Identification of Foramen Ovale With H-Figure Fluoroscopic Landmark Improves Treatment Outcomes in Idiopathic Trigeminal Neuralgia.

BACKGROUND: Because it is traditionally difficult and time-consuming to identify the foramen ovale (FO) with fluoroscopy, we recently developed the H-figure method to acquire fluoroscopic view of FO with shorter procedure time and less radiation. However, the impact of such an H-figure approach on the clinical outcomes of trigeminal ganglion radiofrequency thermocoagulation (RFT) in treating idiopathic trigeminal neuralgia (ITN) remains unclear. METHODS: In a 12-month follow-up retrospective cohort study, patients with ITN had fluoroscopy-guided RFT of trigeminal ganglion via either classic approach (n = 100) or H-figure approach (n = 136) to identify FO. Data of continuous variables were analyzed with a Shapiro-Wilk test for normality and subsequently with a Mann-Whitney test, and the binary data were analyzed with a χ 2 test. The primary outcome was the facial pain measured by a Visual Analog Scale (VAS) 1 year after the treatment. The secondary outcomes included the quality of the fluoroscopic FO views, the threshold voltage to provoke paresthesia, the procedure time, the number of fluoroscopic images, and the facial numbness VAS. RESULTS: Compared with the classic approach group, the H-figure approach group was associated with better long-term pain relief after the procedure, with significantly fewer patients had pain 3 months (6.6% vs 17.0%, P = .012) and 12 months (21.3% vs 38.0%, P = .005) after the procedure, and among patients who had pain after the procedure, patients in the H-figure group had significantly less pain 6 months after the procedure (VAS median [interquartile range (IQR)]: 3 [2-6] vs 6 [4-7], P < .001). Moreover, compared to the classic approach, the H-figure approach provided better fluoroscopic view of FO, lower threshold voltage to elicit paresthesia (median [IQR]: 0.2 [0.2-0.3] vs 0.4 [0.4-0.5] V, P < .0001), with shorter procedure time (median [IQR]: 7.5 [6.0-9.0] vs 14.0 [10.0-18.0] min, P < .0001), and required fewer fluoroscopic images (median [IQR]: 4.0 [3.0-5.0] vs 8.0 [6.0-10.0], P < .0001). CONCLUSIONS: RFT of the trigeminal ganglion using the H-figure approach is associated with superior longer term clinical pain relief than the classic approach in treating ITN.

Preprocedure ultrasound imaging combined with palpation technique in epidural labor analgesia.

BACKGROUND: For parturients with paroxysmal uterine contraction pain, rapid analgesia is needed. We used preprocedure ultrasound imaging combined with the palpation technique in epidural analgesia for labor, and evaluated the usefulness of this technique in epidural labor analgesia. AIM: To evaluate the usefulness of preprocedure ultrasound imaging in epidural analgesia for labor. METHODS: In this prospective randomized observational study, 72 parturients were assigned to two groups (combined or palpation group). The target interspace of all parturients was first identified by the palpation technique. Then in the combined group, preprocedure ultrasound imaging was used before epidural puncture. In the palpation group, only the traditional anatomical landmarks technique (palpation technique) was performed. The primary outcome was total duration of the epidural procedure (for the ultrasound group, the duration of the preprocedure ultrasound imaging was included). The secondary outcomes were the number of skin punctures, the success rate at first needle pass, the number of needle passes, the depth from the skin to epidural space, and the complications of the procedure. RESULTS: Total duration of the epidural procedure was similar between the two groups (406.5 ± 92.15 s in the combined group and 380.03 ± 128.2 s in the palpation group; P = 0.318). A significant improvement was demonstrated for epidural puncture and catheterization in the combined group. The number of needle passes was 1.14 in the combined group and 1.72 in the palpation group (P = 0.001). The number of skin puncture sites was 1.20 in the combined group and 1.25 in the palpation group (P = 0.398). The success rate at first needle pass was 88.89% in the combined group and 66.67% in the palpation group (P = 0.045). CONCLUSION: This study demonstrated that the total duration of epidural procedures with preprocedure ultrasound imaging combined with the palpation technique was not longer than the traditional anatomical landmarks technique, which were performed by six experienced anesthesiologists in parturients with normal weights undergoing labor analgesia.

An interobserver reproducibility analysis of size-set semiautomatic counting for Ki67 assessment in breast cancer.

PURPOSE: The proliferation marker Ki67 has prognostic and predictive values in breast cancer, and the cutoff of the Ki67 label index (LI) is a key index for chemotherapy. However, poor interobserver consistency in Ki67 assessment has limited the clinical use of Ki67, especially in luminal cancers. Here, we reported a modified Ki67 assessment method, size-set semiautomatic counting (SSSAC) and investigated its interobserver reproducibility. METHODS: One hundred invasive breast cancer tissues were set immunostained for Ki67 in one laboratory, scanned as digital slides, and sent to 41 pathologists at the laboratories of 16 hospitals for Ki67 LI assessment using size-set semiautomatic counting (SSSAC), size-set visual assessment (SSVA) and size-set digital image analysis (SSDIA) with a specific image viewing software (Aperio Image Scope, Leica, Germany). The intraclass correlation coefficient (ICC) and Bland-Altman plot were used to evaluate interobserver reproducibility. The Wilcoxon signed-rank test was used to analyze the difference in the Ki67 values assessed by SSSAC and SSDIA. RESULTS: SSSAC demonstrated better interobserver reproducibility (ICC = 0.942) than SSVA (ICC = 0.802). The interobserver reproducibility was better in Ki67 homogeneously stained slides and centralized hot-spot slides than in scattered hot-spot slides. The Ki67 value assessed with SSSAC was obviously higher than that assessed with SSDIA (negative ranks (SSDIA < SSSAC): N = 80, sum of ranks = 4274.50; positive ranks (SSDIA > SSSAC): N = 17, sum of ranks = 478.50; Z = -6.837; P < 0.001). CONCLUSION: SSSAC shows satisfactory interobserver reproducibility in the Ki67 assessment of breast cancer and may be a candidate standard method for Ki67 LI assessment in breast cancer and other malignancies.

The effect of ultrasound-guided percutaneous ozone injection around cervical dorsal root ganglion in zoster-associated pain: a retrospective study.

OBJECTIVE: This study was to evaluate the effectiveness of ultrasound-guided percutaneous ozone injections around the cervical dorsal root ganglions of zoster-associated pain (ZAP) patients. STUDY DESIGN: Retrospective comparative study. SETTINGS: The study was conducted at a pain center of a university hospital. PATIENTS AND METHODS: From June 2016 to July 2017, a total number of 30 patients with ZAP were treated with ultrasound-guided percutaneous ozone injection around the cervical dorsal root ganglion (DRG) at the injured nerve level (C2-C8). A volume of 3 mL ozone-oxygen mixture at a concentration of 30 µg/mL was injected into the area around the DRG. Patients were divided into two groups according to their disease duration: group A (at or <3 months) and group B (>3 months). The pain severity was assessed according to a visual analog scale, and imaging changes were evaluated by ultrasound. Patient improvements in pain and neurologic function were evaluated during a follow-up period from 1 to 3 months. RESULTS: The data showed that ozone injections reduced pain in patients with ZAP. However, the success rate of group A was higher than group B. After the injection, the von Frey data demonstrated decreases in both groups, but, there were no significant differences between the groups. Moreover, univariate logistic regression analysis and multivariate regression analysis showed a history of diabetes mellitus had a significant effect on the treatment results. CONCLUSIONS: Percutaneous ozone injection around the DRG might be a useful method for treatment-resistant cases of ZAP at the cervical level. Institutional Review Board (IRB) approval number: HK2017-1130.

Computed tomography-guided percutaneous ozone injection of the Gasserian ganglion for the treatment of trigeminal neuralgia.

OBJECTIVE: The aim of this study was to evaluate the therapeutic effect of computed tomography (CT)-guided percutaneous ozone injection for refractory trigeminal neuralgia. DESIGN: A retrospective evaluation was performed in the study. SETTING: The study was conducted at a university hospital pain center. PATIENTS AND METHODS: A total of 29 patients with a clinical diagnosis of refractory trigeminal neuralgia were enrolled. All patients were treated with a percutaneous ozone injection and one patient was excluded. There were 21 patients with classical trigeminal neuralgia (group A) and seven patients with painful trigeminal neuropathy caused by post-herpetic neuralgia (group B). The percutaneous injection was an oxygen-ozone mixture at an ozone concentration of 30 µg/mL into the Gasserian ganglion performed under CT guidance. The number of procedures performed varied from one to as many as 16. Outcomes were evaluated using visual analog scale (VAS) pain scores. RESULTS: The combined VAS scores were 7.11 ± 1.23 pretreatment, 2.86 ± 1.69 posttreatment (P < 0.05) and 3.25 ± 2.01 after 6-month follow-up (P < 0.05). In group A, the VAS scores were 7.10 ± 1.04 pretreatment and 2.90 ± 1.84 posttreatment (P < 0.05). In group B, the VAS scores were 7.14 ± 1.77 pretreatment and 2.71 ± 1.25 posttreatment (P < 0.05). After 6-months follow-up, the VAS score was 3.38 ± 2.18 in group A and 2.86 ± 1.46 in group B, a decrease compared to pretreatment (P < 0.05). VAS of Group A and B showed no difference not only in pretreatment but also in postreatment and follow-up. CONCLUSION: Percutaneous ozone injection is a safe and effective treatment for patients with refractory trigeminal neuralgia.

The antitumor efficacy of a novel adenovirus-mediated anti-p21Ras single chain fragment variable antibody on human cancers in vitro and in vivo.

Activated ras genes are found in a large number of human tumors, and therefore are one of important targets for cancer therapy. This study investigated the antitumor effects of a novel single chain fragment variable antibody (scFv) against ras protein, p21Ras. The anti-p21Ras scFv gene was constructed by phage display library from hybridoma KGHR1, and then subcloned into replication-defective adenovirus vector to obtain recombinant adenovirus KGHV100. Human tumor cell lines with high expression of p21Ras SW480, MDA-MB­231, OVCAR-3, BEL-7402, as well as tumor cell line with low expression of p21Ras, SKOV3, were employed to investigate antitumor effects in vitro and in vivo. Fluorescence microscopy demonstrated that KGHV100 was able to express intracellularly anti-p21Ras scFv antibody in cultured tumor cells and in transplantation tumor cells. MTT, Transwell, colony formation, and flow cytometry analysis showed that KGHV100 led to significant growth arrest in tumor cells with high p21Ras expression, and induced G0/G1 cell cycle arrest in the studied tumor cell lines. In vivo, KGHV100 significantly inhibited tumor growth following intratumoral injection, and the survival rates of the mice were higher than the control group. These results indicate that the adenovirus-mediated intracellular expression of the novel anti-p21Ras scFv exerted strong antitumoral effects, and may be a potential method for therapy of cancers with p21Ras overexpression.

Mutual regulation between CHD5 and EZH2 in hepatocellular carcinoma.

Chromodomain helicase DNA binding protein 5 (CHD5) acts as a tumor suppressor in many cancers. In the present study, we demonstrated that reduced levels of CHD5 in hepatocellular carcinoma (HCC) tissues were significantly associated with metastasis and poor prognosis. Gain-of-function assays revealed that CHD5 suppressed motility and invasion of HCC cells. Subsequent investigations showed that CHD5 was epigenetically silenced by polycomb repressive complex 2 (PRC2)-mediated the trimethylation of histone H3 at lysine 27 (H3K27me3) in HCC cells. Furthermore, overexpression of CHD5 repressed enhancer of zeste homolog 2 (EZH2) and activated PRC2 target genes, such as p16 and p21. Chromatin immunoprecipitation and luciferase reporter assays also showed that CHD5 and EZH2 bind to each other's promoters and inhibit transcription. These findings uncovered, for the first time, a mutual suppression regulation between CHD5 and EZH2, which may provide new insights into their potential therapeutic significance for HCC.

Integrated analysis of gene expression and DNA methylation changes induced by hepatocyte growth factor in human hepatocytes.

Hepatocellular carcinoma (HCC) is the one of most common malignant tumors. The tumor microenvironment has a role in not only supporting growth and survival of tumor cells, but also triggering tumor recurrence and metastasis. Hepatocyte growth factor (HGF), one of the important growth factors in the tumor microenvironment, has an important role in angiogenesis, tumorigenesis and regeneration. However, the exact mechanism by which HGF regulates HCC initiation and development via epigenetic reprogramming has remained elusive. The present study focused on the epigenetic modification and target tumor-suppressive genes of HGF treatment in HCC. Expression profiling and DNA methylation array were performed to investigate the function of HGF and examine global genomic DNA methylation changes, respectively. Integrated analysis of gene expression and DNA methylation revealed potential tumor suppressor genes (TSGs) in HCC. The present study showed the multiple functions of HGF in tumorous and non­tumorous pathways and global genomic DNA methylation changes. HGF treatment upregulated the expression of DNA methyltransferase 1 (DNMT1). Overexpression of DNMT1 in HCC patients correlated with the malignant potential and poor prognosis of HCC. Furthermore, integration analysis of gene expression and DNA methylation changes revealed novel potential tumor suppressor genes TSGs including MYOCD, PANX2 and LHX9. The present study has provided mechanistic insight into epigenetic repression of TSGs through HGF­induced DNA hypermethylation.

The antitumor effects of oncolytic adenovirus H101 against lung cancer.

Lung cancer is the leading cause of cancer mortality in both men and women, with dismal survival rates due to late-stage diagnoses and a lack of efficacious therapies. The new treatment options with completely novel mechanism of therapeutic activity are needed for lung cancer to improve patient outcome. The present study was aimed at testing the efficacy of recombinant adenovirus H101 as an oncolytic agent for killing human lung cancer cell lines in vitro and in vivo. We assessed the coxsackievirus adenovirus receptor (CAR) expression on human lung cancer cell lines by RT-PCR and immunocytochemistry staining. Viral infectivity and viral replication in lung cancer cells was assayed by flow cytometry and real-time fluorescent quantitative PCR. After H101 treatment, cytotoxic effect, cell cycle progression and apoptosis were further examined by lactate dehydrogenase release assay and flow cytometry in vitro, respectively. In vivo, antitumor effects of H101 were assessed on SCID Beige mice xenografted with human lung cancer cells. Receptor characterization confirmed that human lung cancer cell lines expressed CAR receptor for adenovirus type 5. Lung cancer cells were sensitive to infection by the H101 virus. H101 infection and replication resulted in very potent cytotoxicity, G2/M phase arrest and cell lysis. In vivo, we also showed that H101 significantly inhibited tumor growth following intratumoral injection, with virus replication, cell degeneration and necrosis in the tumor tissue. These results have important implications for the treatment of human lung cancer.

Caffeic acid phenethyl ester attenuates pro-inflammatory and fibrogenic phenotypes of LPS-stimulated hepatic stellate cells through the inhibition of NF-κB signaling.

Hepatic stellate cells (HSCs) are the major cell type involved in liver fibrosis. Lipopolysaccharide (LPS)-mediated signaling through Τoll-like receptor 4 (TLR4) in HSCs has been identified as a key event in liver fibrosis, and as the molecular link between inflammation and liver fibrosis. In this study, we investigated the effects of caffeic acid phenethyl ester (CAPE), one of the main medicinal components of propolis, on the pro-inflammatory and fibrogenic phenotypes of LPS-stimulated HSCs. HSCs from rats were isolated and cultured in Dulbecco's modified Eagle's medium (DMEM). Following treatment with LPS, HSCs showed a strong pro-inflammatory phenotype with an upregulation of pro-inflammatory mediators, and a fibrogenic phenotype with enhanced collagen synthesis, mediated by transforming growth factor-ß1 (TGF-ß1). CAPE significantly and dose-dependently reduced LPS-induced nitrite production, as well as the transcription and protein synthesis of monocyte chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6) and inducible nitric oxide synthase (iNOS), as determined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), western blotting and enzyme-linked immunosorbent assays (ELISA). CAPE further reduced the TGF-ß1-induced transcription and translation (protein synthesis) of the gene coding for collagen type I α1 (col1A1), in LPS-stimulated HSCs. Following LPS stimulation, the phosphorylation of the nuclear factor-κB (NF-κB) inhibitor IκBα and consequently, the nuclear translocation of NF-κB, were markedly increased in the HSCs, and these changes were reversed by pre-treatment with CAPE. In conclusion, CAPE attenuates the pro-inflammatory phenotype of LPS-stimulated HSCs, as well as the LPS-induced sensitization of HSCs to fibrogenic cytokines by inhibiting NF-κB signaling. Our results provide new insight into the treatment of hepatic fibrosis through regulation of the TLR4 signaling pathway.

[The quantitative detection of NKG2D, perforin and granzyme B gene expressions in lymphocytes by SYBR green I real-time fluorescence quantitative PCR].

AIM: To develop a SYBR Green I real-time fluorescence quantitative PCR for the detection of lymphocyte immune function. METHODS: The primers of NKG2D, perforin, granzyme B and keeping-home gene GAPDH were designed and synthesized according to NCBI gene sequences, and the real-time fluorescence quantitative PCR was established. The gene expressions of NKG2D, perforin and granzyme B in lymphocytes from cancer patients and CIK induced from the cancer patients lymphocytes in vitro were quantified by real-time fluorescence quantitative PCR. RESULTS: NKG2D, perforin and granzyme B mRNA could be specifically amplified and quantitatively detected by the SYBR Green I real-time fluorescence quantitative PCR according to agarose gel electrophoresis and melt curve analysis. The mRNA expression of granzyme B was reduced in lymphocytes from cancer patients, however the mRNA expressions of perforin and granzyme B were increased in CIK induced by cytokines and monoclone antibody compared with their lymphocytes(P<0.01). CONCLUSION: The SYBR Green I real-time fluorescence quantitative PCR is a useful method for the quantitative detection of NKG2D, perforin and granzyme B mRNA to investigate cellular immune function.

Effect of caffeic acid phenethyl ester on proliferation and apoptosis of hepatic stellate cells in vitro.

AIM: To investigate the role of nuclear factor-kappaB (NF-kappaB) inhibitor caffeic acid phenethy1 ester (CAPE) in the proliferation, collagen synthesis and apoptosis of hepatic stellate cells (HSCs) of rats. METHODS: The HSCs from rats were isolated and cultured in Dulbecco's Modified Eagle's Medium (DMEM) and treated with CAPE. The proliferation and collagen synthesis of HSCs were determined by (3)H-TdR and (3)H-proline incorporation respectively, and the expression of type I, III procollagen genes was further explored by in situ hybridization. Apoptosis cell indices (AIs) were examined using terminal deoxynucleotidyl transferase- mediated DIG-dUTP nick end labeling (TUNEL). RESULTS: In activated HSC in culture, CAPE significantly inhibited (3)H-TdR and (3)H-proline incorporation by HSCs at concentrations of 5 micromol/L and 10 micromol/L respectively. CAPE also reduced the type I procollagen gene expression (P<0.05) at higher concentration. Apoptosis of HSC was induced by CAPE and the AIs were time-and dose-dependently increased from 2.82+/-0.73 % to 7.66+/-1.25 % at 12 h (P<0.01) and from 3.15+/-0.88 % to 10.61+/-2.88 % at 24 h (P<0.01). CONCLUSION: CAPE inhibits proliferation and collagen synthesis of HSC at lower concentration and induces HSC apoptosis at higher concentration.

[Zymography identification of the matrix metalloproteinases synthesized by cultured hepatic stellate cells].

AIM: To establish a method for identification of the activities of matrix metalloproteinases (MMPs) synthesized by cultured hepatic stellate cells (HSC). METHODS: SDS-gelatin polyacrylamide gels electrophoresis was used for the identification of MMPs synthesized by cultured HSC. RESULTS: MMP-2 and MMP-9 in culture medium and HSC were sensitively identified, and MMP-2 activity was higher than MMP-9 in culture medium, the changes of MMPs in culture medium and HSC were also observed after 24h medicine treatment. CONCLUSION: Zymography is suitable for the studies on the role of HSC in the regulation of extracellular matrix turnover.