene, were significantly increased as compared to those in normal brain tissues (P<0.05). Chk1 gene expression was significantly higher in grade Ⅳ than in grade Ⅱ and Ⅲ gliomas (P<0.05), but no significant differences were found in A TR or Chk2 gene expression between grade Ⅱ, Ⅲ and Ⅳ gliomas (P>0.05). Conclusion The up-regulation of ATR, Chk1 and Chk2 genes in primary glioma suggests their association with the pathogenesis of glioma. Chk1 expression may indicate the malignancy of glioma and help evaluate the pathological grade of glioma.

This study was purposed to screen the drugs for regulating tissue factor (TF) gene expression through establishing stable cell line with luciferase gene having TF promoter transcription activity, so as to provide the basis for further studying the molecular mechanism of screened drugs. A series of luciferase reporter gene plasmids under control of 5'-truncated TF promoter (including -2174 bp - +128 bp, -684 bp - +128 bp, -247 bp - +128 bp and -201 bp - +128 bp) were constructed. The above plasmids were separately electroporated into U937 cells to establish stably transfected sublines. The function of stable cell line was testified by treatment with ATRA, the luciferase gene activity was analyzed by treating established cell line with bortezomib (BTZ) and CDA-II, and drugs for regulating TF gene expression were screened. The results indicated that the BTZ of 5 nmol/L could activate TF gene transcription activity, up-regulate the expression level of TF transcripts; CDA-II of 1 mg/ml could suppress TF gene transcription activity, down-regulate the expression level of TF transcripts. The functional analysis of TF promoter transcription revealed that the region of regulating TF promoter transcription activity by BTZ and CDA-II was between -201 to 0 bp. It is concluded that stable cell line U937 expressing luciferase activity of TF promoters is established, the novel drugs regulating TF gene expression are screened out by means of this established cell line. This study provides basis for screening the new drugs and further studying their molecular mechanisms.
Antineoplastic Agents ; pharmacology ; Boronic Acids ; pharmacology ; Bortezomib ; Drug Screening Assays, Antitumor ; Gene Expression ; Humans ; Molecular Sequence Data ; Promoter Regions, Genetic ; Pyrazines ; pharmacology ; Thromboplastin ; genetics ; Transcription Factors ; genetics ; Transcriptional Activation ; U937 Cells

OBJECTIVETo study the clinicopathologic features of follicular dendritic cell sarcoma (FDCS) and its differential diagnosis.

METHODSTen cases of FDCS were studied by light microscopy, immunohistochemistry and in-situ hybridization. The clinical features and follow-up information were analyzed.

RESULTSAmongst the 10 cases of FDCS studied, the male-to-female ratio was 1:1. The mean age of the patients was 42 years. Six of them were located in cervical and peritoneal lymph nodes and four in extranodal sites (including tonsil, pelvic cavity, tail of pancreas and spleen). Histologically, the tumor cells had whorled, storiform or diffuse growth patterns. They were spindle in shape and contained syncytial eosinophilic cytoplasm, with round or oval nuclei, vesicular chromatin, distinct nucleoli and a variable number of mitotic figures. Multinucleated tumor giant cells and intranuclear pseudoinclusions were occasionally seen. There was a sprinkling of small lymphocytes and neutrophils within the tumor as well as in the perivascular region. Immunohistochemical study showed that the tumor cells were diffusely or focally positive for CD21, CD23, CD35 and D2-40, but negative for LCA, CD20, CD3, CD1a, HMB45 and CK. Some of them showed EMA, CD68 and S-100 reactivity. In-situ hybridization for Epstein-Barr virus-encoded RNA (EBER) showed positive signals in only one case (which was diagnosed as inflammatory pseudotumor-like FDCS). Of the 7 patients with follow-up information available (duration: 2 months to 39 months; mean: 14 months), 2 cases with paraneoplastic pemphigus died of pulmonary infection at 5 and 7 months respectively. The remaining 5 patients were alive and disease-free after surgical excision (+/- chemotherapy and radiotherapy).

CONCLUSIONSFDCS is a rare low to intermediate-grade malignant tumor. Appropriate application of FDC markers, such as CD21, CD35 and D2-40, would be helpful for arriving at a correct diagnosis. Most cases are associated with good prognosis after surgical treatment, with or without chemotherapy and radiotherapy. Patients with paraneoplastic pemphigus carry a less favorable prognosis.

Adult ; Antibodies, Monoclonal, Murine-Derived ; metabolism ; Dendritic Cell Sarcoma, Follicular ; complications ; metabolism ; pathology ; surgery ; Dendritic Cell Sarcoma, Interdigitating ; pathology ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Lymph Node Excision ; Lymph Nodes ; pathology ; surgery ; Male ; Meningioma ; pathology ; Middle Aged ; Nasopharyngeal Neoplasms ; pathology ; Paraneoplastic Syndromes ; complications ; Pemphigus ; complications ; Receptors, Complement 3b ; metabolism ; Receptors, Complement 3d ; metabolism ; Receptors, IgE ; metabolism ; Tonsillar Neoplasms ; metabolism ; pathology ; surgery ; Young Adult

This study was aimed to establish a stable subline of K562 cells (K562-HMGB1) overexpressing HMGB1 protein and K562-HMGB1 sublines served as control, so as to provide a basis for exploring the role of hmgb1 gene in occurrence and development of leukemia and their mechanism. Protein-coding gene of hmgb1 was amplified by PCR with cDNA as template, which was synthesized by reverse transcription from total RNA extracted from U937 cells. The PCR-amplified hmgb1 gene was ligated into PMD18-T vector (PMD18-T-HMGB1 vector), and then transformed into E. coli strain DH5α. DH5α containing PMD18-T-HMGB1 vector were grown on LB agar plate supplemented with 100 µg/ml ampicillin overnight. The single ampicillin-selected DH5α clone was picked for culturing overnight and then harvested for plasmid extraction. The extracted plasmid was characterized to contain hmgb1 gene digested with the desired restriction enzymes of KpnI/XhoI. The correctness of hmgb1 sequence was confirmed with DNA sequencing. The insert of hmgb1 gene contained in PMD18-T-HMGB1 vector was cut out with restriction enzymes of KpnI/XhoI and then ligated into eukaryotic expression vector pcDNA3.1 to form pcDNA3.1-HMGB1 vector. 10µg of pcDNA3.1-HMGB1 or pcDNA3.1 plasmid was separately electroporated into K562 cells. At 48 hours after electroporation the cells were cultured with G418 at a final concentration of 800 µg/ml for over 2 weeks. Finally stably transfected sublines of K562 cells containing hmgb1 gene (K562-HMGB1), and of K562 containing pcDNA3.1 vector (K562-pcDNA3.1) served as a control, were obtained. The transcriptional or translational expression of hmgb1 gene was detected with RT-PCR or Western blot, respectively, to testify transfected efficiency and validity of stable subline of K562-HMGB1. The results indicated that the eukaryotic expression vector pcDNA3.1-HMGB1 plasmid was successfully constructed and was electroporated into K562 cells. The transcriptional or translational expression of hmgb1 gene in the stable subline of K562 cells containing hmgb1 gene was overexpressed. It indicated that stable subline of K562-HMGB1 cells was successfully established. It is concluded that the stable sublines of K562-HMGB1 cells or K562-pcDNA3.1 cells are successfully established, which provides a basis for exploring the roles and mechanisms of hmgb1 gene in leukemogenesis and development of leukemia.
Gene Expression ; Genes, Regulator ; Genetic Vectors ; HMGB1 Protein ; genetics ; Humans ; K562 Cells ; metabolism ; Plasmids ; Transformation, Genetic