Clinicopathological analysis of EWSR1/FUS::NFATC2 rearranged sarcoma in the left forearm: A case report.

BACKGROUND: We present a case of an EWSR1/FUS::NFATC2 rearranged sarcoma in the left forearm and analyze its clinicopathological and molecular features. CASE SUMMARY: The patient is a 23-year-old woman. Microscopically, the tumor cells were medium-sized round cells arranged in small nests. The cytoplasm was clear, nuclei were relatively uniform, chromatin was dense, nucleoli were visible, and mitotic figures were rare. Immunohistochemically, the tumor cells were positive for Vimentin, INI-1, CD99, NKX2.2, CyclinD1, friend leukaemia virus integration 1, and NKX3.1. Next-generation sequencing revealed the presence of the EWSR1-NFATC2 fusion gene. EWSR1/FUS::NFATC2 rearranged sarcomas are rare and can easily be misdiagnosed. CONCLUSION: Clinical imaging, immunohistochemistry, and molecular pathology should be considered to confirm the diagnosis.

Norcrassin A, a novel C16 tetranorditerpenoid, and bicrotonol A, an unusual dimeric labdane-type diterpenoid, from the roots of Croton crassifolius.

A novel C16 tetranorditerpenoid, norcrassin A (1), and an unusual dimeric labdane-type diterpenoid, bicrotonol A (2), were isolated from the roots of Croton crassifolius. Norcrassin A (1) featured a new carbon skeleton with an unprecedented 5/5/5/6 tetracyclic system. Bicrotonol A (2) possessed an unusual tetrahydroxypyran ring linkage connecting two labdane diterpenoid monomers. The structures of all compounds, including the absolute configuration, were elucidated by the interpretation of their NMR spectroscopic data, high resolution mass spectrometry, and single-crystal X-ray diffraction. A plausible biosynthetic pathway of 1 is proposed. The anti-Alzheimer's Disease (AD) activities of 1 and 2 are also evaluated using the AD pathological model.

Cytotoxic and antibacterial activities of iridoids and sesquiterpenoids from Valeriana jatamansi.

Three new iridoids, valejatanins A-C (1-3), and one new natrual iridoid (4), together with four known sesquiterpenoids (5-8), were isolated from the roots of Valeriana jatamansi Jones. Compounds 3 and 4 are C(4)-epimers. Their structures were elucidated by extensive spectroscopic analysis (IR, HRESIMS, 1D and 2D NMR) and by comparison of their NMR data with those of related compounds. The absolute configuration of 5 was determined for the first time by single-crystal X-ray diffraction analysis with Cu-Kα irradiation. The cytotoxic activities of all compounds were evaluated against HT29, K562 and B16 cancer cell lines in vitro by MTT assay. Valejatanin A (1) showed noteworthy cytotoxic activities with IC50 values of 22.17, 15.26, 3.53µg/mL against three cancer cell lines. The antibacterial activities of all compounds against bacteria were tested in vitro. Compound 6 exhibited antibacterial activities against Staphylococcus aureus and Pseudomonas aeruginosa.

Eremophilane-type Sesquiterpenes from the Leaves of Ligularia virgaurea.

A phytochemical study of the ethanolic extract of the leaves of Ligularia virgaurea led to the isolation of a new eremophilane-type sesquiterpene lactone, (4S,5R,6S,8S,lR)-6ß-angeloyloxy-eremophil-7(l l)-en-10ßH-8α,12-olide (1), along with a known eremophilane-type sesquiterpene, (4S,5R,6S,lOS)-6ß- angeloyloxy-10ßH-furanoeremophil-9-one (2). Their structures were elucidated by extensive spectroscopic methods, including ID and 2D nuclear magnetic resonance and high-resolution electrospray ionization mass spectrometry experiments, and the absolute configurations were confirmed by single-crystal X-ray diffraction analysis using the anomalous scattering of Cu Ka radiation.

A New Succinate Derivative from Ajuga decumbens.

A new succinate derivative, ethyl (5-formylfuran-2-yl)methyl succinate (1), along with three known compounds (2-4) have been isolated from the whole plants of Ajuga decumbens Thunb. Their structures were elucidated by extensive spectroscopic (1D and 2D NMR) and HR-ESI-MS data analysis, and literature values. Compound 1 was isolated as a new succinate derivative, and compounds 2 and 3 were for the first time separated from A. decumbens.

[Water extract from Codonopsis thalictrifolia wall affects the reproductive system of male infant rats].

OBJECTIVE: To study the impact of the water extract from Codonopsis thalictrifolia Wall (CTW) on the reproductive METHODS: We divided 32 male SD infant rats into four groups of equal number to be treated intragastrical-system of male infant rats. ly with distilled water (control) and CTW at 10 g/kg (low dose) , 20 g/kg (medium dose), and 40 g/kg (high dose), respectively, twice a day for 2 weeks. Then we killed the rats, measured the levels of testosterone (T), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in the serum, obtained the testis weight, body weight, testis visceral coefficient and sperm concentration, and detected sperm viability, sperm motility and the level of cyclic adenosine monophosphate (cAMP) in the Leydig cells, followed by RESULTS: Compared with the control group, the low-dose, me-analysis of differences among different groups using the SPSS software. Medium-dose and high-dose CTW groups showed significant decreases in the serum T level ([3.09 +/-0.42] vs [1.22 +/-0. 32] , [1.06 +/- 0.29] and [0.57 +/-0.18] nmol/L, P<0.01), testis weight ([1.40 +/-0.16] vs [0.96 +/-0.09], [0.92 +/-0.11] and [0.91 +/- 0.08] g, P <0.01), and sperm concentration ([1.03 +/-0.16] vs [0.19 +/-0.07], [0.17 +/-0.08] and [0.16 +/-0.07] x 10(6)/ml, P <0.01), but a dramatic elevation in the testis visceral coefficient ([42.22 +/- 3.02] vs [51.39 +/- 3.09], [52.28 +/- 4.86] and [54.13 +/-6.06] mg/10 g, P <0.01); the medium- and high-dose CTW groups exhibited remarkable increases in the levels of serum LH ([13.62+/-0.89] vs [14.69 +/-0.12] and [14.93 +/-0.28] ng/L, P<0.01) and FSH ([4.32 +/-0.18] vs [4.77 +/-0.23] and [4.89 +/-0. 38] IU/L, P <0.05); all the three CTW groups showed markedly inhibited serum T secretion ([1.85 +/- 0.18] vs [1.42 +/-0.15], [1.12+/-0.18] and [0.88 +/-0.21] nmol/L, P<0.01) and intracellular cAMP ([5.51 +/-0.12] vs [4.39+/-0.06], [4.28 +/-0.07] and [4.11 +/- 0.10] nmol/L, P <0.01) in the Leydig cells. CONCLUSION: The water extract from CTW may reduce the synthesis of testosterone in the serum of male infant rats through the PKA pathway and consequently inhibit their testicular development and sperm production and affect the development of their reproductive system.

[Bufo gargarizans mcl-1 cloning and its prokaryotic recombinant protein expression].

MCL-1 is encoded by myeloid cell leukemia-1 gene (mcl-1), which is one of the anti-apoptotic members of bcl-2 cell apoptotic gene superfamily. ChanSu is made of dorsal secretions of several Bufo species, commonly used in the prescriptions of traditional Chinese medicine for treating many diseases including cancer. To clarify if mcl-1 is expressed in the dorsal skin of B. gargarizans, the PCR (polymerase chain reaction) was performed with its dorsal skin first strand cDNA as the template and a pair of specific primers of mcl-1, and PCR products were cloned into the pGM-T vector. DNA sequencing indicated that the ORF length was 639 bp encoding 212 amino acid residues, and the homology of 44%-95% with the MCL-1 of several other animals. For the further studies on MCL-1 biological functions during the oncogenesis and preparation of its antibody, the prokaryotic expression construct of pET-28b-mcl-1 was prepared which was confirmed by DNA sequencing, and its recombinant protein expression (0.02% wet weight) in E. coli BL21 (DE3) strain was confirmed by SDS-PAGE and Western blotting.

Modification of chitosan membrane with poly(vinyl alcohol) and biocompatibility evaluation.

This work aimed to overcome chitosan (CS) membrane' drawbacks: mainly stiffness and hydrophobic surface by adding poly(vinyl alcohol) (PVA) and evaluate their biocompatibility. The chemical structure, crystalline and thermal properties were studied by FT-IR, XRD and DSC. The mechanical properties and wettability of CS/PVA membranes were studied by tensile test and static contact angle measurement. In vitro biocompatibility was also evaluated by MTS cytotoxicity assay and SEM examination. The results suggest that adding PVA into CS membrane could greatly improve CS membrane's flexibility and wettability. All the membranes prepared were biocompatible and have potential applications in GTR technology.

A facile approach to construct hybrid multi-shell calcium phosphate gene particles.

The calcium phosphate (CaP) particles have attracted much attention in gene therapy. How to construct stable gene particles was the determining factor. In this study, hybrid multi-shell CaP gene particles were successfully constructed. First, CaP nanoparticles served as a core and were coated with DNA for colloidal stabilization. The xi-potential of DNA-coated CaP nanoparticles was -15 mV. Then polyethylenimine (PEI) was added and adsorbed outside of the DNA layer due to the electrostatic attraction. The xi-potential of hybrid multi-shell CaP particles was slightly positive. With addition of PEI, the hybrid multi-shell particles could condense DNA effectively, which was determined by ethidium bromide (EtBr) exclusion assay. The hybrid particles were spherical and uniform with diameters of about 150 nm at proper conditions. By simple modification of PEI, the hybrid multi-shell CaP gene particles were successfully constructed. They may have great potential in gene therapy.

Preventive effect of gelatinizedly-modified chitosan film on peritoneal adhesion of different types.

AIM: To comparatively study the preventive effect of gelatinizedly-modified chitosan film on peritoneal adhesions induced by four different factors in rats. METHODS: Chitosan was chemically modified by gelatinization, and made into films of 60 microm in thickness, and sterilized. Two hundred Sprague-Dawley rats were randomly divided into five groups, Sham-operation group (group A), wound-induced adhesion group (group B), purified talc-induced adhesion group (group C), vascular ligation-induced adhesion group (group D), and infection-induced adhesion group (group E), respectively. In each group, the rats were treated with different adhesion-inducing methods at the cecum of vermiform processes and then were divided into control and experimental subgroups. Serous membrane surface of vermiform processes were covered with the films in the experimental subgroups, and no films were used in the control subgroups. After 2 and 4 wk of treatments, the abdominal cavities were reopened and the adhesive severity was graded blindly according to Bhatia's method. The cecum of vermiform processes were resected for hydroxyproline (OHP) measurement and pathological examination. RESULTS: Adhesion severity and OHP level: After 2 and 4 wk of the treatments, in the experimental subgroups, the adhesions were significantly lighter and the OHP levels were significantly lower than those of the control subgroups in group B (2 wk: 0.199 +/- 0.026 vs 0.285 +/- 0.041 microg/mg pr, P < 0.001; 4 wk: 0.183 +/- 0.034 vs 0.276 +/- 0.03 microg/mg pr, P < 0.001), D (2 wk: 0.216 +/- 0.036 vs 0.274 +/- 0.040 microg/mg pr, P = 0.004; 4 wk: 0.211 +/- 0.044 vs 0.281 +/- 0.047 microg/mg pr, P = 0.003) and E (2 wk: 0.259 +/- 0.039 vs 0.371 +/- 0.040 microg/mg pr, P < 0.001; 4 wk: 0.242 +/- 0.045 vs 0.355 +/- 0.029 microg/mg pr, P < 0.001), but there were no significant differences in groups A (2 wk: 0.141 +/- 0.028 vs 0.137 +/- 0.026 microg/mg pr, P = 0.737; 4 wk: 0.132 +/- 0.031 vs 0.150 +/- 0.035 microg/mg pr, P = 0.225) and C (2 wk: 0.395 +/- 0.044 vs 0.378 +/- 0.043 microg/mg pr, P = 0.387; 4 wk: 0.370 +/- 0.032 vs 0.367 +/- 0.041 microg/mg pr, P = 0.853); Pathological changes: In group B, the main pathological changes were fibroplasias in the treated serous membrane surface and in group D, the fibroplasia was shown in the whole layer of the vermiform processes. In group E, the main pathological changes were acute and chronic suppurative inflammatory reactions. These changes were lighter in the experimental subgroups than those in the control subgroups in the three groups. In group C, the main changes were foreign body giant cell and granuloma reactions and fibroplasias in different degrees, with no apparent differences between the experimental and control subgroups. CONCLUSION: The gelatinizedly-modified chitosan film is effective on preventing peritoneal adhesions induced by wound, ischemia and infection, but the effect is not apparent in foreign body-induced adhesion.

[Characteristics of genesis and development of peritoneal adhesion by different causes: experiment with rats].

OBJECTIVE: To investigate the characteristics of genesis and development of peritoneal adhesion by different causes. METHODS: 236 rats underwent laparotomy with their vermiform processes lifted up and were randomly divided into 5 groups: Group A (control group), with the vermiform process exposed to air for 5 min, Group B, with the vermiform process smeared with talcum powder; Group C, with the vermiform process scraped by scalpel; Group D, with the tip of vermiform process stabbed by needle so as to squeeze the contents of intestine to cause infection; and Group E, with the artery of vermiform process ligated. Then the abdominal incision was sutured. 1, 2, 4, and 6 weeks after the treatment 11-12 rats from each group were randomly to undergo laparotomy. The degree of adhesion was graded blindly by Bhatia's method. The vermiform process was resected to undergo pathological examination and examination of the level of organ hydroxyproline (OHP) was detected. RESULTS: (1) At different time points the adhesive grades of Groups B-E were all significantly higher than that of Group A (all P < 0.05) and the adhesive grades of Groups B and D were both significantly higher than those of Groups C and E (both P < 0.05). There were no significant differences in the adhesive degree 1, 2, and 4 weeks after the treatment between Groups C and E, however, the adhesive degree of Group E was significantly lower than that of Group C (P < 0.05). (2) There were not significant differences in the OHP levels at any time points in Group A (all P > 0.05). There were not significant differences in the OHP levels 1, 2, and 4 weeks after the treatment (all P > 0.05), and the levels 8 weeks after the treatment were all significantly lower than those 1, 2, and 4 weeks after the treatment (all P < 0.05) in Groups B, D, and E. In Group C the OHP level 2 weeks after the treatment was 0.275 +/- 0.031 microg/mg protein, significantly lower than that 1 week after (0.221 +/- 0.036 microg/mg protein, P < 0.05), and the OHP level 8 weeks after the treatment was 0.254 +/- 0.039 microg/mg protein, significantly lower than those 1, 2, and 4 weeks after (all P < 0.05). The OH levels 1, 2, and 4 weeks after the treatment of the 4 experimental groups were all significantly higher than that of the control group (all P < 0.05). 8 weeks after the treatment the level of OHP of Groups B was significantly higher than that of Group A (P < 0.05), however, the OHP levels of Group C, D, and E had all decreased to almost similar to that of Group A (all P > 0.05). (3) The adhesive degrees of Groups C and E were significantly positively correlated with the OHP level (both P < 0.05), however, the adhesive degrees of Groups B and D were not significantly correlated with the OHP level (both P > 0.05). The adhesive degrees 1, 2, and 4 weeks after the treatment of the 5 groups were all significantly positively correlated with the OHP levels (all P < 0.05, however, the adhesive degrees 8 weeks after the treatment of the 5 groups were all not significantly correlated with the OHP levels (all P > 0.05). (4) The main pathological changes of Group B were foreign body granuloma reaction and fibroplasia in Group B and unspecific inflammatory reaction and fibroplasia in Groups C, D, and E. CONCLUSION: Abdominal adhesions resulting from different causes show different characteristics. The abdominal adhesion caused by foreign bodies and that caused by infection are relatively severe and more difficult to recover than those caused by injury and ischemia. It is more reliable to use OHP level as a marker of abdominal adhesion in the early stage.

[Application of McAbs against rabies nucleocapsid in diagnosis of rabies street virus].

OBJECTIVE: McAbs against rabies nucleocapsid were used to detect rabies street viruses in animal brain specimens with indirect immunofluorescent assay to evaluate the sensitivity and specificity of this assay. METHODS: 62 specimen from rabid animal brains including genotype 1 to 7 and 271 specimens from different normal animal brains collected in Pasteur Institute in 2003 were tested and compared, using indirect immunofluorescent assay. All these specimens were identified and compared using rapid rabies enzyme immunodiagnosis, fluorescent antibody test and rabies virus isolation assay in neuroblastoma cell culture which were all provided by Pasteur Institute. RESULTS: Both sensitivity and the specificity of the indirect immunofluorescent assay were 100%. CONCLUSION: The results showed a positive of rabies virus detection with these methods.