OBJECTIVETo explore the relationship of CD44 expression or glycosylation and hepatocellular carcinoma (HCC) metastasis.

METHODSIHC, Quantum dots detection, RT-PCR, Western blot, Cellular immune fluorescence and MS-PCR were used to identify CD44 expression in HCC samples and a series of human HCC cell lines with different metastatic potentials. Lectin array was used to reveal the relationship of CD44v6 glycosylation and human HCC metastasis.

RESULTSImmunohistochemistry analysis showed that CD44v6 was mainly distributed on the cell membrane, while CD44S immunoreactivity was prominently in the cytoplasm, CD44v3 and CD44v4/5 were in cytoplasm on membrane. Among CD44S and those CD44 variants, only the expression of CD44v6 was higher in metastasis HCC samples as compared to that in non-metastasis group (x²=8.828, P less than 0.05). This result was also re-confirmed by the result of Quantum dots (t = 2.392, P < 0.05) and serum detection (t = 2.56, P < 0.05). We found completely methylation of CD44v6 gene in Hep3B and incomplete methylation in MHCC97H and MHCC97L cell lines with metastatic potentials. The lectin affinity assay indicated that lectin MAL, SNA and WGA showed more affinity to MHCC97H and MHCC97Lcell lines than that of the non-metastatic Hep3B cell lines.

CONCLUSIONSCD44v6 over-expression presents a positive correlation with HCC metastatic potential, which may be associated with DNA methylation level in promoter sequence. The increasing sialic acid modified glycan of CD44v6 might be related to HCC metastatic ability.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; DNA Methylation ; Glycosylation ; Humans ; Hyaluronan Receptors ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; pathology ; Promoter Regions, Genetic

AIM: To observe of optical coherence tomography angiography (OCTA) and indocyanine green angiography (ICGA) image feature in polypoidal choroidal vasculopathy (PCV).METHODS: Selected 21 patients 21 eyes with PCV in our hospital from January 2016 to December 2016.All the eyes were examined by ICGA, and was examined by OCTA after ICGA examination 1h.We observed the characteristics of OCTA and ICGA images.RESULTS:ICGA examination showed that there were 8 cases of choroidal abnormal branch vascular network (BVN), polypoid lesions 10 eyes, BVN with polypoid lesions 2 eyes, no abnormal performance 1 eyes.OCTA examination showed 8 eyes of BVN, and the location, range and shape of BVN were similar to ICGA in OCTA examination.ICGA examination showed 10 cases of polypoid lesions.OCTA showed strong signal highlights.ICGA examination showed 2 cases of BVN complicated with polypoid lesions, and OCTA examination showed strong signal highlights of BVN and corresponding parts.ICGA examination showed no abnormal performance in 1 eyes, and no abnormal findings in OCTA examination.CONCLUSION: OCTA and ICGA are similar in the location and morphology of PCV lesions, and OCTA may play a role in the diagnosis of PCV restricted ICGA.

Objective To identify genes differentially expressed in the window of implantation and explore the molecular basis of the development of endometrial receptivity.Methods A subtracted cDNA library of the window of implantation was constructed by suppression subtractive hybridization(SSH) method.The screened clones of the subtracted library were sequenced and GenBank homology search was performed.The differential expression of ribosomal protein(RP)L7,RPL7 pseudogene(RPL7p),RPL19 and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta polypeptide(YWHAZ) were further confirmed by RT-PCR.Results After sequencing and GenBank homology search of 50 clones, 35 differentially expressed genes were detected in the window of implantation,of which 23 were known genes,and 12 were unknown genes.Some of the known genes have been proved to be associated with implantation,while others were firstly screened out by us.The results of RT-PCR confirmed that RPL7, RPL7p,RPL19 and YWHAZ were highly expressed in the window of implantation,0.75?0.21,1.72? 0.30,1.23?0.31,and 1.28?0.08,respectively.Conclusions SSH is a useful technique to detect differential expression genes and an effective method to clone novel genes.It provides a new method to investigate the molecular basis of the development of endometrial receptivity.

OBJECTIVETo test expression level and glycosylation level of OPN in HCC cell lines with different metastatic potential and HCC tissues, and investigate the correlation between the glycosylation change and the liver cancer transporting as well as its significance.

METHODSThe level of OPN expression in liver cancer tissue(6 cases of non-metastasis and 7 cases of metastasis)as well as HCC cell lines with different metastatic potential (L02, Hep3B, MHCC97L, MHCC97H, HCCLM3, HCCLM6)was identified by immunohistochemistry and Western Blot, and then OPN was purified from HCC tissues by immunoprecipitation, followed by glycosylation detection of OPN from non-metastatic and metastatic HCC tissues by multiple lectin blot. Data were analyzed by t-test and variance analysis.

RESULTSDifferent levels of OPN expression were observed in HCC cell lines with different metastatic potential (F = 5.04, P = 0.008). Additionally, OPN expression level in HCC tissues with metastasis was higher than that in non-metastasis group (t = 2.447, P < 0.05). Relative optical density value was 0.69 ± 0.21 and 0.45 ± 0.14 respectively. OPN in liver cancer tissue was successfully purified using immunoprecipitation. Followed lectin blotting result showed that OPN protein in metastasis group showed lower affinity to MAL, PHAE, DSA, ConA as compared with that in non-metastasis group (P < 0.05).

CONCLUSIONSThe expression of OPN was positively correlated with the enhanced metastasis potential of HCC. OPN from metastasis HCC tissues presented lower level of some specific glycan structures such as a2, 3- sialic acid, bisecting GlcNAc, biantennary, muti-antennary and high mannose type N-glycan structure. This study not only indicates the role of OPN in HCC metastasis for the first time, but also provide experimental support for the mechanism of the function of OPN in the transportation of liver cancer cells as well as offer potential target for clinical treatment.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Glycosylation ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; Osteopontin ; metabolism

OBJECTIVETo develop an early and accurate detection method for hepatocellular carcinoma (HCC) based on detection of tumor-associated serum markers using a multiplex quantitative antibody array.

METHODSThe double-antibody sandwich principle was used to establish an antibody array composed of eight cancer-related serum markers, including alpha-fetoprotein (AFP), hepatocyte growth factor (HGF), insulin-like growth factor (IGF), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), transforming growth factor-beta 1 (TGF-b1), and vascular endothelial growth factor (VEGF). Serum samples from 160 cases of clinically diagnosed HCC and from 58 cases of liver cirrhosis (LC; controls) were obtained to test the array. Sixty percent of the samples were randomly selected for use as the training set (HCC, n = 96; LC, n = 36), and the remaining 40% was used as the test set (HCC, n = 64; LC, n = 22). The SPSS statistical software was used to perform logistic regression analysis and to create a diagnostic model.

RESULTSWhen used with the training set, the model had sensitivity of 93.3%, specificity of 83.3%, and accuracy of 90.9%. When used with the test set, the model had sensitivity of 89.0%, specificity of 77.3%, and accuracy of 86.0%. The traditional serum AFP value (cut-off value of 20 ng/mL) showed 70.0% diagnostic sensitivity, 59.0% specificity, and 64.0% accuracy.

CONCLUSIONThe newly developed multiplex quantitative antibody detection system has high sensitivity and specificity. The diagnostic model with AFP and seven other cancer-related factors was superior to the traditional AFP only approach for early diagnosis of liver cancer, indicating its potential clinical value.

Adult ; Aged ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; diagnosis ; Early Diagnosis ; Female ; Humans ; Liver Neoplasms ; diagnosis ; Male ; Microchip Analytical Procedures ; Middle Aged ; Sensitivity and Specificity ; Young Adult ; alpha-Fetoproteins ; immunology

To explore the biological function and possible underlying mechanism of stathmin gene during hepatocarcinogenesis. Three pairs of chemically synthesized small interfering RNA (siRNA) targeting on stathmin were transfected into HCCLM3 by LipofectamineTM 2000. After confirming the interfering effects of stathmin siRNAs through reverse transcription PCR and Western blotting, the HCCLM3 cells proliferation and apoptosis were detected by cell count kit-8 (CCK-8) and flow cytometry analysis, and the expressions of tumor-related genes (c-myc, c-fos, p53, etc) were observed by real-time PCR. Stathmin expression was effectively inhibited up to 90% by stathmin silencing in HCCLM3 cells (P is less than to 0.05) . By using CCK8 assay, it was shown that HCCLM3 cells proliferation were obviously depressed by 13.04%+/-0.10%, 28.10%+/-0.41% and 37.36%+/-2.15% at the time point of 24 h, 48 h and 72 h with the comparison to Mock group (F = 4.21, P is less than to 0.05). The results of flow cytometry demonstrated that the percentage of apoptotic cells was increased to 25.11%+/-1.62% in RNAi group, compared with 9.20 %+/-0.64 % in Mock group (F = 44.67, P is less than to 0.01). The results of real-time PCR showed that oncogenes c-myc and c-fos expressions were repressed, proliferation-associated gene ki-67 was down-regulated, and apoptosis-promoting gene caspase-3, bax and p53 were induced (P is less than to 0.05). Stathmin may promote cell proliferation, inhibit cell apoptosis and induce malignant transformation of hepatocytes by regulating some tumor-related genes expressions.
Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; Stathmin

objective To understand the baseline data of mortality from malignant tumor from 2006 to 2009 around Hongyanhe Nuclear Power Plant in Wafangdian City,Liaoning Province,so as to provide scientific basis for evaluating the impact of normal operation of nuclear power plant on the health of the residents nearby.Methods Thirty small towns near Hongyanhe Nuclear Power Plant were divided into 5 investigated areas according to the distances away therefrom(0.,10.,20-,30-,and 40-km).The data about from malignant tumor were obtained from the Center for Disease Control and Prevention of Wafangdian.The mortality distribution of difierent malignant tumors was analyzed,including the radiosensitive malignant tumors,especially leukemia,breast cancer,and thyroid gland cancer in different area,gender,and age groups.Results The mortality from malignant tumor was 151.97/105,and the standardized mortality rate(SMR)was 97.76/105.The mortality from malignant tumor among the males was 188.28/105(with the SMR of 116.76/105),and that among the females was 113.47/105(with the SMR of 75.89/105).with a sex ratio of 1.71.The first five cancers in the rank of death causes were lung,liver,stomach,colorectal,and esophageal cancers with mortality of 46.19/105,23.51/105,20.30/105,8.06/105 and 5.45/105,respectively.The mortality from mal.ignant tumor in the areas around the nuclear power plant from the near to the distant were 99.85/105, 137.40/105,138.73/105,156.30/105,and 154.16/105,respectively.The mortality from radiosensitive malignant tumors,leukemia,breast cancer,and thyroid gland cancer were 4.57/105,4.06/105,and 0.26/105,respectively.Conclusions Lung cancer and digestive tract malignant tumors are the main causes of death from malignant tumors in Wafangdian area before the nuclear power plant began to operate.There are no significant differences in the mortality distribution of malignant tumors among different areas,genders,and age groups.There are not significant differences in the mortality distribution of leukemia and breast cancer among different areas and age groups.

ObjectiveTo analyze the causes of occupational acute 1,2-dichloroethane (1,2-DCE) poisoning accident during the use of polyurethane grouting materials for waterproof plugging operation in the construction industry. Methods By combining the clinical symptoms of the patient, worksite survey of occupational health and workplace occupational hazards monitoring method, the cause of an occupational acute 1,2-DCE poisoning accident was investigated at a construction site during the use of polyurethane grouting material for waterproofing and plugging operations. Results The patient was engaged in waterproof grouting work using polyurethane grouting material. The main volatile organic components in the raw materials were 1,2-DCE, with traces of dichloromethane, methyl acetate and others. The result of post-incident on-site investigation showed that the short-term exposure concentration of 1,2-DCE in the workplace air was 578.70 mg/m³, which was more than 30 times higher than the national occupational health standard limit. The mass concentration of 1,2-DCE in the patient's blood was 230 μg/L. Combined with the patient's occupational hazard exposure history, clinical manifestations, worksite survey of occupational health, and laboratory test results, according to GBZ 39-2016 Diagnosis of Occupational Acute 1,2-Dichloroethane Poisoning, this incident was diagnosed as a severe occupational acute 1,2-DCE poisoning event caused by the use of inferior polyurethane grouting material. Conclusion The excessive concentration of 1,2-DCE in the workplace air is the main cause of this poisoning accident. Construction sites with confined space operations should improve various occupational health management systems, occupational health engineering protective facilities, and personal protective equipment must be provided for workers.

OBJECTIVETo discuss efficacy of the surgical treatment strategy of double disruption of the superior shoulder suspensor complex (SSSC).

METHODSThe data of 15 patients with double disruption of the SSSC were retrospectively analyzed from January 2008 to December 2009. There were 11 males and 4 females, with an average age of 45.1 years (ranged, 19 to 60). Of them, 8 patients were treated with surgery and 7 patients with conservative treatment. The short-term effectiveness was evaluated according to Constant-Murley scoring system.

RESULTSAll patients were followed up for 7 to 24 months with an average of 14.4 months. All fractures healed with a mean time of 12.3 weeks (ranged, 9 to 12). At final follow-up, the Constant-Murley scores with patients of surgical treatment was (92.37 +/- 1.99), and patients of conservative treatment was (55.52 +/- 1.29).

CONCLUSIONSurgical treatment can restore the integrality of SSSC, in favour of stability between upper limb and trunk.

Adult ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Shoulder Fractures ; physiopathology ; surgery ; Shoulder Joint ; injuries ; physiopathology ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo observe the therapeutic efficacy of dietary boron supplement on retinoic acid-induced osteoporosis in rats, so as to provide experimental evidence for clinical management of osteoporosis with boron.

METHODSThirty-two SD rats were randomized into normal control group (8 rats) and osteoporotic group (24 rats), and osteoporosis was induced in rats of the latter group by intragastric retinoic acid administration at the daily dose of 80 mg/kg for 15 consecutive days. The osteoporotic rats were subdivided into control group (8 rats) without treatment, boron treatment group (8 rats) and estradiol treatment group (8 rats). After 30 days of treatment, the serum contents of Ca, P, boron and the activities of alkaline phosphatase (AKP) and tartrate-resistant acid phosphatase (TRAP) in the rats were assayed, the bone mineral density (BMD) of the whole body, lumbar vertebrae and tibia were determined, and the morphological changes of the femurs were observed.

RESULTSThe serum contents of Ca and P in the rats of the 4 groups differed scarcely, but the content of boron in boron treatment group was markedly higher than that in the other three groups. In the osteoporotic control group, the activities of serum AKP and TRAP, the masses of spongy bone and cortical bone of the femurs, and the quantity of the osteoclasts were increased, with the BMD of the lumbar vertebrae and tibia decreased, suggesting osteoporotic conditions. The mean trabecular plate density and thickness, trabecular bone volume and cortical bone volume of the femurs in the osteoporotic rats treated with boron or estradiol were significantly increased, but the active osteoclast quantity in the spongy bone and serum TRAP activities were obviously decreased, and the bone quality was comparable with that of the normal group. In addition, the serum AKP activity and the active osteoblast quantity in the spongy bone were obviously increased in boron treatment group.

CONCLUSIONThe dietary boron supplement can increase the serum content of boron of osteoporotic rats to stimulate bone formation and inhibit bone resorption, producing therefore obvious therapeutical effect against osteoporosis.

Acid Phosphatase ; blood ; Alkaline Phosphatase ; blood ; Animals ; Bone Density ; drug effects ; Boron ; administration & dosage ; therapeutic use ; Dietary Supplements ; Female ; Femur ; drug effects ; growth & development ; metabolism ; Isoenzymes ; blood ; Osteoporosis ; blood ; chemically induced ; drug therapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tartrate-Resistant Acid Phosphatase ; Time Factors ; Tretinoin