Cancer metastasis is multi-step process that requires cancer cells to jump through many hoops.The low-molecule-weight beparin (LMWH) has been reported effective for the treatment of metastasis to some extent, the mechanisms are proposed as anticoagulation, inhibition of heparanase, selectins, adhesion, angiogenesis mediated by the tumor cells, and the effects on cell cycle and apoptosis.

Objective To compare the results of anti-cardiolipin (ACA)measuring by two commercial available kits.Methods ACA in total of 66 serum samples were both determined by kits from Euroimmun and YHLO simultaneously,then the re-sults were analyzed comparatively and correlatively.The Euroimmun kit was applied to determination the level of ACA-IgA/G/M,and the YHLO kit determined ACA-IgG/M and anti-β2-glycoprotein I antibody (β2GPI IgG).Results The positive rate by Euroimmun kit was 37.88% (25/66),while 31.82% (21/66)was positive (one positive among ACA-IgG,IgM andβ2GPI IgG)when determined by YHLO kit,and there was no significant difference between the two kits.The accordance rate of the two kits was 87.88% (58/66).The ACA-IgA/G/M value by Euroimmun kit and the summation of ACA-IgG, IgM andβ2GPI IgG by YHLO kit showed well linear correlation (r 2 =0.892,P <0.01).Conclusion Results from the two kits were consistent and correlated well,and they are suitable for the clinical application;these two kits have their own char-acteristics,which could be used by individual or combination accordingly.

BACKGROUND:Procolagen type 1 N-terminal propeptide (P1NP) and β-colagen special sequence(β-CrossLaps) are two bone metabolic markers that are closely related to osteoporosis. Combined detection of bone metabolic markers and bone mineral density is of clinical significance for the diagnosis of osteoporosis. Bone metabolic markers are ideal indicators to predict fractures, which can compensate for the lack of bone density test. OBJECTIVE:To introduce the application of bone metabolic markers in the monitoring of drug efficacy on the treatment of osteoporosis as wel as in the prediction of fracture risks in recent 20 years and to explore the clinical values of P1NP and β-CrossLaps to assess the therapeutic efficacy on osteoporosis and risks for osteoporotic fractures. METHODS:A computer-based search of CNKI and SCI databases were performed for relevant articles published from 2000 to 2014 using the keywords of “serum bone metabolic markers; osteoporosis; bone mineral density” in Chinese and English, respectively. Finaly, 44 articles meeting the inclusive criteria were reviewed. RESULTS AND CONCLUSION:This paper analyzes the source and detection mechanisms of P1NPand β-CrossLaps and then compares their advantages in the therapeutic effect assessment of osteoporosis. Serum bone metabolic markers cannot only reflect the dynamic changes of bone metabolism, but also have earlier changes than the bone mineral density. Both P1NPand β-CrossLaps are very important for assessing the early diagnosis of osteoporosis as wel as anti-osteoporosis drug efficacy.

BACKGROUND: Ankle fracture combined with deltoid ligament rupture and distal tibiofibular syndesmosis injury occurs occasionally. Its treatment with distal tibiofibular syndesmosis screw fixation or deltoid ligament repair remains controversial. The former appears with poor reduction, broken nails, secondary surgery and other problems.OBJECTIVE: To observe the clinical effectiveness of suture anchor repair for ankle fracture combined with deltoid ligament injury.METHODS: Twelve patients with ankle fracture combined with deltoid ligament injury were selected from the First Affiliated Hospital of Guangzhou University of Chinese Medicine between January 2013 and December 2015. All patients were treated with open reduction, internal fixation, and anchor repair, but without distal tibiofibular syndesmosis screw fixation. The curative efficacy and joint stability were observed.RESULTS AND CONCLUSION: (1) All patients were followed-up for more than 12 months. (2) The modified Baird-Jackson scoring system showed excellent in nine cases, good in two cases, average in one case, poor in none case, and the excellent and good rate was 92%. (3) At 1 year after internal fixation, the X-ray examination showed the malleolus gap and talus slope angle of the affect side were (3.47±0.12) mm and (0.45±0.18)°, and the malleolus gap and talus slope angle of the healthy side were (3.44±0.05) mm and (0.43±0.14)°, and there was no significant difference between two sides (P > 0.05). (4) These results indicate that the suture anchor can repair the anatomy and biomechanics of deltoid ligament with stable ankle joint, and secondary surgery is unnecessary.

Objective To investigate manifestations of connective tissue nevi (CTN) by dermoscopy and reflectance confocal miscroscopy (RCM),and to analyze its pathological features.Methods According to the course of the disease,skin lesions of a patient with CTN were classified into 2 categories:skin lesions of less than 1-year duration and those of more than 1-year duration,which were observed by dermoscopy and RCM,and analyzed by the combination with histopathological examination.Results Dermoscopy imaging of the CTN lesions of more than 1-year duration showed grey-white maculae with clear boundaries in the center of lesions.There were white network structures,brown globules,red spots and punctiform or comma-shaped blood vessels in the gray-white macula area,and vellus hairs and dermal papillary rings could be seen sometimes.The white macula was evenly surrounded by brown to black pigments,and the boundary was clear.For the lesions of less than 1-year duration,gray-white maculae with clear boundaries were also found in the center of the lesions.Besides,there were a few scattered brown to black pigments,vellus hairs and dermal papillary rings in the gray-white macula area.RCM imaging of the lesions of less than 1-year duration or early skin lesions showed dermal papillae of different sizes and shapes crowded with each other and formed a honeycomb or cobblestone-like structure.The density and brightness of the dermal papilla ring obviously increased,and the refraction of the dermal collagen fiber bundles was also enhanced.Some of the dermal collagen fiber bundles were attached to the dermal papilla or intertwined and arranged in a whirlpool pattern.For the lesions of more than 1-year duration,the refraction of the tissue structure was weakened when the depth in Z-axis direction reached 2 to 3 millimetres due to the thickened epidermis,while the dermal collagen fiber bundles were not obvious and only a few lymphocytes could be seen.Conclusions CTN has particular characteristics in dermoscopy,and there is a definite correspondence among dermoscopy features,histopathological findings and RCM features.RCM is helpful for the diagnosis of lesions of less than 1-year duration.

Abstract： Objective　To elucidate the antigenic antibody reaction of recombinant expression of non-structural protein 1 (NS1) of Japanese encephalitis (JE) virus with various mosquito-borne flaviviruses, including JE virus, and the antigenic antibody reaction of serum samples of patients infected with JE virus in acute stage. Methods　In this study, Escherichia coli prokaryotic expression vector (pET) system was used to recombinant express Japanese encephalitis virus NS1 gene. Western Blot assay was performed to detect the antibody responses of the recombinantly expressed protein against a variety of mosquito-transmitted flaviviruses, including JE virus, as well as antigen-antibody reactions of serum from patients with acute JE virus infection. Results　The NS1 gene expression product of JE virus (P3 strain) was in the form of an inclusion body, and the denatured and renatured expression product was displayed as a single band in the denatured gel (polyacrylamide gel electrophoresis, PAGE), with a molecular weight of about 45 000. The results of further antigen-antibody analysis showed that the antigen/antibody hybridization reaction of the expression product with polyclonal or monoclonal antibody of JE virus (mosquito isolates, encephalitis isolates) and serum samples of patients with acute JE virus infection could be completely consistent. The recombinant product showed negative antigen/antibody hybridization reactions with mosquito-transmitted flaviviruses, such as dengue virus and yellow fever virus polyclonal antibodies, but positive reactions with polyclonal antibodies to West Nile virus and Murray Valley encephalitis virus. Conclusions　In this study, the recombinant expression of the NS1 protein of JE virus was successfully obtained, and the antigen/antibody reaction between the recombinant protein and samples of patients infected with mosquito-borne flavivirus and JE virus was analyzed. The study results provide important basic data for elucidating the antigen-antibody reaction between the NS1 protein of JE virus and mosquito-borne flavivirus. The recombinant expression protein obtained in this study provides an important material basis for further research on the function of JE virus NS1 protein.

OBJECTIVETo analyze the genotype of a patient suspected for thalassemia through a series of experiments.

METHODSConventional methods for detecting common thalassemia mutations was used in conjunction with multiplex ligation-dependent probe amplification (MLPA) in order to determine the genotype of the patient. Corresponding primers were designed for developing a Gap-PCR system for detecting rare type mutations.

RESULTSThe patient was identified as a homozygote for Chinese Gγ(Aγδβ)-thal deletion, with clinical manifestations tending to be intermediate or severe based on the hematological characteristics. A Gap-PCR system has been developed for detecting the above mutation with accuracy and rapidity.

CONCLUSIONThe Chinese Gγ(Aγδβ)-thal is prevalent in southern China, and caution should be taken to avoid misdiagnosis. The Gap-PCR system for detecting Chinese Gγ(Aγδβ)-thal is suitable for extended applications for its simplicity and rapidity.

OBJECTIVE: To develop a system for rapid detection of JAK2 V617F mutation among patients with myeloproliferative diseases. METHODS: Specific primers and TagMan probes were designed for the mutant and wild type alleles based on the principle of real-time PCR. A complete system including the method for detection and product for quality control were established through the evaluation of sensitivity and accuracy of the method, double-blind trial, and preparation of negative and positive controls through site-directed mutagenesis and molecular cloning. RESULTS: A system for rapid detection of the JAK V617F mutation has been developed. Compared with Sanger sequencing, the sensitivity and specificity of the method have both reached 100%. Meanwhile, 1000 normal samples and 1 case with the JAK2 V617F mutation were detected, which gave a population rate of 1‰. CONCLUSION The system was fast, accurate, cheap, high throughput, and easy to use. It can be utilized as a routine test. Although the JAK2 V617F mutation is rare in the population, it should be screened among myeloproliferative neoplasm patients.
Alleles ; DNA Mutational Analysis ; Double-Blind Method ; Humans ; Janus Kinase 2 ; genetics ; Mutation ; Myeloproliferative Disorders ; genetics ; Real-Time Polymerase Chain Reaction ; Sensitivity and Specificity

Objective: To investigate the clinicopathologic characteristics, immunophenotypes, and differential diagnostic features of extra-pleural solitary fibrous tumor (SFT) with uncommon histology. Methods: Seven cases of extra-pleural SFT with uncommon histology were collected during January 2015 and December 2016 in Zhejiang Provincal People′s Hospital; the clinical and radiologic features, histomorphology, immunophenotype and prognosis were analyzed. EnVision method was used for immunohistochemical staining of STAT6, CD34 and other differential diagnosis associated markers. Results: There were five male and two female patients, age from 23 to 54 years (mean=39 years). Three tumors were located in the soft tissue of head and neck, two in trunk subcutaneous soft tissue, one in sella region, and one in the kidney. Grossly the tumors ranged from 0.4 to 8.0 cm (mean=3.1 cm). Microscopically, all three head and neck cases resembled giant cell angiofibroma/giant cell subtype SFT, and one case showed sheet-like pattern of the multinucleated syncytial cells, creating a biphasic arrangement similar to myofibroma. Both truncal tumor resembled lipomatous type SFT, with one similar to dermatofibrosarcoma protuberans and the other to atypical spindle cell lipomatous tumor. The sella tumor showed morphology of a conventional SFT with high grade sarcomatous transformation. The renal tumor demonstrated a malignant SFT with entrapped benign renal tubules, mimicking a biphase synovial sarcoma or a malignant mixed epithelial and stromal tumor. By immunohistochemistry, all seven SFTs showed diffuse and strong nuclear reactivity to antibody against STAT6. Conclusions Extra-pleural SFTs show a significant heterogeneity of morphology and biological behavior which could cause differential confusion.Careful attention to its characteristic histomorphology with the use of STAT6 immunohistochemistry can help distinguish this tumor from its many mimickers.

Objective:To identify the gene mutations associated with facial cleft-related orbital hypertelorism in 3 pairs of monozygotic twins with different phenotypes (with/without hypertelorism) and to investigate their mechanisms.Methods:From May 2014 to May 2019, 3 pairs of monozygotic twins, 2 males and 4 females, aged 5-18 years, were treated in Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, one with normal orbital distance and widening of orbital distance was caused by facial fissure. Among the twins, there was 1 case of orbital hypertelorism and the other case of without orbital hypertelorism, and the hypertelorism was caused by facial cleft. To screen for mutations in hypertelorism, whole genome sequencing was performed on 3 pairs of twins. The Sanger method was used to sequence the exons of 33 patients with facial fissure associated hypertelorism and 50 healthy individuals in the same period to identify the genes selected by the whole genome sequencing. The periosteal tissues were obtained from patients and healthy people during plastic surgery. The cells were cultured, the activity of alkaline phosphatase was measured, and the osteogenic differentiation was identified by alizarin red staining, real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expression of signal transduction pathways in periosteal cells.Results:Whole genome sequencing analysis showed that in all three sets of twins, a new synonymous mutation (c.1479G>A, p. Q493Q) was found in the MAML3. In Sanger exon sequencing, 17(51.5%) of 33 patients with hypertelorism carried the mutation, while no mutation was detected in 50 normal controls. The result of periosteum-derived cytology showed that the expression of MAML3 mRNA and protein in the patient-derived cells was lower than that in the healthy-derived cells. Three, 7, 14 days after osteoinduction, the ALP activity in the cells from the patients was higher than that from the healthy subjects (8.540±1.450, 20.740±2.514, 24.090±3.213 vs. 5.268±0.482, 11.680±1.527, 13.200±0.592; all P<0.05). Fourteen days after osteoinduction, the result of alizarin red staining showed that there were more erythema formation in the cells from the patients than those from the healthy subjects, these result suggest that MAML3 mutation may lead to over-differentiation of human periosteal-derived cells. The mRNA and protein expression levels of hes1 and hes5 downstream of the Notch signal pathway were down-regulated in the periosteal cells of the patients, while Wnt3a and β-catenin mRNA and protein expression levels were up-regulated in the Wnt signal pathway. Conclusions:The MAML3 gene (c.1479G>A, p. Q493Q) mutation is one of the causative genes of facial cleft-related hypertelorism. Notch and Wnt/β-catenin signaling pathway play an important role in the pathogenesis of hypertelorism.