[The mechanism of circ_0023990/miR-873-5p/ANXA2 axis regulating radiosensitivity and development of thyroid carcinoma].

Objective: To explore the effect and possible mechanism of circ_0023990 on the radiosensitivity of thyroid cancer cells. Methods: qRT-PCR was used to detect the expression of circ_0023990 in the cancer tissues of 55 patients with thyroid cancer and thyroid cancer cell lines (TPC-1, KTC-1, FTC-133 and CAL-62), and the relationship between the expression of circ_0023990 in cancer tissues and the clinical characteristics of the patients were analyzed. Thyroid cancer cells TPC-1 and KTC-1 were divided into sh-circ_0023990 group, sh-NC group, sh-circ_0023990+anti-miR-873-5p group, sh-circ_0023990+anti-miR-NC group, miR-873-5p group, miR-NC group, miR-873-5p+pcDNA-ANXA2 group and miR-873-5p+pcDNA group, and then clone formation experiment was used to detect cell radiosensitivity. After each group of cells was irradiated with 4Gy radiation, the expression of γH2AX protein in the cells was detected by Western Blot. The dual luciferase reporter gene experiment verified the targeting relationship between circ_0023990 and miR-873-5p or miR-873-5p and ANXA2. Results: The expression of circ_0023990 in thyroid cancer tissues was higher than that in normal tissues (2.15±0.09 vs. 0.97±0.05, P<0.05), and its expression was closely related to tumor size, lymph node metastasis and TNM staging of patients with thyroid cancer (P<0.05). The expression of circ_0023990 in thyroid cancer cell lines (TPC-1, KTC-1, FTC-133 and CAL-62) were higher than that of normal thyroid cells HTori-3 (3.16±0.38, 2.63±0.28, 1.82±0.24, 1.71±0.22 vs. 1.00±0.10, all P<0.05). The survival scores of TPC-1 and KTC-1 cells in the sh-circ_0023990 group were significantly lower than those in the sh-NC group (P<0.05), and the sensitization ratios were 2.482, 1.643; The survival scores of TPC-1 and KTC-1 cells in the sh-circ_0023990+anti-miR-873-5p group were higher than those in the sh-circ_0023990+anti-miR-NC group (P<0.05), and the sensitization ratios were 0.305, 0.441, respectively. The survival scores of TPC-1 and KTC-1 cells in the miR-873-5p group were lower than those in the miR-NC group (P<0.05), and the sensitization ratios were 2.044, 1.653 respectively. The survival scores of TPC-1 and KTC-1 cells in the miR-873-5p+pcDNA-ANXA2 group was higher than that in the miR-873-5p+pcDNA group (P<0.05), and the sensitization ratios were 0.496, 0.686, respectively. The expression of γH2AX protein in TPC-1 and KTC-1 cells of the 4 Gy+sh-circ_0023990 group were higher than that in the 4 Gy+sh-NC group (2.68±0.27 vs. 1.87±0.25, 2.46±0.19 vs. 1.77±0.14; all P<0.05), but the expression of γH2AX protein in TPC-1 and KTC-1 cells of the 4 Gy+sh-circ_0023990+anti-miR-873-5p group were lower than that in the 4 Gy+sh-circ_0023990+anti-miR-NC group (1.13±0.09 vs. 1.69±0.09, 1.11±0.08 vs. 1.60±0.08; both P<0.05). The expression of γH2AX protein in TPC-1 and KTC-1 cells in the 4 Gy+miR-873-5p group were higher than that in the 4 Gy+miR-NC group (2.35±0.16 vs. 1.84±0.14, 2.26±0.12 vs. 1.77±0.13; both P<0.05), but the expression of γH2AX protein in TPC-1 and KTC-1 cells of the 4 Gy+miR-873-5p+pcDNA-ANXA2 group were lower than that in the 4 Gy+miR-873-5p+pcDNA group (1.96±0.12 vs. 2.41±0.12, 1.92±0.07 vs. 2.28±0.12; both P<0.05). circ_0023990 targeted the negative regulation of miR-873-5p, and ANXA2 was the target gene of miR-873-5p. Conclusion: circ_0023990 was highly expressed in thyroid cancer tissues and cell lines, and it may promote the radiotherapy resistance of thyroid cancer cells in vivo through regulating miR-873-5p/ANXA2 axis.

[Targeted therapy of pyogenic sterile arthritis, pyoderma gangrenosum, and acne syndrome (PAPA): a case report and literature review].

Objective: To analyze the clinical course and targeted therapy of pyogenic sterile arthritis, pyoderma gangrenosum, and acne (PAPA) syndrome. Methods: The clinical history of a 6-year-old boy with PAPA syndrome, who was admitted to Hong Kong University Shenzhen Hospital in September 2017, was reviewed. His genetic diagnosis was confirmed by whole exome sequencing. The response to targeted therapy was evaluated by comparing the inflammatory markers (erythrocyte sedimentation rate (ESR) and C reactive protein (CRP) and serum cytokines (interleukin (IL)-1, IL-6 and tumor necrosis factor-α (TNF-α)) before and after biological agents treatment. For literature review, "PAPA syndrome" and"PSTPIP1 gene"were used as keywords to retrieve papers published from January 1997 to December 2019 from Pubmed, Wanfang and CNKI database. Results: The patient was a 6-year-old boy, admitted to the hospital due to recurrent joint swelling and pain for more than 4 years. Before treatment, the CRP (256 mg/L), ESR (105 mm/1 h) and cytokines including serum TNF-α (7.43 ng/L), IL-1 (<5 ng/L), IL-6 (301 ng/L) were significantly elevated. Culture of the joint effusion was negative, but the IL-6 level was above 1 000 ng/L. MRI showed osteomyelitis at the lower end of the right femur. Gene detection found a heterozygous variation of PSTPIP1 gene (c.748G>A, p.E250K). Arthralgia once alleviated after the initiation of tocilizumab and infliximab, but recurred after 1 year of treatment. Thereafter, the anti-IL-1 receptor antagonist (Anakinra) was commenced, followed by a significant improvement of the arthralgia, and a complete remission during the follow-up. Besides, the level of CRP, ESR, serum TNF-α, IL-1 and IL-6 were all decreased to normal on the last followed up in December 2019. Literature review found 29 articles and 87 patients in total. The initial symptoms included those of arthritis (n=58), pyoderma gangrenosum (n=33), and acne (n=24). Among all the cases, 13 genotypes were confirmed, and 47 variations involved amino acid p.E250. Steroid and/or biological agents were used in most patients. Conclusions: PAPA syndrome should be suspected in children with recurrent pyogenic sterile arthritis, and an early diagnosis could be achieved by genetic test. Targeted treatment with biological agent may control the symptoms effectively. Biological agents can control symptoms of this disorder effectively.

LncRNA SNHG20 promoted the proliferation of glioma cells via sponging miR-4486 to regulate the MDM2-p53 pathway.

OBJECTIVE: Evidence demonstrated the critical role of long noncoding RNAs (lncRNAs) in the initiation and development of human cancers. LncRNA small nucleolar RNA host gene 20 (SNHG20) was recently reported to promote the progression of several cancers; however, the function of SNHG20 in glioma has not been characterized. PATIENTS AND METHODS: Differential expression of SNHG20 in glioma tissues and cell lines was analyzed by RT-qPCR. Cell Counting Kit-8 (CCK-8) assay was performed to detect the cell viability. The targets prediction was analyzed with the TargetScan database. Western blot was performed to check the protein expression. RESULTS: SNHG20 was overexpressed in glioma tissues and cell lines. Down-regulation of SNHG20 suppressed the proliferation, migration and induced apoptosis of glioma cells. Molecular studies uncovered that SNHG20 acted as a competing endogenous RNA (ceRNA) to sponge the expression of miR-4486. MiR-4486 was down-regulated in glioma tissues and significantly inversely correlated with the expression of SNHG20. Restoration of miR-4486 remarkably attenuated the promotion effect of SNHG20 on the growth of glioma cells. Further study revealed that miR-4486 targeted the E3 ubiquitin ligase mouse double minute 2 (MDM2) and negatively regulated the expression of MDM2. Down-regulation of MDM2 by miR-4486 increased the abundance of p53 in glioma cells. CONCLUSIONS: We identified the functional mechanism by which SNHG20 modulated the malignancy of glioma cells via targeting the miR-4486/MDM2/p53 pathway. Interrupting the expression of SNHG20 might be a novel strategy to suppress the progression of glioma.

Effect of downregulated lncRNA NBAT1 on the biological behavior of glioblastoma cells.

OBJECTIVE: To investigate the expression of lncRNA neuroblastoma associated transcript 1 (NBAT1) in human glioma cell lines and its underlying mechanism. Effect of NBAT1 on biological behaviors of T98 and U87 cells are also explored. PATIENTS AND METHODS: The mRNA expressions of NBAT1 in 48 cases of glioblastoma tissues and 30 cases of normal brain tissues were accessed by Real-time fluorescence quantitative PCR (RT-PCR). The relationship between mRNA expression of NBAT1 and tumor size, malignancy, and prognosis were analyzed. Effects of NBAT1 on the proliferation of glioblastoma T98 and U87 cells were determined by CCK-8 assay and colony formation assay, respectively. RESULTS: NBAT1 expressions in glioblastoma tissues were lower than those in normal brain tissues, which was negatively correlated with malignancy degree (p<0.01). Protein levels of Akt were decreased in T98 and U87 cells transfected with si-NBAT1. Meanwhile, proliferation abilities of T98 and U87 cells transfected with si-NBAT1 were significantly decreased as well (p<0.01), which were reversed by transfection of si-Akt. CONCLUSIONS: Upregulated NBAT1 inhibits proliferation of T98 and U87 cells via regulating Akt, indicating that NBAT1 may be related to the malignancy and prognosis of gliomas.

Importance of CD44 on umbilical cord mesenchymal stem cells for expansion of hematopoietic cells.

Human umbilical cord mesenchymal stem cells (hUCMSCs) have important functions on the expansion of hematopoietic stem cells (HSCs) through providing the essential microenvironment for hematopoiesis. In order to test whether CD44 on hUCMSCs could have a key function for the ability of hUCMSCs to expand human HSCs, the soluble anti—CD44 antibody was added to the co—cultures of hUCMSCs and cord blood (CB) CD34+ cells, which blocked the ability of hUCMSCs to expand CB CD34+ cells significantly. Long—term culture initiating cell (LTC—IC) assay revealed that the ability of multipotent differentiation of CB CD34+ cells co—cultured with CD44 knockdown hUCMSCs could only retain lasting at most for 5 weeks in vitro. In vivo assay, based on non—obese diabetic/severe combined immunodeficient disease (NOD/SCID) mice, revealed that the hematopoietic reconstitution potential of CB CD34+ cells co—cultured with CD44 knockdown hUCMSCs is significantly reduced. The hematopoietic supporting ability of hUCMSCs in vivo and in vitro is reduced upon the knockdown of CD44. CD44 has important functions on the ability of hUCMSCs to expand human HSCs in the cell— extrinsic control.

Primary spinal melanoma of extramedullary origin: a report of three cases and systematic review of the literature.

STUDY DESIGN: A review of the literature and three case reports. OBJECTIVES: Primary spinal melanoma (PSM) of extramedullary origin is a rare malignant condition with limited current literature in regards to its clinical course, magnetic resonance imaging (MRI) findings, treatment strategies and prognosis. We reported here three cases of PSM of extramedullary origin. SETTING: China, Guangzhou. METHODS: We report three cases of PSM of extramedullary origin. The clinical and radiological findings of these cases were retrospectively analyzed. RESULTS: The three cases were all of males aged 39, 47 and 76 years, respectively. The duration of their symptoms was 3 weeks, 2 months and 11 months respectively. The extramedullary tumors were all well-defined solitary tumors and were located at C4-5, L2-3 and T9-10, respectively. In one case, involvement of the intervertebral foramen was found. Preoperative MRI showed hyperintense T1W signals and hypointense T2W signals in all three cases and all tumors were clinically misdiagnosed as schwannomas. The patients received total or subtotal resection surgery without radiotherapy or chemotherapy. Patients were alive at 18 months, 27 months and 36 months postoperative follow-up, respectively. CONCLUSION: PSM of extramedullary origin is a rare malignant tumor that shows characteristic findings on MRI. Surgical resection is the preferred treatment strategy.

Meta-analysis of the differentially expressed colorectal cancer-related microRNA expression profiles.

OBJECTIVES: Unique microRNAs (miRNAs) have been identified in colorectal cancer in recent studies which can be used to accurately diagnose the presence of colorectal cancer and help predict disease recurrence. Differential expression of specific miRNAs in tissues or blood offers the prospect of their use in early detection and screening for colorectal cancer. However, the experiments under different environments would produce different results. The purpose of this study was to get a reliable result on differentially expressed miRNAs related to colorectal cancer by integrating different studies. MATERIALS AND METHODS: A meta-analysis was performed to review three miRNA microarray datasets from three published literatures that compared the microRNAs expression profiles in colorectal cancer tissues with those in normal colorectal tissues. The R VennDiagram package was applied to identify the overlapping miRNAs with differential expression among these three studies. RESULTS: A total of 175 differentially expressed miRNAs were reported in the three miRNA expression profiling studies that compared colorectal cancer tissues with normal tissues, of which 25 miRNAs were reported at least by two studies including 15 up-regulated miRNAs and 10 down-regulated miRNAs. Among the 25 miRNAs, 15 ones were differentially expressed between early stage colorectal cancer and normal tissues including 11 up-regulated miRNAs and 4 down-regulated miRNAs, of which hsa-miR-195 (down-regulated) and hsa-miR-20a (up-regulated) were shared by these three studies. CONCLUSIONS: The 15 differentially expressed miRNAs, especially hsa-miR-195 and hsa-miR-20a may be used as potential biomarkers for early detection and screening of colorectal cancer.

Cystic fibrosis transmembrane conductance regulator protein expression rate in healthy spermatozoa is not correlated with ovum fertilisation rate.

Our previous studies have shown that the cystic fibrosis transmembrane conductance regulator (CFTR) was important for capacitation and fertilisation in mouse, guinea pig and human spermatozoa. However, it is unclear whether CFTR is correlated with ovum fertilisation rate. The present study was to test the possible relationship between spermatozoa CFTR protein expression rate in healthy men and ovum fertilisation rate during in vitro fertilisation. Ninety-four couples for female factor infertility for IVF-ET treatments were retrospectively studied. All the patients were divided into three groups based on the fertilisation rate of ovum in vitro. It was performed to explore whether there were differences in sperm CFTR protein expression rate among the three groups and the relevance between CFTR protein expression rate and ovum fertilisation rate. Our study showed that there was no significant differences in sperm CFTR protein expression rate among the three groups (F = 0.614, P = 0.544), and the relevance between spermatozoa CFTR protein expression rate and ovum fertilisation rate was not significantly different (r = 0.013, P = 0.904). These results further suggest that CFTR protein expression rate in healthy men spermatozoa was not associated with ovum fertilisation rate and thus we cannot predict ovum fertilisation results by sperm CFTR protein expression rate.

Transplantation of a reverse facial artery pedicled submandibular gland in a dog.

This study was performed to evaluate the possibility of using reverse facial artery pedicled submandibular gland transfer for treating keratoconjunctivitis sicca. A reverse facial artery pedicled submandibular gland, including the duct, from a beagle dog was transplanted into the temporal region in the same animal. 99mTc pertechnetate scintigraphy was used to monitor graft viability, and the grafted glands were examined histologically 12 weeks after transplantation. Postoperative 99mTc pertechnetate scintigraphy demonstrated viable salivary gland tissue in the transplanted region. Histologically, some of the acinar cells in the graft had atrophied. The transplantation of a reverse facial artery pedicled submandibular gland was successful and may be a simple, reliable treatment for patients with keratoconjunctivitis sicca.

Expression and pathological relevance of inducible nitric oxide synthase in osteosarcoma of the jaws.

The aim of the present study was to examine the expression of inducible nitric oxide synthase (iNOS) in osteosarcoma of the jaw, and its relationship with tumour angiogenesis and clinicopathological characteristics. Streptavidin peroxidase immunohistochemical staining was used to detect the expression level of iNOS and CD34 in paraffin-embedded samples from 25 patients. Osteosarcoma of the jaw was associated with overexpression of iNOS, which correlated with tumour microvessel density (MVD). iNOS expression correlated with the size, pathological grade and clinical stage of the osteosarcoma, and also with clinicopathological characteristics such as primary occurrence or recurrence of tumours. There was no correlation with metastasis. iNOS may promote tumour angiogenesis in osteosarcoma of the jaw, and so may represent an important target in anti-tumour therapy.

Role of Arg163 in the N-glycosidase activity of neo-trichosanthin.

Three mutant crystals of neo-trichosanthin (n-TCS), R163K, R163H and R163Q, were obtained by the hanging drop vapor diffusion method. Structure determination indicated that there are no significant differences between the mutants and n-TCS except in the active pocket. All of them were also soaked in sodium citrate buffer (pH 4. 5) containing 20% KCl and 10 mg/ml AMP. Structure determination suggests that in the active pocket of the crystals of R163K and R163H, parallel to the aromatic ring of Tyr70, each mutant possesses an adenine. The relationship between structure and function is discussed. Biochemical analysis reveals that the mutants R163K and R163H have N-glycosidase activity, while R163Q does not. This suggests that R163 is a crucial residue for the enzyme activity of n-TCS, and its role is providing proton.

Role of TYR70 in the N-glycosidase activity of neo-trichosanthin.

Trichosanthin (TCS) is a type I ribosome-inactivating protein (RIP) which possesses rRNA N-glycosidase activity. TCS has long been used as an abortifacient in China. In recent years, its immunomodulatory, anti-tumor and anti-HIV properties have attracted more and more attention. An isoform of trichosanthin, neo-trichosanthin (n-TCS), has been cloned and expressed as recombinant protein. The biochemical studies revealed that n-TCS has virtually the same rRNA N-glycosidase activity as TCS. The crystal structure of n-TCS is similar to TCS. The crystal of Y70A n-TCS, the mutant of recombinant n-TCS, was soaked in sodium citrate buffer (pH 5.5) containing 25% KCl and AMP (10 mg/ml) prior to data collection. After structure determination and refinement, no electron density corresponding to adenine can be detected around the active pocket. Furthermore, the reaction products of Y70A n-TCS and AMP incubated at various reaction times were analyzed using HPLC. No adenine can be detected. These results suggest that Tyr70 is crucial to n-TCS for its substrate recognition, binding and perhaps N-glycosidase activity.

Limb-salvage for osteosarcoma.

Limb-salvage surgery plays an important role in the treatment of osteosarcoma. Among 104 patients with osteosarcoma we managed from 1974 to 1993, 60 were treated with limb-salvage procedure. 44 patients with IIB tumor received adjuvant chemotherapy, and the five-year survival rate was 46.69%. The local recurrence rate of these 60 patients was 15%, the infection rate 11.7%, and the prosthetic mechanical complication was 13.3%. Our results suggest that limb-salvage procedures are not only possible but also can provide patients with better function.

The value of electrophysiologic testing in patients resuscitated from documented ventricular fibrillation.

INTRODUCTION: Electrophysiologic testing is performed in patients resuscitated from ventricular fibrillation (VF) on the assumption that sustained monomorphic ventricular tachycardia (VT) may be a precursor to VF, with the former amenable to assessment by serial drug testing. METHODS AND RESULTS: We assessed the usefulness of this strategy by analyzing clinical and electrophysiologic data of 42 survivors (29 men and 13 women; mean age 54 +/- 14 years) of VF without a reversible cause. All patients had VF documented on ECG and required defibrillation. Underlying heart diseases included coronary disease in 28, dilated cardiomyopathy in 3, arrhythmogenic right ventricular dysplasia in 1, and no apparent structural heart disease in 10 patients. Only 2 (4.7%) patients had a prior history of documented VT. The electrophysiologic study was performed 7 to 30 days after VF. Programmed stimulation at the right ventricular apex using at least two drive cycle lengths and up to three extrastimuli induced sustained monomorphic VT in 4 (9.5%), sustained polymorphic VT in 3 (7.1%), nonsustained monomorphic VT in 1 (2.3%), nonsustained polymorphic VT in 5 (11.9%), and VF in 13 (30.9%) patients. Two patients with documented prior VT and coronary disease had sustained VT induced during the electrophysiologic study. On the other hand, sustained monomorphic VT was induced in 53 of the 59 (90%) patients (45 men and 14 women; mean age 57 +/- 16 years) with clinically documented VT concurrently studied using the same stimulation protocol. CONCLUSION: We conclude that reproducible induction of sustained monomorphic VT in survivors of documented VF is uncommon. It may be more cost effective to proceed directly to treatment with implantable cardioverter defibrillators in these patients.

Effect of adenosine or adenosine triphosphate on antidromic tachycardia.

OBJECTIVES: This study was designed to determine the effect of adenosine or adenosine triphosphate (ATP) on antidromic tachycardia. BACKGROUND: Adenosine and adenosine triphosphate are useful for differential diagnosis of wide QRS tachycardia. It has been believed that tachycardia termination caused by these agents is due to the preferential depressive effect on the atrioventricular (AV) node, whereas their effect on accessory pathways is minimal. METHODS: We studied the effect of adenosine or ATP on the termination pattern of antidromic tachycardia in 17 patients (10 men, 7 women; mean age [+/- SD] 32 +/- 11 years) with one or more accessory pathways. Adenosine (6 to 12 mg [n = 10]) or ATP (8 to 20 mg [n = 7]) was injected rapidly through a central venous line and followed by 10 ml of saline flush after induction of sustained antidromic tachycardia. RESULTS: Tachycardia was terminated in < 2 min in 14 patients (82%) after the injection and remained unchanged in 3 (18%). Tachycardia termination was due to conduction block in the accessory pathway (anterograde limb) in seven patients (50%) and in the AV node (retrograde limb) in another seven. Adenosine or ATP caused accessory pathway block in seven (88%) of the eight patients lacking retrograde accessory pathway conduction and in none of the nine patients having retrograde accessory pathway conduction (p < 0.01). All five patients with an atriofascicular accessory pathway and unidirectional anterograde conduction had tachycardia termination due to anterograde accessory pathway block after injection of adenosine or ATP. CONCLUSIONS: 1) Adenosine or ATP effectively terminates antidromic tachycardia; 2) the termination is related to block in either the accessory pathway or the AV node; 3) accessory pathway block occurs in patients with a unidirectional, anterogradely conducting accessory pathway, especially an atriofascicular accessory pathway.

[Fast neutron treatment of osteosarcoma].

Twelve patients with advanced osteosarcoma were treated with fast neutron at the Neutron Therapy Facility, the Hight-Energy Physics Institute, Chinese Academy of Science. Of the 12 patients, 5 were operated after radiotherapy. Three of them received fast neutron alone, and the other 2 with added 60Co. The doses ranged from 660 NcGy to 2 168 NcGy. Pain was relieved and the tumor size decreased in all the treated patients. Only one case was found histologically free of viable tumor cells. Skin reaction was serious and the knee joint function was impaired in all patients.

[Detection of RB gene in osteosarcoma by PCR].

We have established a stable PCR system and studied the structural anomalies of RB gene by it in osteosarcoma, previous studies have proved that 13-17 exons of RB gene are the deletion hotpoints. We designed a pair of perimers, RB1, RB2 stretching across 14-16 exons and studied 10 cases of osteosarcoma. No structural anomaly was found in the leucocyte. Two cases (20%) showed no PCR product. This result demonstrates for the first time the deletion of RB gene is present in Chinese osteosarcoma patients.