OBJECTIVETo investigate the relationship between PPAR coactivator 1 (PGC-1), nuclear respiratory factor (NRF), mitochondrial transcription factor A (mtTFA) expressions of vascular smooth muscle cells (VSMC) and development of atherosclerosis in a rabbit model.

METHODSAtherosclerotic model was established by feeding the rabbits with high-fat diet for 4, 8 and 12 weeks (n = 10 each). Another 8 rabbits fed with normal diet served as normal controls. Intima-media ratio, mRNA and protein expressions of PGC-1, NRF, mtTFA and SMemb, a marker for synthetic VSMC, were detected on aorta specimens.

RESULTSWith the blood lipid increased, the intima-media ratio rose from (0.031 +/- 0.010) microm up to (0.814 +/- 0.258) microm during 12 weeks. Increasing SMemb means that synthetic VSMC grew more and more. The expressions of PGC-1 became significant after 4 weeks (P < 0.01), while that of NRF-1 and mtTFA rose significantly after 8 weeks (P < 0.01).

CONCLUSIONSThe PGC-NRF-mtTFA pathway might play a critical role in VSMC proliferation and development of atherosclerosis.

Animals ; Atherosclerosis ; blood ; metabolism ; pathology ; DNA-Binding Proteins ; metabolism ; Disease Models, Animal ; Female ; Lipids ; blood ; Male ; Mitochondrial Proteins ; metabolism ; Muscle, Smooth, Vascular ; metabolism ; Nuclear Respiratory Factor 1 ; metabolism ; Rabbits ; Trans-Activators ; metabolism ; Transcription Factors ; metabolism

BACKGROUND: With the wide application of the proximal closing wedge osteotomy in treatment of hallux valgus deformity, the metatarsal shortening and metatarsal elevation are constantly being mentioned. Scholars even think that the operation is complicated, the technical requirements are high, and the incidence of complications is high, and other osteotomy should be used instead. However, through literature research and clinical experience, it is found that the facts are not completely the case. OBJECTIVE: To perform geometric study of the plane and the physical model of the proximal closing wedge osteotomy so as to reasonably evaluate the effect of this technique on metatarsal shortening and metatarsal elevation. METHODS: The research was divided into four stages, in which the first three stages were plane geometric researches, with weight-bearing frontal image of the patient with hallux valgus as the research materials; the fourth one was solid geometric research, with plaster model of the first metatarsal bone as the research material. In research I (Stage I), the changes in 1/2 inter-metatarsal angle, length of the first metatarsal and distal metatarsal articular angle were learnt through angle adjustment after clipping when the distance from the wedge osteotomy vertex to medial border of the first tarsometatarsal joint was different (B, A, C, D respectively from the near to the distant, with a distance of 10 mm). In research II (Stage II), the changes in 1/2 inter-metatarsal angle, length of the first metatarsal and distal metatarsal articular angle were learnt through angle adjustment after clipping when the distance from the wedge osteotomy vertex to medial and lateral connection of the first tarsometatarsal joint was same, and the distance from the wedge osteotomy vertex to axis of the first metatarsal bone was different. In research III (Stage III), the changes in the above-mentioned research indexes were studied through angle adjustment after clipping when the wedge osteotomy vertex and wedge angle were fixed, but the proximal osteotomy line slope was different. Research IV (Stage IV) was the simulated osteotomy comparison, osteotomy perpendicular to metatarsal backbone or perpendicular to the horizontal plane was conducted on the model through plaster casting, and then osteotomy section was closed; the changes in elevation of the metatarsal head, 1/2 inter-metatarsal angle and length of the first metatarsal bone were measured. RESULTS AND CONCLUSION: (1) Results of plane geometry: The closer the wedge osteotomy vertex was to the proximal end, the larger the correction gained was and the smaller the changes in the distal metatarsal articular angle were, but the shortening was more obvious. Moreover, the closer the wedge osteotomy vertex was to the axis of the metatarsal bone, the smaller the shortening was; if only the proximal osteotomy line was different, the indices were the same. (2) Results of stereoscopic geometry: In the case of the same osteotomy angle, the cuneiform bone of the same angle was intercepted, and the degree of correction was obviously greater than that of the plane geometry. This should be related to the thickness of the osteotomy saw and the loss of bone mass caused by the osteotomy. The metatarsal bone was significantly higher after correction when perpendicular to the metatarsal stem compared with that perpendicular to the horizontal surface. In 1/2 inter-metatarsal angle, the difference between the two was only 1°. In metatarsal shortening, it was smaller when perpendicular to the metatarsal stem compared with that perpendicular to the horizontal surface. (3) In conclusion, a. the optimal wedge osteotomy vertex is not at the medial border of the joint but is 1 cm from the medial border of the joint; then, it will approach the axis of the metatarsal. In this way, good correction, steady fixation and further reduction of metatarsal shortening can be achieved. b. The optimal direction of the oscillating saw blade is perpendicular to horizontal plane; however, an approximately perpendicular position to the horizontal plane is acceptable. It does not affect the effect of orthopedics, and further reduces the metatarsal shortening. When the bony closure is closed, the distal end of the metatarsal bone is taken to avoid the elevation of the metatarsal bone with a 2 mm step with the proximal end. c. The angle selection of proximal osteotomy surface can be placed on the metatarsal stem according to the internal fixation condition, and it can also form an acute angle with the metatarsal stem, so that the effect of osteotomy will not be changed.

OBJECTIVETo explore the possibility of using melanoma antigen (MAGE)-1 and MAGE-3 gene encoding proteins as an index of potential target for immunotherapy in intrahepatic cholangiocarcinoma (IHCC) patients.

METHODSThe expressions of MAGE-1 and MAGE-3 genes in tumor tissues and tumor adjacent non-IHCC liver tissues were examined by RT-PCR method. The relationship between positive expression rates of MAGE-1 and MAGE-3 genes and clinical data including sex, age, tumor diameters, tumor envelope, tumor nodules number, and hepatitis B virus surface antigen were determined.

RESULTSThe positive expression rates of MAGE-1 (35%) and MAGE-3 genes (45%) were significantly higher in the tumor tissues than in tumor adjacent tissues (0) (P<0.01). The positive expression rates of MAGE-1 and MAGE-3 genes had no relationship with the clinical data (P >0.05), except the morphology of tumor (P <0.05).

CONCLUSIONThe high expression rates of MAGE-1 and MAGE-3 genes in IHCC suggests the MAGE-1 and MAGE-3 gene may be a target for immunotherapy in IHCC patients.

Adult ; Aged ; Antigens, Neoplasm ; genetics ; Bile Duct Neoplasms ; genetics ; Bile Ducts, Intrahepatic ; pathology ; Cholangiocarcinoma ; genetics ; Female ; Humans ; In Vitro Techniques ; Liver Neoplasms ; genetics ; Male ; Melanoma-Specific Antigens ; Middle Aged ; Neoplasm Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo study the mechanism of action of Tougu Xiaotong Capsule (透骨消痛胶囊, TGXTC) ex vivo in suppressing chondrocyte (CD) apoptosis induced by sodium nitroprussiate (SNP).

METHODSThirty New Zealand rabbits, 2 months old, were randomized by lottery into five groups, six in each: the blank group treated with saline, the positive control group treated with Zhuanggu Guanjie Pill (壮骨关节丸, 70 mg/kg), and the three experimental groups, EGA, EGB, and EGC, treated with low dose (35 mg/kg), moderate dose (70 mg/kg), and high dose (140 mg/kg) of TGXTC, respectively. All treatments were administered via gastrogavage twice a day for 3 days. Arterial blood was collected from the abdominal aorta and drug or drug metabolites-containing serum was prepared. CDs obtained from knee joints of 16 four-week-old New Zealand rabbits were cultured to the third passage and confirmed by toluidine blue staining. SNP of various final concentrations (0, 0.5, 1.0, and 2.0 mmol/L) was used to induce CD apoptosis, and the dosage-effect relationship of SNP in inducing CD apoptosis was determined. Serum samples from the blank, control, and three dosages of TGXTC-treated rabbits were tested in the CD culture in the presence of SNP. Cell apoptosis was determined by Hoechst 33342 staining, viability of CDs was quantified by MTT, CD apoptosis rate was determined by annexin V-FITC/PI staining, levels of p53 and Bcl-2 mRNA expression in CDs were determined with RT-PCR, and contents of caspase-3 and caspase-9 proteins were determined by colorimetry.

RESULTSCD apoptosis was induced by SNP at all concentrations tested and in a dose-dependent manner. The SNP concentration of 1 mmol/L and treatment duration of 24 h appeared to be optimal and were selected for the study. Serum samples from the positive control rabbits and from the two higher doses of TGXTC-treated rabbits showed reduction of SNP-induced CD apoptosis, decrease in p53 mRNA expression, inhibition of catalytic activities of caspase-3 and caspase-9, and increase in Bcl-2 mRNA expression when compared with the serum from the blank group (P<0.05).

CONCLUSIONTGXTC-containing sera antagonized SNP-induced CD apoptosis and the molecular basis for the action was associated with up-regulation of Bcl-2, down-regulation of p53 expression, and inhibition of caspase-3 and caspase-9 catalytic activities.

Animals ; Apoptosis ; drug effects ; Biocatalysis ; drug effects ; Capsules ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Survival ; drug effects ; Cells, Cultured ; Chondrocytes ; drug effects ; enzymology ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation ; drug effects ; Male ; Models, Biological ; Nitroprusside ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Rabbits ; Reproducibility of Results ; Serum ; chemistry ; Tumor Suppressor Protein p53 ; genetics ; metabolism

OBJECTIVETo analyze expression characteristics of human skin epidermal stem cell at different developmental stages, and to explore its biological significance.

METHODSHealth skin samples from 28-32 w fetuses (F group), 4-12 y children (C group), and 35-55 y adult (A group) were harvested, with 10 cases in each group. Epidermis were separated using trypsin digestion and EDTA, and human epidermal stem cells were isolated and purified with type IV collagen attachment method. The monoclonal antibody of integrin beta1 and keratin 19 were used for detection and identification of epidermal stem cells by immunohistochemical staining. Total RNA was extracted from above cells by Trizol one-step method, and were detected by formaldehyde denaturing agarose gel electrophoresis. Probes were prepared and hybridized into cDNA microarray for scanning fluorescent signals and analysis of images, with two-fold differential expression value for screening. Significantly up/down-regulated genes were selected for verification by real time RT-PCR.

RESULTSBy comparing expression profile between A and C groups, a total of 1808 genes with differential expression were detected, including 1089 up-regulated genes and 719 down-regulated genes, and they were classified into 128 categories. Among them, 1462 genes were known (found in GeneBank), 346 genes were unknown. A total of 4534 genes with differential expression were detected between C and F groups, in which 1783 genes were up-regulated and 2751 genes were down-regulated, and they were classified into 216 categories. Among them, 3577 genes were known (found in GeneBank), and 957 genes were unknown. There were 1104 genes with differential expression consistently detected in F, C and A groups, which were classified into 32 categories according to gene function. Among them, 94 genes were consistently up-regulated and 75 genes consistently down-regulated. Test results of real time RT-PCR were in accordance with above-mentioned results.

CONCLUSIONSGene expression profiles of epidermal stem cells cultured in vitro, harvested from fetuses, children, and adult, exhibit obvious difference. This may be closely related to different stages of proliferation and differentiation of human epidermal stem cell and self-repair ability of wound at different developmental stages.

Adult ; Cell Differentiation ; Child ; Child, Preschool ; Epidermis ; cytology ; growth & development ; Epithelial Cells ; cytology ; Fetus ; cytology ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; Humans ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; methods ; Stem Cells ; cytology ; Transcriptome

OBJECTIVETo observe the precise time of the recurrence after resection of hepatocellular carcinoma (HCC) and to further explore the risk factors associated with postoperative recurrence.

METHODSTotally 94 patients who had undergone resection of HCC were divided into three groups based on the time of recurrence, which was indicated by the digital subtraction angiography (DSA) examination: recurrence between 1 to 6 months, recurrence between 7 to 12 months, and tumor-free after 12 months. Patients with intra-hepatic recurrence were treated with transcatheter arterial chemoembolization and confirmed by CT scans after embolization, contrast-enhanced ultrasound, or magnetic resonance imaging.

RESULTSThe recurrence rates of 6 months and 1 year were 30.9% and 36.2%, respectively. No statistically significant difference between 6-month and 1-year recurrence rates was observed. Nine (26.5%) patients with recurrence and five (8.3%) patients free of tumor had previously presented as multifocal HCC, which showed a statistical significance (P = 0.032). The diagnostic accuracy of DSA was 87.2%, which was eventually confirmed by the other investigations.

CONCLUSIONSMost recurrences occure within the first six months postoperatively and multifocal carcinogenesis is one of the risk factors associated with early recurrence after liver resection for advanced HCC. DSA is an important surveillance for early detection of intra-hepatic recurrence after surgery; meanwhile, it also provides information for early management to control the disease progression and for future active therapies.

Adult ; Aged ; Carcinoma, Hepatocellular ; diagnostic imaging ; pathology ; surgery ; Female ; Hepatectomy ; Humans ; Liver Neoplasms ; diagnostic imaging ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Postoperative Period ; Tomography, X-Ray Computed

OBJECTIVETo establish a three-dimentional liver function evaluation system using 99mTc-diethyl iminodiacetic acid (99mTc-EHIDA) scintigraphy based on single photon emission computed tomography (SPECT).

METHODSTotally 16 patients with liver lesions were divided into cirrhosis group and non-cirrhosis group. SPECT was performed 2 days before operation and 5 days after operation. Serum liver functions were examined on the same day of scintigraphy. SPECT images of areas of interest of heart and liver were aquired. Time of the peak of EHIDA density in liver (Tpeak), five-minutes heart liver index (HLI5), blood clearance index (HH15), receptor index (LHL15), and the predictive values were calculated.

RESULTSTpeak was not significantly different between two groups, while HLI5, HH15, and LHL15 were significantly different (P = 0.033, P = 0.001, and P = 0.005). HLI, and LHL15 were significantly correlated with preoperative total protein and prealbumin levels (P = 0.003, P = 0.015, P = 0.022, P = 0.038) and post-operative prealbumin (P = 0.037, P = 0.042). The predictive values of HLI5 and LHL15 correlated well with postoperative HLI5 and LHL15 (r = 0.675, P = 0.016; r = 0.629, P = 0.028).

CONCLUSIONThe three-dimentional liver function evaluation system using 99mTc-EHIDA based on liver SPECT may facilitate the further studies of risks of liver surgery.

Adult ; Aged ; Animals ; Female ; Humans ; Liver Diseases ; diagnosis ; diagnostic imaging ; physiopathology ; Liver Function Tests ; Male ; Middle Aged ; Postoperative Period ; Preoperative Period ; Radiopharmaceuticals ; administration & dosage ; Technetium Tc 99m Diethyl-iminodiacetic Acid ; administration & dosage ; Tomography, Emission-Computed, Single-Photon

OBJECTIVETo summarize the surgical experiences, risks, complications, and managements for hepatic masses in difficult sites.

METHODSTotally 47 patients were divided into three groups based on the liver tumor sites: primary porta hepatis group, secondary porta hepatis group, and caudate lobe group. All patients underwent different portion of hepatectomy.

RESULTSThe surgery duration was (289.6 +/- 62.2) ml-nutes, intra-operative blood loss was (602.3 +/- 256.4) ml, and intra-operative blood transfusion was (524.0 +/- 325.9) ml. Incidence of surgical complications in each group was 61.5%, 26.9%, and 25%, respectively. Serious complications observed were biliary leakage (27.7%), bleeding (6.4%), and post-operative liver failure (2.1%). Three perioperative deaths were reported: two patients died of bleeding, and one patient died from liver failure.

CONCLUSIONSDissection of the liver and exposure of major blood vessels and biliary ducts are of critical importance in the surgeries for hepatic masses in difficult sites, and post-operative complications may be remarkably reduced through delicate manipulations of the small intra-hepatic vessels and biliary ducts during resection. A thorough pre-operative evaluation plays a key role in predicting the feasibility and risks of the surgery. Damage to the major blood vessels adjacent to the tumor, in addition to bleeding, may result in in-flow or outflow obstruction and cause necrosis of the corresponding hepatic lobe. Compared with damage to the primary portal area, vascular damage to the secondary porta is generally associated with higher fatality.

Adult ; Aged ; Blood Loss, Surgical ; Female ; Hepatectomy ; adverse effects ; Humans ; Liver Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Postoperative Complications ; Preoperative Care

Adult ; Female ; Humans ; Liver Neoplasms ; diagnosis ; surgery ; Neoplasms, Germ Cell and Embryonal ; diagnosis ; surgery ; Sarcoma ; diagnosis ; surgery ; Tomography, X-Ray Computed

OBJECTIVETo study the toxicity features of high glucose on the endothelial cell cycle and the influence of Dan Gua-Fang, a Chinese herbal compound prescription, on the reproductive cycle of vascular endothelial cells cultivated under a high glucose condition; to reveal the partial mechanisms of Dan Gua-Fang in the prevention and treatment of endothelial injury caused by hyperglycemia in diabetes mellitus (DM); and offer a reference for dealing with the vascular complications of DM patients with long-term high blood glucose.

METHODSBased on the previous 3-(4,5)-dimethylthiahiazo (z-y1)-3-5-diphenytetrazoliumromide (MTT) experiment, under different medium concentrations of glucose and Dangua liquor, the endothelial cells of vein-304 (ECV-304) were divided into 6 groups as follows: standard culture group (Group A, 5.56 mmol/L glucose); 1/300 herb-standard group (Group B); high glucose culture group (Group C, 16.67 mmol/L glucose); 1/150 herb-high glucose group (Group D); 1/300 herb-high glucose group (Group E); and 1/600 herb-high glucose group (Group F). The cell cycle was assayed using flow cytometry after cells were cultivated for 36, 72 and 108 h, respectively.

RESULTS(1) The percentage of cells in the G0/G1 phase was significantly increased in Group C compared with that in Group A (P<0.05), while the percentage of S-phase (S%) cells in Group C was significantly reduced compared with Group A (P<0.05); the latter difference was dynamically related to the length of growing time of the endothelial cells in a high glucose environment. (2) The S% cells in Group A was decreased by 30.25% (from 40.23% to 28.06%) from 36 h to 72 h, and 12.33% (from 28.06% to 24.60%) from 72 h to 108 h; while in Group C, the corresponding decreases were 23.05% and 21.87%, respectively. The difference of S% cells between the two groups reached statistical significance at 108 h (P<0.05). (3) The percentage difference of cells in the G2/M phase between Group C and Group A was statistically significant at 72 h (P<0.01). (4) 1/300 Dan Gua-Fang completely reversed the harmful effect caused by 16.67 mmol/L high glucose on the cell cycle; moreover it did not disturb the cell cycle when the cell was cultivated in a glucose concentration of 5.56 mmol/L.

CONCLUSIONSHigh glucose produces an independent impact on the cell cycle. Persistent blocking of the cell cycle and its arrest at the G0/G1 phase are toxic effects of high glucose on the endothelial cell cycle. The corresponding variation of the arrest appears in the S phase. 1/300 Dan Gua-Fang completely eliminates the blockage of high glucose on the endothelial cell cycle.

Cell Cycle ; drug effects ; physiology ; Cells, Cultured ; Culture Media ; pharmacology ; Cytoprotection ; drug effects ; Dose-Response Relationship, Drug ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Cells ; drug effects ; physiology ; Flow Cytometry ; Glucose ; adverse effects ; Humans