OBJECTIVETo detect the expression of CD133 in human tongue squamous cell carcinoma Tea8113 cell line and observe proliferation ability of CD133 groups in vitro.

METHODSLimiting dilution assay was employed to observe the proliferating character of Tca8113 single cell in vitro. The ability of growing as cancer spheroids was observed with ultra-low attachment plates. The flow cytometry was used to detect the expression of putative tumor-initiating cell marker CD133 in human tongue squamous cell carcinoma Tca8113 cell line. The selective technique of immunomagnetic beads was applied to purify CD133 tumor cells, CD133 tumor cells were cultured and their ability of proliferation were observed in vitro.

RESULTSAfter 12 days, the result of single cell culture in vitro revealed that about 5.23% of cultured Tca8113 cells possessed the capacity of continue proliferation. The cells line fromed floating clusters with one week of passaging cells into non-adherent plates. Approximately 0.95% of cells in Tca8113 cell line expressed CD133. Compared with CD133- cells and control Tca8113 cells, CD133+ cells demonstrated increased proliferation capacity. The proportion of CD133 cells decreased in culture as days passed. The percentage of CD133+ cells decreased from 92.45% to 1.62% in twelve days' culture.

CONCLUSIONTumor stem cells have the character of heterogenity and lower proportion of CD133 but higher ability of proliferation, and the diferentiation in human tongue squamous cell carcinoma Tca8113 cell line in vitro, CD133 may be one of makers for tumor-initiating cell of human tongue squamous cell carcinoma Tca8113 cell line.

Carcinoma, Squamous Cell ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Humans ; Tongue Neoplasms

Previous studies demonstrated that interleukin-12 (IL-12) enhances the non-MHC-restricted cytotoxic activity of NK cells and facilitate specific allogeneic human cytotoxic T lymphocyte responses against fresh leukemia cells and cell lines. The Wilms' tumor gene, WT1 mRNA, has been used as a marker of minimal residual disease (MRD) for evaluating therapeutic efficacy of patients with leukemia or myelodysplastic syndrome (MDS). This study was aimed to investigate whether in vitro IL-12 can lower WT1 gene expression in peripheral blood monuclear cells (PBMNC) from patients with leukemia or MDS. PBMNC from these 30 patients and 5 healthy volunteers were cultured at 5 x 10(5) cells/ml alone with or without 100 units/ml of IL-12 for 3 days. WT1 mRNA was measured by competitive reverse transcription polymerase chain reaction (RT-PCR) since WT1 mRNA is considered as a marker of minimal residual disease (MRD) in leukemia and MDS. The results demonstrated that WT1 mRNA in PBMNC of 5 healthy volunteers was less than 10(3) copies/microg of total RNA. Following the 3-day IL-12 treatment, mean WT1 mRNA of PBMNC was reduced from 10(4.8) to 10(4.2) copies/microg of total RNA in 6 CML patients, from 10(5.4) to 10(4.8) copies/microg in 12 MDS patients and from 10(5.0) to 10(4.2) copies/microg in 5 AML patients in CR, but not reduced in 5 of 7 AML in non-CR. It is concluded that IL-12 significantly decrease the quantity of leukemia cells in PBMNC of most patients with MDS, CML and AML in CR. IL-12 may be of considerable benefit in the elimination of MRD in patients with hematological malignancies.
Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Interleukin-12 ; pharmacology ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; metabolism ; Neoplasm, Residual ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; WT1 Proteins ; biosynthesis ; genetics

AIMTo identify specifically expressed genes in the adult and fetal testes.

METHODSA human testis cDNA microarray was established. Then the mRNA of adult and fetal testis was purified and probes were prepared by a reverse transcription reaction with the testis mRNA as template. The microarray was hybridized with probes of adult and fetal testes. The nucleic sequences of differentially expressed genes were determined and homologies were searched in the databases of the GenBank.

RESULTSWhen hybridized with adult or fetal testis probes, the positive clones were 96.8 % and 95.4 %, respectively. Among these genes, one was a new testis-specific gene, which was named TSP1. TSP1 was highly expressed in human adult testis. The cDNA of TSP1 was 1,484 bp in length. The cDNA sequence of this clone was deposited in the Genbank (AF333098). TSP1 was also determined as Interim Gen Symbol (Unigene, No. Hs.98266). Protein analysis showed that TSP1 contained two functional domains: an N-terminal basic helix-loop-helix (bHLH) and a C-terminal leucine zipper (Zip). Homologous analysis showed that the 430 amino acid sequences deduced from the 1293 bp open reading frame (ORF) had a homology with the human gene FLJ2509 (AK098575). TSP1 had also a sequence homology with Spz 1 protein of mouse. Expression profiles showed that TSP1 was specifically and strongly expressed in the testis.

CONCLUSIONTSP1 is a gene highly expressed in adult testis. It may play an important role in spermatogenesis in the humans.

Adult ; Amino Acid Sequence ; genetics ; Base Sequence ; genetics ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; Fetus ; metabolism ; Gene Expression ; Genes ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Sequence Homology, Amino Acid ; Testis ; embryology ; metabolism ; Transcription Factors ; chemistry ; genetics ; metabolism

OBJECTIVETo investigate the prevalence and associated risk factors of bacterial translocation (BT) in patients with cirrhosis after liver transplantation and analyze the effect of BT on bacterial infection after the surgery.

METHODSMesenteric lymph nodes (MLN), portal vein blood, and peripheral blood were collected during the liver transplantation for microbiological culture from 78 patients with cirrhosis. And meanwhile, all related clinical data were analyzed to investigate the risk factors of BT and its relationship with post-liver transplantation infections.

RESULTSBT was occurred in 8 of 78 cirrhotic patients (10.3%) and positive-rate of MLN culture was 5/8. Gram-negative aerobic bacillus was the main causative bacterium of BT (5/9), followed by Gram-positive aerobic enterococcus (22.2%, 2/9). Total bilirubin level in patients with BT was significantly higher than that in patients without BT.

CONCLUSIONSIt suggests that hyperbilirubinemia is the only risk factor for BT, and BT is associated with an increased infectious rate after liver transplantation.

Adult ; Bacterial Infections ; blood ; etiology ; Bacterial Translocation ; Female ; Humans ; Intestines ; microbiology ; Liver Cirrhosis ; microbiology ; surgery ; Liver Transplantation ; adverse effects ; Male ; Middle Aged ; Peritonitis ; etiology ; Postoperative Complications ; etiology ; microbiology ; Risk Factors

OBJECTIVETo investigate the approach of open reduction and internal fixation for the treatment of anteromedial coronoid facet fractures and associated injury.

METHODSFrom July 2009 to August 2011, 6 coronoid anteromedial facet fractures were treated (4 males and 2 females,the average age was 32.6 years old,ranged from 19 to 49 years old) in our hospital. Three patients had fractures in the left side and 3 in the right(4 dominant sides and 2 non dominant sides). All the patients had close fractures. All the fractures were subtype 2 or subtype 3 of coronoid anteromedial facet fracture according to O'Driscoll classification. The anteromedial incision of elbow and the approach of splitting flexor digitorum superficialis between flexor carpi radialis and palmaris longus were used to apply internal fixation with mini-plate. After fixation of coronoid fracture, widening of radiohumeral joint interspace under varus stress occurred in 3 cases,elbow varus posteromedial rotational instability was showed,and the exploration and repair of radial collateral ligament was not carried out. Plaster was applied for 2 weeks before rehabilitation and the time of plaster application extended to 4 weeks in the patients showed varus posteromedial rotational instability. The motion degree,pain and stability, strength of elbow and hand were recorded,the elbows were evaluated with modified An and Morrey functional rating index.

RESULTSAll the patients were followed up,and the average duration was 9.3 months (ranged from 7.5 to 13 months). The strength of elbow and hand were equal to that in the contralateral side. The average flexion was (129.0+/-6.5) degree (ranged from 120 to 135 degree); the average extention was (4.0+/-4.2) degree (ranged from 0 to 10 degree); the average forearm pronation was (84.0+/-6.5) degree (ranged from 75 to 90 degree); average supination was (89.0+/-7.1) degree (ranged from 80 to 100 degree). One patient had ulnar sensory neuropathy after operation and healed in half-year, no postoperative complications as pain and instability accured. All the patients obtained excellent results according to modified An and Morrey functional rating index.

CONCLUSIONThe approach of splitting flexor digitorum superficialis between flexor carpi radialis and palmaris longus could be used in open reduction and internal fixation of anteromedial coronoid facet farcture, rehabilitation should be postponed in the patient showed elbow varus posteromedial rotational instability.

Adult ; Casts, Surgical ; Female ; Fracture Fixation, Internal ; adverse effects ; methods ; Humans ; Male ; Middle Aged ; Ulna Fractures ; surgery

OBJECTIVE: To analyze the relationship among the parameters by measuring the relevant parameters of the anteroposterior X-ray of both hips in patients after total hip arthroplasty, to discuss the reliable anatomical markers and reference standards of acetabulum placement in total hip arthroplasty, and finally to accurately control the abduction angle of acetabulum. METHODS: From January 2016 to June 2017, 282 patients (235 hips) underwent total hip arthroplasty and 128 patients(157 hips) met the inclusion criteria. There were 91 males and 37 females, 82 cases of the left hip and 75 cases of the right hip; ranging in age from 22 to 78 years old, with a mean of 55.1 years old. The abduction angle(β), ilium thickness (a), acetabular cup insertion depth (b), ischial thickness (c), acetabular cup insertion depth(d), acetabular abrasion and contusion depth(e) were measured on the postoperative AP X-ray of both hips, and the data were compared. RESULTS: There was a positive correlation between β and b (=0.424, =0.000), a negative correlation between β and d (=-0.407, =0.000), a positive correlation between β and b/a (=0.419, =0.000), a negative correlation between β and d/c (=-0.472, =0.000). There was a linear relationship between β and b/a (5.753, =0.000) and a linear relationship between β and d/c (-6.671, =0.000). CONCLUSIONS The outreach angle is mainly controlled by the distance between the outer edge of the cup and the outer edge of the cup in the inferior portion(d) during the operation. The distance b from the outer edge of the cup can be used as a reference.
Acetabulum ; Adult ; Aged ; Arthroplasty, Replacement, Hip ; Female ; Hip Prosthesis ; Humans ; Male ; Middle Aged ; Postoperative Period ; Radiography ; Young Adult

Objective: To investigate the application of single-molecule PCR (SM-PCR) in the detection of plasma ctDNA for the treat-ment of patients with advanced lung adenocarcinoma. Methods: In total, 30 patients diagnosed with advanced lung adenocarcinoma were enrolled between June 2017 and May 2018. ctDNA fragments of the target genes (EGFR, KRAS, BRAF, ALK, HER2, and TP53) from the blood samples were enriched by SM-PCR, and DNA libraries were prepared. Finally, a high-throughput sequencing was performed. The EGFR detection of tumor tissue samples was performed using real-time fluorescence PCR based on the amplification refractory mutation system (ARMS) and consistency in the results of EGFR mutation detection in the plasma and tissue was compared. Results:The results of both the methods were consistent (Kappa=0.867, P<0.001). The McNemar's test also indicated that the results are not statistically different (P=0.500). Conclusions: SM-PCR can be used for the detection of plasma EGFR mutations. The target detection sites are more comprehensive and multiple mutations can be detected at the same time. Results of the analysis are more precise and can be absolutely quantified.

Total knee arthroplasty (TKA) is an effective treatment for end-stage knee disease, with the postoperative alignment, component position, soft tissue balance, and prosthesis matching being key factors for the success of TKA. In order to achieve more accurate postoperative alignment and component position, better soft tissue balance and prosthesis matching for longer prosthesis longevity, better postoperative function and higher patient satisfaction, various intelligent accuracy technological aids such as computer assisted navigation (CAN), patient specific instrumentation (PSI), surgical robots, microsensors, customized implants (CI) and personalized 3D preoperative planning have emerged and are given high expectation. In this paper, the authors review the application and research progress of the above technological aids mainly from aspects of alignment, component position, clinical outcomes and cost analysis, so as to provide a reference for the application of related technological aids in TKA.

BACKGROUND: Human immunodeficiency virus (HIV)-negative plasmablastic lymphoma (PBL) is a rare entity of diffuse large B-cell lymphoma (DLBCL). The clinicopathological features of and optimal treatment for HIV-negative PBL remain largely unknown. METHODS: To gain insight into this distinct lymphoma, we summarized the clinicopathologic characteristics of 8 unpublished HIV-negative PBLs and performed a comprehensive review of 394 published cases. RESULTS: Of the 8 unpublished PBLs, the median patient age was 53.0 years. Four patients presented with stage IV disease. All 8 patients showed a plasma cell-like immunophenotype. Of the six patients who received anthracycline-based chemotherapy, including two who received bortezomib, three patients achieved a continuous complete response, two patients died due to disease progression, and one patient was lost to follow-up. The other two patients achieved continuous complete response after receiving chemotherapy combined with radiotherapy and surgery. Of the 402 patients, the majority were male, with a mean age of 58.0 years. EBV infection was detected in 55.7% of the patients. The median survival times of the patients who received CHOP or CHOP-like regimens and intensive regimens were not reached and 23.0 months, respectively, and the intensive regimen did not improve the survival outcome (P=0.981). Multivariate analysis showed that EBER remained the only independent factor affecting overall survival (OS). CONCLUSION HIV-negative PBL is a distinct entity with a predilection for elderly and immunosuppressed individuals. Intensive chemotherapy had no apparent survival benefits over the CHOP regimen in terms of OS; the prognosis of this disease is poor with current chemotherapy methods, and treatment remains a challenge.

Cardiomyocytes can resist ischemia/reperfusion (I/R) injury through ischemic postconditioning (IPoC) which is repetitive ischemia induced during the onset of reperfusion. Myocardial ischemic preconditioning up-regulated protein 2 (MIP2) is a member of the WD-40 family proteins, we previously showed that MIP2 was up-regulated during ischemic preconditioning (IPC). As IPC and IPoC engaged similar molecular mechanisms in cardioprotection, this study aimed to elucidate whether MIP2 was up-regulated during IPoC and contributed to IPoC-mediated protection against I/R injury. The experiment was conducted on two models, an in vivo open chest rat coronary artery occlusion model and an in vitro model with H9c2 myogenic cells. In both models, 3 groups were constituted and randomly designated as the sham, I/R and IPoC/hypoxia postconditioning (HPoC) groups. In the IPoC group, after 45 min of ischemia, hearts were allowed three cycles of reperfusion/ischemia phases (each of 30 s duration) followed by reperfusion. In the HPoC group, after 6 h of hypoxia, H9c2 cells were subjected to three cycles of 10 minute reoxygenation and 10 minute hypoxia followed by reoxygenation. IPoC significantly reduced the infarct size, plasma level of Lactate dehydrogenase and creatine kinase MB in rats. 12 h after the reperfusion, MIP2 mRNA levels in the IPoC group were 10 folds that of the sham group and 1.4 folds that of the I/R group. Increased expression of MIP2 mRNA and attenuation of apoptosis were similarly observed in the HPoC group in the in vitro model. These effects were blunted by transfection with MIP2 siRNA in the H9c2 cells. This study demonstrated that IPoC induced protection was associated with increased expression of MIP2. Both MIP2 overexpression and MIP2 suppression can influence the IPoC induced protection.
Animals ; Blotting, Western ; Cell Hypoxia/genetics/physiology ; Cell Line ; Cell Survival/genetics/physiology ; Flow Cytometry ; Ischemic Preconditioning, Myocardial/*methods ; Male ; Myocytes, Cardiac/*metabolism/*pathology ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Reperfusion Injury/*metabolism/*prevention & control