[The application of the diagnosis and therapy of single cell sequencing technology in central nervous system tumors].

Central nervous system (CNS) tumors have complex causes and poor prognosis. Tumor heterogeneity is a major cause of treatment failure, and in-depth understanding of the biodiversity of CNS tumors is critical. Single-cell sequencing technology provides an opportunity to reveal the complex ecosystem of CNS tumors. In this study, we review the significance of single-cell sequencing in exploring the heterogeneity, diagnosis, treatment, and prognosis of CNS tumors from three aspects: tumor stem cells, tumor microenvironment, and cerebrospinal fluid. Although most of the findings have not been clinically applicable, they lay the foundation for the development of new guidelines for CNS tumors that help improve tumor prognosis and prevent tumor recurrence.

[A sericin hydrogel scaffold for sustained dexamethasone release modulates macrophage polarization to promote mandibular bone defect repair in rats].

OBJECTIVE: To evaluate the efficacy of a modified sericin hydrogel scaffold loaded with dexamethasone (SMH-CD/DEX) scaffold for promoting bone defect healing by stimulating anti-inflammatory macrophage polarization. METHODS: The light-curable SMH-CD/DEX scaffold was prepared using dexamethasone-loaded NH2-ß-cyclodextrin (NH2-ß-CD) and sericin hydrogel and characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), biocompatibility assessment and drug release test. THP-1 macrophages incubated with the scaffold were examined for protein expressions of iNOS and Arg-1, mRNA expressions of IL-6, Il-10, Arg-1 and iNOS, and surface markers CD86 and CD206 using Western blotting, RT-qPCR, and flow cytometry. In a co-culture system of human periodontal ligament stem cells (HPDLSCs) and THP-1 macrophages, the osteogenic ability of the stem cells incubated with the scaffold was evaluated by detecting protein expressions of COL1A1 and Runx2 and expressions of ALP, Runx2, OCN and BMP2 mRNA, ALP staining, and alizarin red staining. In a rat model of mandibular bone defect, the osteogenic effect of the scaffold was assessed by observing bone regeneration using micro-CT and histopathological staining. RESULTS: In THP-1 macrophages, incubation with SMH-CD/DEX scaffold significantly enhanced protein expressions of Arg-1 and mRNA expressions of IL-10 and Arg-1 and lowered iNOS protein expression and IL-6 and iNOS mRNA expressions. In the co-culture system, SMH-CD/DEX effectively increased the protein expressions of COL1A1 and Runx2 and mRNA expressions of ALP and BMP2 in HPDLSCs and promoted their osteogenic differentiation. In the rat models, implantation of SMH-CD/DEX scaffold significantly promoted bone repair and bone regeneration in the bone defect. CONCLUSION: The SMH-CD/DEX scaffold capable of sustained dexamethasone release promotes osteogenic differentiation of stem cells and bone defect repair in rats by regulating M2 polarization.

[Myocardial protection of del Nido cardioplegia in adult cardiac and major vascular surgery with long aortic cross-clamp time].

Objective: To explore the safety and myocardial protection efficacy of del Nido cardioplegia in adult cardiac and major vascular surgery with long aortic cross-clamp (ACC) time. Methods: A total of 2 536 patients who underwent adult cardiac and major vascular surgery with ACC time>90 min at Beijing Anzhen Hospital from March 2018 to March 2023 were collected. The patients were divided into two groups according to the type of cardioplegia solution: the del Nido cardioplegia solution group (DC group) and the cold blood cardioplegia solution group (BC group). Preoperative baseline data of the patients (age, gender, comorbidities, ejection fraction, etc) were adjusted using propensity score matching (PSM). Cardiopulmonary bypass (CPB) time, ACC time, total amount of cardioplegia solution, in-hospital mortality rate, length of intensive care unit (ICU) stay, mechanical ventilation time, postoperative complications, left ventricular ejection fraction, and troponin levels were compared between the two groups. Results: After PSM, a total of 306 patients were included, including 223 males and 83 females, with a mean age of (52.0±12.3) years. There were 153 cases in the DC group and 153 cases in the BC group. Compared with the DC group, the cross-clamp time was longer [109(100, 150) min vs 102(91, 133) min, P<0.001], the rate of return to spontaneous rhythm was lower [51.6% (79/153) vs 86.9%(133/153), P<0.001], and intraoperative peak glucose was higher [12.6 (6.5, 15.9) mmol/L vs 10.1 (8.5, 12.4) mmol/L, P=0.005] in the BC group. In addition, perioperative mortality [4.6% (7/153) vs 3.3% (5/153), P=0.132], stroke[3.9% (6/153) vs 3.3% (5/153), P=0.759], renal insufficiency [3.3% (5/153) vs 6.5% (10/153), P=0.186], atrial fibrillation [4.6% (7/153) vs 2.6% (4/153), P=0.652] and low cardiac output syndrome [3.9% (6/153) vs 4.6% (7/153), P=0.716] did not differ between the two groups. Compared with BC group, DC group had lower level of high sensitivity troponin (hsTnI) [1.2 (0.8, 1.8) µg/L vs 1.3 (0.9, 2.3) µg/L, P=0.030] and creatine kinase isoenzyme (CK-MB) [31.0 (20.0, 48.9) µg/L vs 37.0 (24.0, 58.9) µg/L, P=0.011] at 24 h postoperatively, and shorter length of ICU stay [35.6 (19.8, 60.5) h vs 42.6 (21.9, 83.6) h, P=0.015] and mechanical ventilation time [20.5 (15.5, 41.0) h vs 31.5 (17.1, 56.0) h, P=0.012]. Subgroup analysis showed that in the 120-180 minute subgroup, patients in the DC group had a shorter cross-clamp time [132 (124, 135) min vs 136 (124, 138) min, P<0.001], while levels of hsTnI [1.6 (1.1, 2.0) µg/L vs 1.4 (1.0, 2.6) µg/L, P=0.030] and CK-MB [38.8 (23.5, 55.5) µg/L vs 37.0 (24.5, 62.3) µg/L, P=0.011] were higher than those in the BC group. Conclusions: In adult cardiac and major vascular surgery with ACC times>90 min, comparable myocardial protection is observed with the use of DC compared with BC. Additional advantages in glycemic control, return to spontaneous rhythm, and improved surgical procedures make DN an attractive alternative for myocardial protection in adult cardiac surgery.

[Pollution characteristics, source analysis, and risk assessment of metal and metalloid in PM2.5 of Shijiazhuang city].

In 2020, the mass concentration of PM2.5 in Shijiazhuang urban area was(80.30±71.43)µg/m3. The Spearman correlation analysis between metals and metalloids showed that Sb with Cd, Pb, Ni, Se, Cd with Pb, Ni, Se, Pb with Ni, Se, Ni with Se, and Se with Tl were positively correlated, with a coefficient greater than 0.5. The main sources of metals and metalloids of PM2.5 were traffic emissions, fuel combustion, metal smelting and dust. The HQ values of Pb, Hg and Mn for each population were less than 1, with lower non-carcinogenic risk. The R values of carcinogenic risk of Ni and Cd in each population were less than 1×10-6, which could be acceptable risk level for the population. The R values of carcinogenic risk of As and Cr in different populations were between 1×10-6 and 1×10-4, with potential carcinogenic risk, particularly higher in adult males.

[Application of intravenous sedation in 2 582 cases of oral and maxillofacial surgery].

OBJECTIVE: To analyze the clinical data of patients undergoing intravenous sedation in oral and maxillofacial surgery, to understand the epidemiological characteristics, to evaluate the efficacy and safety of intravenous sedation for oral surgery, and to summarize our experience. METHODS: We retrospectively reviewed the clinical data of patients undergoing intravenous sedation between January 2010 and December 2018 in the Department of Oral and Maxillofacial Surgery, Peking University School of Stomatology. The gender, age, source, disease types, the values of perioperative vital signs, the use of sedatives and analgesics, duration of surgery and sedation, effect of sedation during the operation and the postoperative anterograde amnesia were analyzed. RESULTS: A total of 2 582 patients experienced oral surgery by intravenous sedation. The peak age was 3.5 to 10 years and between 21 to 40 years. Supernumerary teeth (38%, 981/2 582) and impacted third molars (30%, 775/2 582) were the major disease types, and other types of disease accounted for 32 percent (826/2 582). The values of heart rate(HR), mean arterial pressure(MAP), respiration rate(RR) and bispectral index(BIS) showed statistically significant differences at the time of before sedation, local anesthesia injection, surgical incision, 10 min after operation and the end of operation. In the study, 69%(1 781/2 582) cases received midazolam alone, 7%(181/2 582) cases received propofol alone, and 24% (620/2 582) cases received midazolam and propofol combined for intravenous sedation. Fentanyl (33%, 852/2 582)was the most common intravenous analgesic we used, followed by flurbiprofen axetil (23%, 594/2 582) and ketorolac tromethamine (6%, 157/2 582). Besides, 35% (907/2 582) patients didn't use any intravenous analgesic during the surgery. The average operation time was (31.2±20.8) min, and the average sedation time was (38.4±19.2) min. During the surgery procedure, most of the patients scored on a scale of 2 to 4 according to the Ramsay sedation score (RSS). The postoperative anterograde amnesia rates of local anesthesia injection, surgical incision and dental drill during surgery were 94% (2 431/2 582), 92% (2 375/2 582) and 75% (1 452/1 936). CONCLUSION: Intravenous sedation on the oral and maxillofacial surgery is effective and safe, can make the patients more comfortable, and should be further promoted and applied.

Methylation and expression analysis of mismatch repair genes in extramammary Paget's disease.

BACKGROUND: Extramammary Paget's disease (EMPD) is a rare skin cancer with relative high frequencies of germline and somatic mismatch repair (MMR) genes mutations. However, the methylation and expression of these genes have not been validated in EMPD. OBJECTIVE: This study aims to confirm the methylation and expression of MMR genes in EMPD. METHODS: Immunohistochemical (IHC) staining detection and Methylation-specific PCR (MSP) were used to analyse MLH1, MSH2, MSH6 and PMS2 proteins' expression and promoters' methylation in 57 EMMD samples, and pyro-sequence was used to find highly methylated CpG sites in MSH2 promoter. RESULTS: Immunohistochemical detection displayed reduced expression of MSH2 in 38.6% EMPD cases but normal expression of MLH1, MSH6 and PMS2 in all tumour tissues. Hypermethylation also was found in the promoter of MSH2 but not in other MMR genes. Pyrosequencing of MSH2 promoter showed CpG6 (-87) and CpG3 (-98) were the most common two methylated CpG dinucleotides. There is a significant correlation between reduced MSH2 expression and MSH2 methylation. CONCLUSION: Reduced MSH2 expression and hypermethylation in this gene promoter were common genetic changes in EMPD, which expands our understanding of the role of MMR function in this skin cancer.

[Protective effects of N-acetyl-L-cysteine against binge drinking-induced fatty liver in mice].

Objective: To investigate the roles of N-acetyl-L-cysteine (NAC) against binge drinking-induced fatty liver in mice. Methods: SPF male C57BL/6 mice were randomly divided into 3 groups, i.e. control group, model group, and NAC/ethanol group (n=10). Mice in model and NAC/ethanol groups were exposed to 3 doses of ethanol (6 g/kg bw) to induced fatty liver, while mice in control group received equal volume and equal energy of maltodextrin solution. NAC was administered to mice at 1 h before ethanol exposure (100 mg/kg bw, i.p.). The mice were sacrificed at 6 h after the last ethanol exposure. The liver and epididymal adipose tissues were collected. Histopathological examination and biochemical assay kit were used to evaluate the fat accumulation, while Western-blot was performed to detect the protein levels of some key factors involved in fat metabolism in liver and adipose tissues. Results: Compored with control group mice, the liver index and liver weight were significantly increased compared with model group, the liver index and TG level in NAC/ethanol group mice were all significantly decreased (P<0.05). Histological examination showed NAC effectively suppressed binge drinking-induced fat accumulation in mice liver. In addition, NAC had no significant effects on the protein levels of peroxisome proliferator-activated receptor-α (PPAR-α), Acy-CoA oxidase (ACOX), sterol regulatory element binding protein 1 c (SREBP-1c) and fatty acid synthase (FAS). Furthermore, the protein levels of hormone sensitive lipase (HSL) did not significantly differ among 3 groups, whereas NAC prevented binge drinking-induced increase of HSL phosphorylation at ser563 and ser660. Conclusion: NAC could effectively attenuate binge drinking-induced fatty liver, which might be associated with the inhibition of lipid mobilization by suppressing the phosphorylation of HSL.

[Intravenous sedation with midazolam and propofol target controlled infusion on patients'perioperative anxiety under the mandibular third molar extraction].

OBJECTIVE: To evaluate whether midazolam with propofol target controlled infusion (TCI) intravenous sedation during the mandibular third molar extraction influences patients'perioperative anxiety. METHODS: The subjects were patients who planned to undergo the mandibular third molar extraction in Peking University School and Hospital of Stomatology, whose state anxiety inventory (SAI) scores were≥38 at the initial visit. They were divided into intravenous sedation group (IVS) and local anesthesia group (LA) on the basis of the planned intravenous sedation. Each group was divided into two subgroups according to the overall SAI scores at the initial visit: IVS-I, LA-I (SAI: 38-50) and IVS-II, LA-II (SAI: 51-80). The anxiety before and after the surgery was evaluated by the SAI scores at the initial visit (T1), before surgery (T2) and 7 days after surgery (T3). The anxiety during the surgery was evaluated by the heart rate, blood pressure and visual analogue scale (VAS) scores. RESULTS: There were no significant differences on SAI at T1, T2, and T3 in the two groups (P>0.05). The heart rate, blood pressure and VAS pain scores of IVS group were significantly lower than those of LA group during the surgery (P<0.001). CONCLUSION: Intravenous sedation with midazolam and propofol TCI was effective on the patients' anxiety during the third molar extraction, which successfully made the patients more comfortable and their heart rate, blood pressure and oxygen saturation more stable during the surgery. But there were no significant differences on the patients'anxiety at the initial visit (T1), before surgery (T2) and 7 days after surgery (T3) according to the SAI scores in the two groups.

[Mutation analysis of GJB2 gene in 1 822 patients with nonsyndromic hearing loss in Zhejiang Province].

Objective:To analyze the genetic characteristics in nonsyndromic hearing impairment (NSHL) patients in Zhejiang province.Method:Peripheral blood samples were obtained from 1822 NSHL patients and 467 normal hearing controls in Zhejiang province. We carried out a systematic mutational screening of GJB2 gene in these subjects by amplifying the coding region of GJB2 gene and sequencing directly.Result:Thirty kinds of mutation were identified, including eleven pathogenic mutations, one hypomorphic allele, sixteen polymorphic mutations and two novel mutations. The c.235delC mutation was the most prevalent pathogenic mutation in this cohort (18.50%), and the rate of allele mutation was 12.16%. The frequency of c.299_300delAT,c.176_191del16,c.512_513insAACG,c.35delG,c.283G>A,c.427C>T,c.35insG,c.439G>A,c.571T>C,c.139G>T mutations were decreased in turn.Conclusion:c.235delC mutation is the hot spot of GJB2 gene mutation in NSHL patients in Zhejiang province and the most common mutational pattern is frame-shift mutation. The discovery of novel mutations enriches the spectrum and frequency of variants in GJB2 gene.

Molecular analysis of MLH1 variants in Chinese sporadic colorectal cancer patients.

Single nucleotide polymorphisms (SNPs) in mismatch repair genes, especially in the MLH1 gene, are closely associated with susceptibility to hereditary nonpolyposis colorectal cancer. However, few relevant findings are available regarding the association between sporadic colorectal cancer (SCRC) and SNPs of MLH1 in Chinese patients. Therefore, the present study aimed to describe the pathogenic association between three important MLH1 polymorphisms and SCRC in the Chinese population. Peripheral blood samples from 156 SCRC patients and 311 healthy controls were collected. DNA was purified from peripheral blood, and the V384D, R217C, and I219V polymorphisms were evaluated using high-resolution melting analysis and direct sequencing. The association between the three important MLH1 polymorphisms and clinical pathological features of the SCRC patients was analyzed. In addition, PMS2-MLH1 protein interactions were determined by co-immunoprecipitation (Co-IP) to determine the protein functional alteration induced by these SNPs. Among the three polymorphisms, V384D was significantly associated with the risk of SCRC (OR = 31.36, P < 0.0001). The allele frequencies were 4.81 and 0.16% in the SCRC group. No association was found between SCRC and R217C, or between SCRC and I219V. Moreover, the allele frequency of R217C was significantly higher in the SCRC patients younger than 60 years than in those older than 60 years. Co-IP showed that the MLH1 R217C, V384D, and I219V variants had relative binding abilities with PMS2 of 0.59, 0.70, and 0.80, respectively, compared with the wild-type. These findings suggest that MLH1 V384D could be a promising genetic marker for susceptibility to SCRC.

Effects of genetic correction on the differentiation of hair cell-like cells from iPSCs with MYO15A mutation.

Deafness or hearing loss is a major issue in human health. Inner ear hair cells are the main sensory receptors responsible for hearing. Defects in hair cells are one of the major causes of deafness. A combination of induced pluripotent stem cell (iPSC) technology with genome-editing technology may provide an attractive cell-based strategy to regenerate hair cells and treat hereditary deafness in humans. Here, we report the generation of iPSCs from members of a Chinese family carrying MYO15A c.4642G>A and c.8374G>A mutations and the induction of hair cell-like cells from those iPSCs. The compound heterozygous MYO15A mutations resulted in abnormal morphology and dysfunction of the derived hair cell-like cells. We used a CRISPR/Cas9 approach to genetically correct the MYO15A mutation in the iPSCs and rescued the morphology and function of the derived hair cell-like cells. Our data demonstrate the feasibility of generating inner ear hair cells from human iPSCs and the functional rescue of gene mutation-based deafness by using genetic correction.

The regulatory effects of autophagy to the CNE2 cells radio-sensitization.

To investigate the effects of autophagy activator and autophagy inhibitor on the CNE2 radiation sensitivity of nasopharyngeal carcinoma cells. RNA interference technology was used to silence the atg5 gene and autophagy inhibition cell model was constructed. Rapamycin and chloroquine were treated respectively on cells with X-ray 5Gy irradiation. Cells' growth status were observed for 8 days and control group was set. The cell viability was detected by MTT assay and colony formation assay, and the cell cycle was analyzed by flow cytometry. Compared with the control group, the survival rate, clone formation rate and the survival rate of the irradiation of the other three groups were significantly lower. (P<0.05) Most cells were detected in the G0/G1 phase in the other three groups except the control group, and cells of the other two periods were less than those in the G0/G1 phase. The autophagy inhibitor or activator and atg5 silencing can be increased by CNE2 radiation therapy, however, the sensitization effect increase of autophagy activator is better than others.

[Spectrum of GJB6 variants in 318 pedigrees with non-syndromic hearing loss:one deafness pedigree carrying both GJB6 and GJB2 deletion variant].

Objective:To investigate the mutation characteristics of GJB6 (gap juction bata 6) gene in 318 Han Chinese pedigrees with non-syndromic hearing loss.Method:Polymerase chain reaction was used to detect the coding region of GJB6 gene in 318 Han Chinese pedigrees with non-syndromic hearing loss.Gene arrays and second generation sequencing were used to detect 118 genes which had reported to be accosiated with deafness in members of pedigree which possibly carried pathogenic GJB6 gene mutation.Result:Here,we have screened the mutations of GJB6 gene in 318 Han Chinese pedigrees with non-syndromic hearing loss and found one pedigree carrying both GJB6 and GJB2 gene deletion.Clinical and molecular genetic evaluation revealed the variable phenotype of hearing impairments including age-at-onset,audiometric configuration and severity in these subjects.Mutational analysis of the GJB2 and GJB6 gene coding region showed a heterozygous 235 del C of GJB2 gene and a novel 228 del G of GJB6 gene.Conclusion:GJB6 gene 228 del G variant,which occurs at a highly evolutionarily conserved nucleotide,forward the stop codon to 81 position and result in the corresponding polypeptide 181 amino acids shorter than wildtype polypeptide.In addition,GJB6 gene 228 del G absent varies among 94 unrelated Chinese controls.Our finding suggest that GJB6 gene 228 del G maybe a novel pathogenic mutation associated with non-syndromic hearing loss.

Bone turnover and articular cartilage differences localized to subchondral cysts in knees with advanced osteoarthritis.

OBJECTIVE: To investigate changes in bone structure, turnover, and articular cartilage localized in subchondral bone cyst (SBC) regions associated with knee osteoarthritis (OA). METHODS: Tibial plateaus (n = 97) were collected from knee OA patients during total knee arthroplasty (TKA). SBCs were identified using micro-computed tomography, and the specimens were divided into non-cyst (n = 25) and bone cyst (n = 72) groups. Microstructure of subchondral bone was assessed using bone volume fraction (BV/TV), trabecular number (Tb.N), structure model index (SMI) and bone mineral density (BMD). In bone cyst group, the cyst subregion, which contained at least one cyst, and the peri-cyst subregion, which contained no cysts, were further selected for microstructure analysis. Articular cartilage damage was estimated using the Osteoarthritis Research Society International (OARSI) score. The numbers of TRAP(+) osteoclasts, Osterix(+) osteoprogenitors, Osteocalcin(+) osteoblasts and expression of SOX9 were evaluated by immunohistochemistry. RESULTS: Bone cyst group presented higher BV/TV, Tb.N and SMI at subchondral bone than non-cyst group. Furthermore, cyst subregion displayed increased BV/TV and Tb.N but lower BMD and SMI than peri-cyst subregion. Histology revealed a higher OARSI score in bone cyst group. SBC exhibited a weak relationship with BV/TV, etc. The numbers of TRAP(+) osteoclasts, Osterix(+) osteoprogenitors, Osteocalcin(+) osteoblasts and expression of SOX9, were higher in bone cyst group. CONCLUSION: SBCs within knee OA are characterized by focally increased bone turnover, altered bone structure and more severe articular cartilage damage. The increased bone turnover possibly contributes to altered bone structure localized in SBC areas, and thus aggravates articular cartilage degeneration.

A randomised phase II trial of selumetinib vs selumetinib plus temsirolimus for soft-tissue sarcomas.

BACKGROUND: The MEK inhibitor, selumetinib, suppresses soft-tissue sarcoma (STS) cell proliferation in vitro. Mammalian target of rapamycin inhibitors possess modest activity against STS; however, resistance develops via MAPK pathway feedback activation. The combination of selumetinib and temsirolimus synergistically inhibits STS cell line growth. Therefore, a randomized phase II trial of selumetinib vs selumetinib plus temsirolimus was conducted. METHODS: Seventy-one adults with advanced STS who received ⩽ 2 prior chemotherapeutics were randomized to selumetinib 75 mg p.o. bid and allowed to crossover upon progression, or to selumetinib 50 mg p.o. bid plus temsirolimus 20 mg i.v. weekly, with primary endpoint of progression-free survival (PFS). RESULTS: There was no difference in PFS between the two arms for the overall cohort (median 1.9 vs 2.1 months); an improved median PFS was observed in the combination arm (N = 11) over single agent (N = 10) in the prespecified leiomyosarcoma stratum (median 3.7 vs 1.8 months; P = 0.01). Four-month PFS rate was 50% (95% confidence interval 0.19-0.81) with the combination vs 0% with selumetinib alone in the leiomyosarcoma cohort. Most common grade 3/4 adverse events with the combination were mucositis (29%), lymphopenia (26%), neutropenia and anaemia (20% each). CONCLUSIONS: While single-agent selumetinib has no significant activity in STS, the combination may be active for leiomyosarcomas.

An isolation technique to prevent the spread of tumor cells during radical gastrectomy for gastric carcinoma located on the anterior wall of the gastric antrum.

OBJECTIVE: To report an isolation technique for reducing the spread of tumor cells during radical gastrectomy for lesions located on the anterior wall of the gastric antrum. METHODS: The isolation technique involves using linear cutting staplers and a waterproof membrane to completely "block" and isolate the area to be resected. Blood samples from the portal vein and peritoneal wash samples were obtained immediately after laparotomy and during surgical resection. RT-PCR was used to determine levels of carcinoembryonic antigen (CEA) and cytokeratin 20 (CK20). Carbon nanoparticles were injected into the subserosa of the blocked region before resection to determine lymph flow out of the region. RESULTS: A total of 76 patients (median age, 59 years; range, 22-73 years), with tumors on the anterior wall of the gastric antrum were included (blocking group, n = 40; non-blocking group, n = 36). In the blocking group, the carbon nanoparticles did not flow beyond the blocking line. There were no significant differences between the groups in CEA or CK20 mRNA levels in portal vein blood or peritoneal wash fluid immediately after laparotomy. CEA and CK20 levels were significantly lower in portal vein blood in the blocking group during tumor resection. During a median follow-up of 30 months, the incidence of metastasis or recurrence in the blocking group was lower than the non-blocking group, although it did not reach statistical significance (17.9% vs. 25.0%, respectively). CONCLUSION: The blocking technique can reduce hematogenous and lymphatic spread of tumor cells into the systemic circulation, and may prevent metastasis or recurrence after radical gastrectomy for gastric carcinoma.

[Urinary BLCA-4 level is useful to detect upper urinary tract urothelial cell carcinoma]. / El nivel de BLCA-4 urinario es útil para la detección del carcinoma de células uroteliales de vía urinaria superior.

OBJECTIVE: Upper urinary tract urothelial cell carcinomas (UUT-UCCs) are rare but usually invasive at diagnosis. Early diagnosis of UUT-UCCs is thus warranted. UUT has the same embryological origin with bladder and BLCA-4 is a highly sensitive and specific marker for bladder cancer. We intend to investigate the viability of BLCA-4 in detecting UUT-UCCs. MATERIAL AND METHODS: Urines from 30 UUT-UCC patients, 10 ureteral polyp patients, 20 infected patients with incarcerated ureteral stones, and 30 normal controls were included. BLCA-4 antibody was produced and applied in an indirect ELISA assay. RESULTS: Urinary BLCA-4 is significantly higher in UUT-UCC group than «Polyp¼ group (P=0.0017), «Infection¼ group (P<0.0001), or « Normal¼ group (P<0.0001). The «Polyp¼ group is also higher than «Infection¼ group (P=0.015), or «Normal¼ group (P=0.0009). ROC curve revealed at cut-off of 5.5×10(-4)A, sensitivity was 93.3% and specificity was 100%. When grouped as ureteral mass vs normal, same cut-off value yielded 93.3% sensitivity and 83.3% specificity. At 2.4×10(-4)A, sensitivity was 56.7% and specificity was 97.2%. CONCLUSIONS: Urinary BLCA-4 is also highly specific in UUT-UCCs detection. For incidentally identified ureteral mass, BLCA-4 can be considered an auxiliary indicator besides biopsy.

BLCA-4 expression is related to MMP-9, VEGF, IL-1α and IL-8 in bladder cancer but not to PEDF, TNF-α or angiogenesis.

AIM: BLCA-4 is a specific nuclear matrix protein found in bladder cancer and there is a dearth of study on functional analysis upon this factor. We aimed to discover whether BLCA-4 is related to angiogenesis in bladder cancer. METHODS: Fifty-three bladder cancer samples were included for immunohistochemical staining of BLCA-4, matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF), interleukin-1α (IL-1α), IL-8, pigment epithelium-derived factor (PEDF), tumour necrosis factor-α (TNF-α) and von Willebrand factor (vWF) for microvessel density (MVD). Expressional levels were scored and grouped by clinicopathological parametres for statistical analysis for correlations. RESULTS: Positive correlations were identified between expression of BLCA-4 and IL-1α (p=0.038), IL-8 (p=0.001), VEGF (p=0.002), and MMP-9 (p=0.013). No correlation was found for PEDF (p=0.182), TNF-α (p=0.531) or MVD (p=0.932). Positive correlations were also obtained in cases of advanced grade or stage, larger, recurrent and multiple tumours. Positive correlation between BLCA-4 and MMP-9 was also found in papillary urothelial neoplasm of low malignant potential (PUNLMP). CONCLUSION: BLCA-4 may not effect pro-angiogenic pathways in bladder cancer, it can however interact with IL-1α, IL-8, VEGF and MMP-9 to enhance tumourigenesis and tumour invasiveness.

Urinary BLCA-4 is highly specific for detection of bladder cancer in Chinese Han population and is related to tumour invasiveness.

BLCA-4 is currently the most sensitive and specific urinary marker for bladder cancer. As the incidence of bladder cancer varies by ethnic and territory, we tended to evaluate the feasibility of bladder cancer detection using urinary BLCA-4 in Chinese Han nationality. Urines from 79 bladder cancer patients, 31 urinary tract infection patients and 29 normal controls were included. Tissue specimens of 53 bladder cancers, 24 pathologically normal tumour- adjacent urothelium and 15 healthy controls were involved. BLCA-4 antibody was produced and applied in an indirect ELISA assay for urine samples and immunohistochemistry study in tissue samples. Urinary BLCA-4 was significantly higher in the bladder cancer group (P=0.0001). The level was in no relation to age, gender, growth pattern, grade or stage. Discrepant to reported data, a cut-off value of 1.7×10⁻4 A was acquired here, which yields a sensitivity of 97.37% and specificity of 100%. Muscle invasiveness was related to a higher BLCA-4 level (P=0.0175). Tumour tissues were also scored higher in staining (P=0.0001), yet this was not related to invasiveness. In 41.67% of adjacent normal tissue we found positive BLCA-4 expression. Urinary BLCA-4 was also highly specific in bladder cancer detection in the Chinese, with additional indicative value in muscle invasiveness detection. A cut-off value of 1.7×10⁻4 A may be more adaptive to Chinese Han population.

High-resolution melting analysis of PCDH10 methylation levels in gastric, colorectal and pancreatic cancers.

Protocadherins are celladhesion molecules with 6 or 7 cadherin motifs in their extracellular domain and various cyotoplasmic domains. PCDH10 was characterized a novel tumor suppressive gene in and was epigenetically silenced in multiple haematologic malignancies as well as some solid tumors such as gastric cancer, nasopharyngeal carcinoma and esophageal carcinoma. High-resolution melting (HRM) analysis has been used as a novel tool for analysis of promoter methylation. In our study, we used HRM analysis to detect the methylation levels of PCDH10 gene in 100 gastric cancers, 100 colorectal cancers, 70 pancreatic cancers and equal number of adjacent normal tissues. The frequency of PCDH10 methylation in all three types of cancers was significantly higher than that in normal tissues. Consistent with previous reports, expression levels of PCDH10 were inversely correlated with methylation levels. But we didn't find significant association between PCDH10 methylation status and TNM staging in all three types of cancers. In summary, application of HRM analysis to large amount of clinical samples proves to be a fast and high-throughput way to investigate the epigenetic status of PCDH10. And this is the first study to evaluate the prevalence of PCDH10 methylation based on large amount of tumor samples, showing that epigenetic regulation of PCDH10 was associated with carcinogenesis.