[Short-term efficacy and safety of cardiac contractility modulation in patients with heart failure].

Objective: To investigate the short-term efficacy and safety of cardiac contractility modulation (CCM) in patients with heart failure. Methods: This was a cross-sectional study of patients with heart failure who underwent CCM placement at the First Affiliated Hospital of Xinjiang Medical University from February to June 2022. With a follow-up of 3 months, CCM sensation, impedance, percent output, and work time were monitored, and patients were compared with pre-and 3-month postoperative left ventricular ejection fraction (LVEF) values, and 6-minute walk test distance and New York Heart Association (NYHA) cardiac function classification, and the occurrence of complications was recorded. Results: CCM was successfully implanted in all 9 patients. Seven(7/9) of them were male, aged (56±14) years, 3 patients had ischaemic cardiomyopathy and 6 patients had dilated cardiomyopathy. At 3-month postoperative follow-up, threshold was stable, sense was significantly lower at follow-up than before (right ventricle: (16.3±7.0) mV vs. (8.2±1.1) mV, P<0.05; local sense: (15.7±4.9) mV vs. (6.7±2.5) mV, P<0.05), and impedance was significantly lower at follow-up than before (right ventricle (846±179) Ω vs. (470±65) Ω, P<0.05, local sense: (832±246) Ω vs. (464±63) Ω, P<0.05). The CCM output percentage was (86.9±10.7) %, the output amplitude was (6.7±0.4) V, and the daily operating time was (8.6±1.0) h. LVEF was elevated compared to preoperative ((29.4±5.2) % vs. (38.3±4.3) %, P<0.05), the 6-minute walk test was significantly longer than before ((96.8±66.7)m vs. (289.3±121.7)m, P<0.05). No significant increase in the number of NYHA Class Ⅲ-Ⅳ patients was seen (7/9 vs. 2/9, P>0.05). The patient was not re-hospitalised for worsening heart failure symptoms, had no malignant arrhythmic events and experienced significant relief of symptoms such as chest tightness and shortness of breath. No postoperative complications related to pocket hematoma, pocket infection and rupture, electrode detachment, valve function impairment, pericardial effusion, or cardiac perforation were found. Conclusions: CCM has better short-term safety and efficacy in patients with heart failure.

[The efficacy and safety of laparoscopic radical gastrectomy after neoadjuvant chemotherapy combined with immunotherapy and targeted therapy].

Objectives: To explore the efficacy and safety of laparoscopic radical gastrectomy after neoadjuvant chemotherapy combined with immunotherapy and targeted therapy in patients with gastric cancer. Methods: A retrospective analysis of clinical and pathological data of 20 patients with locally advanced gastric cancer (clinical TNM stage T3-4aN+M0) admitted to the Cancer Hospital, Chinese Academy of Medical Sciences from July 2021 to July 2023. All patients received 3 cycles of SOX (Oxaliplatin+S-1) regimen combined with immunotherapy (Trastuzumab) and targeted therapy (Apatinib) as neoadjuvant treatment followed by laparoscopic radical gastrectomy for gastric cancer. Surgical outcomes, postoperative pathological response, and postoperative recovery were observed. Quantitative data, except for age and operation time, were expressed using Median (range). Results: Among the 20 patients, there were 18 males and 2 females, aged 41 to 73 years [(60.6±9.7) years]. All 20 patients underwent laparoscopic surgical treatment after neoadjuvant therapy, with one patient undergoing laparoscopic conversion to open total gastrectomy with partial transverse colon resection due to tumor invasion into the transverse mesocolon. Eight patients underwent totally laparoscopic radical gastrectomy, all with Billroth Ⅱ+Braun anastomosis at the distal stomach. Eleven patients underwent laparoscopic-assisted radical gastrectomy, among which total gastrectomy with Roux-en-Y anastomosis was performed in ten cases, and proximal gastrectomy with esophagogastrostomy overlap anastomosis was performed in one case. The mean operation time for the 20 patients was (165.0±34.1) minutes; intraoperative blood loss was 80 (20-100) ml; and the number of lymph nodes retrieved was 68 (21-89). Postoperative pathological TNM staging revealed stage T0N0M0 in six cases, stage Ⅰ in two cases, stage Ⅱ in three cases, and stage Ⅲ in nine cases. Six patients (30.0%) achieved pathological complete response, and nine patients (45.0%) achieved significant pathological response. The median postoperative time to flatus was 4 (1-5) days; oral intake resumed after 3 (2-5) days; and the median length of hospital stay was 13 (6-19) days. One patient developed colonic anastomotic leakage with intra-abdominal infection, and one patient developed duodenal stump leakage with intra-abdominal infection, both classified as Clavien-Dindo grade 3A complications, and improved after treatment and discharged. One patient developed gastric paresis, and two patients developed pleural effusion, classified as Clavien-Dindo grade 2 complications, and improved after treatment and discharged. There were no deaths within 30 days after discharge. Conclusions: Laparoscopic radical gastrectomy for gastric cancer after neoadjuvant treatment with the SOX regimen combined with immunotherapy and targeted therapy is safe and feasible, with satisfactory short-term efficacy. However, there is an increase in overall surgical risk and difficulty, and it is recommended to be performed in experienced gastric cancer centers.

Clinical outcomes of SMILE and WFG-LASIK used to treat myopia and astigmatism: A systematic review and meta-analysis.

PURPOSE: To evaluate the safety, efficacy and postoperative visual quality of small incision lenticule extraction (SMILE) and Wavefront-Guided Laser in situ keratomileusis (WFG-LASIK) and to analyze their efficacy in correcting astigmatism. METHODS: A systematic literature search was performed using Cochrane Collaboration methodology. Databases searched included PubMed, Embase, the Cochrane Library and Web of Science. RevMan software version 5.3.0 was used for meta-analysis. RESULTS: A total of 976 eyes were included in 8 studies, of which 539 eyes underwent SMILE and 437 eyes underwent WFG-LASIK. There were no statistically significant differences in the proportion of eyes achieving uncorrected distance visual acuity of 20/20 or better (P=0.18), the proportion of eyes within±0.50 diopter of target refraction postoperatively (P=0.10), or the postoperative magnitude of cylinder (P=0.10). Regarding the Alpins vector analysis of astigmatism, there was no statistically significant difference in the surgical magnitude of error (P=0.09) between the two groups. WFG-LASIK has a lower surgical angle of error (P= 0.002) and higher surgical correction index of cylinder (P=0.03) than SMILE. In terms of aberrations, higher order aberrations (P=0.46), spherical aberrations (P=0.22) and trefoil (P=0.56) were not statistically different, while WFG-LASIK induced less coma than SMILE surgery (P=0.02). CONCLUSION: Both SMILE and WFG-LASIK are safe and effective ways to correct myopia and astigmatism. Compared with SMILE, WFG-LASIK has a lower surgical angle of error, higher surgical correction index of cylinder and induces less coma.

[Ovarian Sertoli-Leydig cell tumors: DICER1 hotspot mutations and associated clinicopathological features].

Objective: To investigate DICER1 hotspot mutations in ovarian Sertoli-Leydig cell tumor (SLCT) and its associated clinicopathological features. Methods: Forty-three SLCTs and 40 other sex cord-stromal tumors (SCSTs) diagnosed between 2010 and 2017 at Fudan University Shanghai Cancer Center were examined for somatic DICER1 hotspot mutations by Sanger sequencing. The associations between mutation status and clinicopathological features, including patient age, tumor differentiation and recurrence, were analyzed. Results: Somatic DICER1 mutations were found in 51% (22/43) of SLCTs, while none in the other 40 SCSTs. The most common mutation of DICER1 was p.D1709N in exon 24 (41%, 9/22) and the second most common mutation of DICER1 was p.E1813K in exon 25 (14%, 3/22). A novel frameshift mutation (c.5464delG, p.M1837fs*16) was identified in one SLCT with microcystic pattern. Mutations were more likely to occur in patients under forty years of age (P=0.046), whereas no significant associations were found between DICER1 mutations and clinical symptoms, morphology or tumor recurrence. Conclusions: Somatic DCIER1 hotspot mutations are specifically found in SLCT and may serve as an ancillary marker in differential diagnosis of SLCT from other SCST. The mutations occur more often in young patients (<40 years old). Additional studies are warranted to examine the associations between DICER1 mutations and clinicopathological features and prognosis of SLCT.

[Pediatric colonoscopy findings and changing patterns from Beijing in one institutional experience over 12 years].

OBJECTIVE: To investigate the demographics and diagnostic yield in a cohort of Chinese pediatric patients undergoing colonoscopy in one institution over 12 years. METHODS: The study participants were consecutive patients aged <18 years that underwent their first colonoscopy in the endoscopy center at Peking University Third Hospital between Jan. 1, 2005 and Dec. 31, 2017. Demographic, endoscopic, and pathological findings were collected. According to the age of the patients, they were divided into 0-3 year-old group, 4-6 year-old group, 7-14 year-old group and 15-17 year-old group. The patients were also divided into 2005-2011 group and 2012-2017 group, according to the time of colonoscopy. RESULTS: The cohort consisted of 326 patients, including 205 boys (62.9%) and 121 girls (37.1%). In the study, 31 patients (9.5%) were in 0-3 year-old group, 28 (8.6%) were in 4-6 year-old group, 96 (29.4%) were in 7-14 year-old group and 171 (52.5%) in 15-17 year-old group. The terminal ileum intubation success rate was 90.5% (295/326). No serious complications such as hemorrhage or perforation occurred during the procedures. The cleaning effect was good in 92.3% (301/326) of the patients. A total of 204 patients (62.6%) received a positive diagnosis under colonoscopy. 27.0% (88/326) of the patients was diagnosed as nonspecific colitis or terminal ileitis. 46 (14.1%) with inflammatory bowel disease (IBD) and 39 (12.0%) with polyp. The diseases were significantly different among the different age groups. The highest IBD diagnostic rate was found in 0-3 year-old group (7/31, 22.5%), while the highest polyp finding rate was in 4-6 year-old group (8/28, 28.6%). The number of the patients in 0-3 year-old group was significantly increasing in 2012-2017 group compared with 2005-2011 group (27/191 vs. 4/135, P=0.001), while the terminal ileum intubation success rate was higher (179/191 vs. 116/135, P=0.037). However, comparisons between years 2005-2011 and 2012-2017 showed that neither IBD nor polyp detection rate changed significantly (P=0.850). CONCLUSION: Colonoscopy in pediatric patients was a safe and effective procedure. Colitis or terminal ileitis was the primary finding during colonoscopy while IBD was the second one, and polyp was the third. However, the diagnostic yield did not change significantly. IBD was not as quickly increased in our hospital as it was in South China.

Regulatory effects of CCDC3 on proliferation, migration, invasion and EMT of human cervical cancer cells.

OBJECTIVE: To elucidate the potential effects of Coiled coil domain-containing 3 (CCDC3) on proliferative, migratory, invasive potentials and epithelial-mesenchymal transition (EMT) of human cervical cancer cells. MATERIALS AND METHODS: Protein and mRNA levels of CCDC3 in C33 and HeLa cells were determined by quantitative Real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Proliferative capacity and clonality of C33 and HeLa cells transfected with sh-CCDC3 were evaluated by cell counting kit-8 (CCK-8) and colony formation assay, respectively. Transwell assay and wound healing assay were conducted to determine the invasive and migratory potentials of cervical cancer cells with CCDC3 knockdown. Protein expressions of EMT-related genes in C33 and HeLa cells with CCDC3 knockdown were determined by Western blot. RESULTS: Transfection of sh-CCDC3 in C33 and HeLa cells markedly inhibited CCDC3 expression compared with those transfected with sh-EGFP. CCDC3 knockdown remarkably attenuated proliferative, migratory and invasive capacities. Moreover, CCDC3 knockdown inhibited protein levels of EMT-related genes in C33 and HeLa cells. CONCLUSIONS: Low expression of CCDC3 attenuated proliferative, migratory, invasive potentials and EMT of cervical cancer cells. Hence, CCDC3 may be utilized as a novel therapeutic target for cervical cancer.

[Analysis of factors relevant to primary non-response to infliximab treatment in patients with Crohn's disease].

Objective: To evaluate factors relevant to primary non-response in patients with Crohn's disease undergoing infliximab (IFX) treatment at week 14. Methods: Patients with Crohn's disease in the Third Hospital of Peking University who were subject to IFX treatment more than 3 times and followed-up for more than 14 weeks from October 2015 to October 2018 were reviewed. The response was defined by a decrease of ≥100 points from baseline in the Crohn's Disease Activity Index. The clinical data and laboratory examinations of the two groups were compared, and the treatment outcomes of non-responders were also followed up. Results: A total of 41 patients were enrolled, among which 27(65.9%) were male. The median age at treatment was 25 years, and 8 patients lost primary response (19.5%). There was no significant difference in the sex, age at diagnosis or treatment, Montreal disease type and laboratory examinations [C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), hemoglobin, albumin] in the response group and non-response group at baseline phase (P>0.05). Baseline CRP decreased from 17.7 (26.2) mg/L to 2.2 (3.6) mg/L in the response group, but increased from 11.7 (9.5) mg/L to 31.6 (28.4) mg/L in the non-response group at week 14 (P=0.024). The trend of ESR change in the response group [from 23.0 (28.5) mm/h to 7.0 (8.5) mm/h] and the non-response group [from 24.5 (22.5) mm/h to 35.0 (26.5) mm/h] was similar with that of CRP (P=0.036). Hemoglobin and albumin were significantly elevated in the response group, but not in the non-response group at week 14 (P=0.593, P=0.255). Among the non-response patients, 5 were converted to responsive after the treatment protocols had been adjusted. The combined immunosuppressant treatment all obtained clinical response. Conclusions: The elevated CRP and/or ESR may serve as indicators of primary non-response (at week 14) to IFX treatment among Crohn's disease patients, and the combination of immunosuppressive agents may be one of the effective treatments after excluding infection and other causes.

Tau-mediated iron export prevents ferroptotic damage after ischemic stroke.

Functional failure of tau contributes to age-dependent, iron-mediated neurotoxicity, and as iron accumulates in ischemic stroke tissue, we hypothesized that tau failure may exaggerate ischemia-reperfusion-related toxicity. Indeed, unilateral, transient middle cerebral artery occlusion (MCAO) suppressed hemispheric tau and increased iron levels in young (3-month-old) mice and rats. Wild-type mice were protected by iron-targeted interventions: ceruloplasmin and amyloid precursor protein ectodomain, as well as ferroptosis inhibitors. At this age, tau-knockout mice did not express elevated brain iron and were protected against hemispheric reperfusion injury following MCAO, indicating that tau suppression may prevent ferroptosis. However, the accelerated age-dependent brain iron accumulation that occurs in tau-knockout mice at 12 months of age negated the protective benefit of tau suppression against MCAO-induced focal cerebral ischemia-reperfusion injury. The protective benefit of tau knockout was revived in older mice by iron-targeting interventions. These findings introduce tau-iron interaction as a pleiotropic modulator of ferroptosis and ischemic stroke outcome.

Glioma cells promote angiogenesis through the release of exosomes containing long non-coding RNA POU3F3.

OBJECTIVE: Angiogenesis is a key event in the progression of gliomas, and emerging evidence suggests that exosomes are signaling extracellular organelles that modulate the tumor microenvironment and promote angiogenesis and tumor progression. This study aimed to explore the mechanism by which glioma-derived exosomes affect angiogenesis. MATERIALS AND METHODS: qRT-PCR was used to determine the expression level of linc-POU3F3 in glioma tissue as well as glioma cell lines. Ultrafiltration combined with a purification method was used to isolate exosomes derived from A172 cells (A172-Exo) and linc-POU3F3 shRNA-treated A172 cells (shA172-Exo). Transmission electron microscopy, Western blot and tunable resistive pulse sensing (TRPS) were used to identify exosomes. In vitro migration, proliferation, and tube formation experiments, as well as in vivo CAM assays, were used to analyze the pro-angiogenesis ability of exosomes. qRT-PCR and Western blot were used to identify expression levels of angiogenesis-related genes and proteins in human brain microvascular endothelial cells (HBMECs) after being cultured with exosomes. RESULTS: The levels of linc-POU3F3 were upregulated in glioma tissue and significantly correlated with the advanced tumor stage. A172 cells exhibited the highest expression level. A172-Exo was similar to shA172-Exo (50-100 nm in diameter) and expressed Alix, Tsg101 and CD9, while the expression level of linc-POU3F3 in A172-Exo was significantly higher than that in shA172-Exo. HBMECs rapidly internalized A172-Exo and shA172-Exo, and the linc-POU3F3 expression level in HBMECs treated with A172-Exo was significantly higher than the level in HBMECs treated with shA172-Exo. A172-Exo exhibited better function in promoting HBMECs migration, proliferation, tubular-like structure formation in vitro and arteriole formation in vivo. The gene and protein expression level of bFGF, bFGFR, VEGFA, and Angio in HBMECs treated with A172-Exo was much higher than that of HBMECs treated with shA172-Exo. CONCLUSIONS: These results indicated that gliomas can induce angiogenesis by secreting exosomes enriched in linc-POU3F3. Exosomes and lncRNA-POU3F3 may, therefore, function as a putative therapeutic target in glioma.

[Impact of 2013 American Society of Clinical Oncology/College of American Pathologist guidelines on borderline immunostaining results for HER2: a retrospective study on HER2 FISH results in 1 780 cases of invasive breast cancers].

OBJECTIVE: To analyze the impact of the revised 2013 American Society of Clinical Oncology/College of American Pathologist(ASCO/CAP)HER2 testing guidelines on the status of HER2 and its clinical significance in invasive breast cancers by fluorescent in situ hybridization(FISH). METHODS: One thousand seven hundred and eighty invasive breast cancer cases with equivocal 2+ immunostaining detected by FISH were retrospectively selected from 2010 to 2014, and the HER2/CEP17 dual-probe results were evaluated according to both the 2007 and 2013 ASCO/CAP guidelines for comparative analysis. RESULTS: Among the 1 780 IHC HER2 (2+ ) invasive breast cancers, the number of HER2 positive, equivocal and negative case were 310(17.41%), 66(3.71%)and 1 404(78.88%) respectively, basing on the 2007 guidelines; whereas basing on the 2013 ASCO/CAP HER2 guidelines, the number of HER2 positive, equivocal and negative case was 360 (20.22%), 182 (10.23%)and 1 238 (69.55%) respectively. Compared with the 2007 guidelines, the proportion of positive and equivocal cases were higher in the 2013 guidelines (17.41% versus 20.22%, 3.71% versus 10.23% respectively), while the proportion of negative cases was lower(78.88% versus 69.55%). CONCLUSIONS: Using the 2013 ASCO/CAP guidelines could lead to an increase in positive and equivocal cases, and a decrease in negative cases. The increase can probably be attributable to the inclusion of HER2 copy number besides HER2/CEP17 ratio as positive criteria, and it improves the accuracy and may be of important value for screening more population who benefit from HER2 targeting treatment; however the benefits for HER2 positive with low HER2 copy number and the clinical significance of the equivocal cases need to be further investigated.

Clinical significance of hmgb1 expression in human gastric cancer.

High mobility group box 1 (HMGB1) has been proved to be implicated in a variety of cell physiological and pathological behaviors including immune response, inflammation and cancer. Accumulating evidence suggests that HMGB1 plays a critical role in the development and progression of multiple malignancies. However, the clinical significance and prognosis of HMGB1 expression in some cancers remain controversial. The present study aimed to investigate whether overexpression of HMGB1 is an independent prognostic factor in patients with gastric cancer. The correlation of HMGB1 expression with clinicopathologic characteristics and prognosis was assessed by immunohistochemical assay through tissue microarray procedure in 50 primary gastric cancer cases. Our results indicated that the positive expression of HMGB1 was significantly increased in the nucleus of gastric cancer tissues compared with the adjacent non-cancerous tissues (ANCT) (64.0% vs 44.0%, P=0.025), but was not linked to the clinicopathologic features, including the TNM stage (P=0.533) and metastatic lymph node (P=0.771), in patients with gastric cancer. Kapalan-Meier and log-rank analysis demonstrated that overexpression of HMGB1 did not exert significant impact on the overall survival of patients with gastric cancer (P=0.805). Furthermore, Cox regression analysis showed that high HMGB1 protein expression did not represent an independent risk factor for patients with gastric cancer (P=0.677). Taken together, our findings suggest that high expression of HMGB1 is not correlated with the clinicopathologic characteristics of gastric cancer, and cannot serve as an independent prognostic biomarker for patients with gastric cancer.

IL-10 treatment is associated with prohibitin expression in the Crohn's disease intestinal fibrosis mouse model.

Prohibitin, which can inhibit oxidative stress and mitochondrial dysfunction, has been shown to have significant anti-inflammatory activities. Here, we investigate the effects of altering prohibitin levels in affected tissues in the interleukin-10 knockout (IL-10KO) mouse model with intestinal fibrosis. The aim of this study is to investigate the effects of IL-10 on prohibitin and the role of prohibitin in intestinal fibrosis of murine colitis. After the mice were treated with IL-10, prohibitin expression and localization were evaluated in IL-10KO and wild-type (WT, 129/SvEv) mice. The colon tissue was then investigated and the potential pathogenic molecular mechanisms were further studied. Fluorescence-based quantitative polymerase chain reaction (FQ-PCR) and immunohistochemistry assays revealed a significant upregulation of prohibitin with IL-10 treatment. Furthermore, IL-10 decreases inflammatory cytokines and TGF-ß1 in the IL-10KO model of Crohn's disease and demonstrates a promising trend in decreasing tissue fibrosis. In conclusion, we hypothesize that IL-10 treatment is associated with increased prohibitin and would decrease inflammation and fibrosis in an animal model of Crohn's disease. Interestingly, prohibitin may be a potential target for intestinal fibrosis associated with inflammatory bowel disease (IBD).

Contralateral nasolabial flap for reconstruction of midface defects.

Reconstruction of defects in the midface near the nasolabial fold or the inner canthus of the eye can be a challenge for plastic surgeons. Many methods such as skin grafting and skin flaps have been proposed for reconstruction of these defects. This report presents the results of using a contralateral nasolabial flap with a pedicle containing the lateral nasal branch of the angular artery. A flap with a pedicle approximately 10 mm wide was transferred to the opposite side through a tunnel at the nasion near the inner canthus for reconstruction of defects with surface areas ranging from 5.3 to 31.0 cm(2) and depths ranging 2.2-6.5 mm. Since 2008, with careful design, eight patients with skin tumor excisions have obtained effective functional and aesthetic results. This method is a suitable surgical procedure for reconstruction of moderate to wide side defects in the special midface area of the nasal sidewall toward the medial canthus. Level of Evidence V This journal requires that authors assign a level of evidence to each article.

Dose estimation by chromosome aberration analysis and micronucleus assays in victims accidentally exposed to (60)Co radiation.

The objective of this study was to assess the radiation exposure levels in victims of a (60)Co radiation accident using chromosome aberration analysis and the micronucleus assay. Peripheral blood samples were collected from three victims exposed to (60)Co 10 days after the accident and were used for the chromosome aberration and micronucleus assays. After in vitro culture of the lymphocytes, the frequencies of dicentric chromosomes and rings (dic+r) and the numbers of cytokinesis blocking micronuclei (CBMN) in the first mitotic division were determined and used to estimate radiation dosimetry. The Poisson distribution of the frequency of dic+r in lymphocytes was used to assess the uniformity of the exposure to (60)Co radiation. Based on the frequency of dic+r in lymphocytes, estimates of radiation exposure of the three victims were 5.61 Gy (A), 2.48 Gy (B) and 2.68 Gy (C). The values were estimated based on the frequencies of CBMN, which were 5.45 Gy (A), 2.78 Gy (B) and 2.84 Gy (C). The estimated radiation dosimetry demonstrated a critical role in estimating the radiation dose and facilitating an accurate clinical diagnosis. Furthermore, the frequencies of dir+r in victims A and B deviated significantly from a normal Poisson distribution. Chromosome aberration analysis offers a reliable means for estimating biological exposure to radiation. In the present study, the micronucleus assay demonstrated a high correlation with the chromosome aberration analysis in determining the radiation dosimetry 10 days after radiation exposure.

Expression and quantification of LYVE-1 in human colorectal cancer.

The recent discovery of a new hyaluronan (HA) receptor, LYVE-1 (lymphatic vessel endothelial HA receptor), has been received with great interest regarding its specific expression in the lymphatic system. The process of lymphangiogenesis or the formation of new lymphatics in tumours is important because it serves as a major route for cancer metastasis. Therefore, methods to quantify lymphangiogenesis by measuring LYVE-1 have been studied extensively in searching for its possible role in cancer diagnosis, prognosis and even targeted treatment of lymphatic tumour metastasis. Here we report a quantitation study on lymphangiogenesis by either quantitative PCR or immunohistochemistry approaches in detecting LYVE-1 expression in human colorectal tumour. Real-time quantitative polymerase chain reaction (RTQ-PCR) was carried out to quantify LYVE-1 levels in colorectal cancer samples. Also, the same specimen was observed for LYVE-1 expression by immunohistochemical stain. By RTQ-PCR amplification, LYVE-1 was highly expressed in colorectal specimens and LYVE-1 signal from non-cancer tissue of normal control was much weaker by conventional RTPCR. Immunohistochemical stain showed that LYVE-1 was significantly expressed in cancer tissues (especially in the margin region of cancer), whereas in non-cancer specimens fewer positive stains were revealed. The results suggested that the LYVE-1 molecule was expressed significantly in colorectal specimens, which may imply a new marker for a malignant situation.

Dissecting the pet18 mutation in Saccharomyces cerevisiae: HTL1 encodes a 7-kDa polypeptide that interacts with components of the RSC complex.

The yeast pet18 mutant exhibits three distinct phenotypes: temperature-sensitive lethality, failure to maintain a dsRNA virus, and respiration deficiency. We have isolated a yeast mutant, H53, with phenotypes identical to those of pet18. Based on PCR and Southern hybridization analysis, H53 was found to result from a large chromosomal deletion extending from YCR019w to YCR028c on chromosome III. Genetic analysis was carried out on H53 to correlate individual loci with each of the observed phenotypes. Disruption of YCR020c-a/MAK31 brought about a loss of dsRNA without affecting the temperature sensitive phenotype. The loss of YCR020w-b/HTL1, which encodes a hypothetical protein of 78 amino acids in length, was shown to be responsible for the temperature-sensitive lethality of the H53 mutant. Using immunoblotting, we demonstrated that a 7-kDa protein was indeed expressed in wild-type yeast, but not in a HTL1 deletion mutant. Moreover, the significance of HTL1 was investigated by isolating genes that are functionally associated with HTL1. We demonstrated that Rsc8p interacts physically with Htl1p, and that the genes RSC3, STH1 and RSC30 interact with HTL1. Thus, HTL1 may play a role in the function of the RSC complex.

Deletion of mitochondrial ATPase inhibitor in the yeast Saccharomyces cerevisiae decreased cellular and mitochondrial ATP levels under non-nutritional conditions and induced a respiration-deficient cell-type.

T(1), a mutant yeast lacking three regulatory proteins of F(1)F(o)ATPase, namely ATPase inhibitor, 9K protein and 15K protein, grew on non-fermentable carbon source at the same rate as normal cells but was less viable when incubated in water. During the incubation, the cellular ATP content decreased rapidly in the T(1) cells but not in normal cells, and respiration-deficient cells appeared among the T(1) cells. The same mutation was also induced in D26 cells lacking only the ATPase inhibitor. Overexpression of the ATPase inhibitor in YC63 cells, which were derived from the D26 strain harboring an expression vector containing the gene of the ATPase inhibitor, prevented the decrease of cellular ATP level and the mutation. Isolated T(1) mitochondria exhibited ATP hydrolysis for maintenance of membrane potential when antimycin A was added to the mitochondrial suspension, while normal and YC63 mitochondria continued to show low hydrolytic activity and low membrane potential. Thus, it is likely that deletion of the ATPase inhibitor induces ATPase activity of F(1)F(o)ATPase to create a dispensable membrane potential under the non-nutritional conditions and that this depletes mitochondrial and cellular ATP. The depletion of mitochondrial ATP in turn leads to occurrence of aberrant DNA in mitochondria.

Binding of an intrinsic ATPase inhibitor to the F(1)FoATPase in phosphorylating conditions of yeast mitochondria.

Yeast mitochondrial ATP synthase has three regulatory proteins; ATPase inhibitor, 9K protein, and 15K protein. A mutant yeast lacking these three regulatory factors was constructed by gene disruption. Rates of ATP synthesis of both wild-type and the mutant yeast mitochondria decreased with decrease of respiration, while their membrane potential was maintained at 170-160 mV under various respiration rates. When mitochondrial respiration was blocked by antimycin A, the membrane potential of both types of mitochondria was maintained at about 160 mV by ATP hydrolysis. ATP hydrolyzing activity of F(1)FoATPase solubilized from normal mitochondria decreased in proportion to the rate of ATP synthesis, while the activity of the mutant F(1)FoATPase was constant regardless of changes in the rate of phosphorylation. These observations strongly suggest that F(1)FoATPase in the phosphorylating mitochondria is a mixture of two types of enzyme, phosphorylating and non-phosphorylating enzymes, whose ratio is determined by the rate of respiration and that the ATPase inhibitor binds preferentially to the non-phosphorylating enzyme.

Src activation in the induction of long-term potentiation in CA1 hippocampal neurons.

Long-term potentiation (LTP) is an activity-dependent strengthening of synaptic efficacy that is considered to be a model of learning and memory. Protein tyrosine phosphorylation is necessary to induce LTP. Here, induction of LTP in CA1 pyramidal cells of rats was prevented by blocking the tyrosine kinase Src, and Src activity was increased by stimulation producing LTP. Directly activating Src in the postsynaptic neuron enhanced excitatory synaptic responses, occluding LTP. Src-induced enhancement of alpha-amino-3-hydroxy-5-methylisoxazolepropionic acid (AMPA) receptor-mediated synaptic responses required raised intracellular Ca2+ and N-methyl-D-aspartate (NMDA) receptors. Thus, Src activation is necessary and sufficient for inducing LTP and may function by up-regulating NMDA receptors.

Arthroscopic debridement for osteoarthritis of the knee: a seven years follow-up study.

We reviewed 32 knees with osteoarthritis of the knee treated by either arthroscopic debridement in association with drilling the subchondral bone or arthroscopic debridement alone and followed for 2.5 to 11 years. Eighteen knees had arthroscopic debridement and drilling the subchondral bone, and 14 knees had arthroscopic debridement alone. In the group treated with arthroscopic debridement and drilling the subchondral bone, 55.6% had good to excellent results, 22.2% had fair results, and 22.2% had poor results. In the group that had arthroscopic debridement alone, 57.2% had good to excellent results, 35.7% had fair results, and 7.1% had poor results. There was better relief of pain in the group with arthroscopic debridement alone.