The widespread application of next generation sequencing (NGS) in clinical settings has enabled testing, diagnosis, treatment and prevention of genetic diseases. However, many issues have arisen in the meanwhile. One of the most pressing issues is the lack of standards for reporting genetic test results across different service providers. The First Forum on Standards and Specifications for Clinical Genetic Testing was held to address the issue in Shenzhen, China, on October 28, 2017. Participants, including geneticists, clinicians, and representatives of genetic testing service providers, discussed problems of clinical genetic testing services across in China and shared opinions on principles, challenges, and standards for reporting clinical genetic test results. Here we summarize expert opinions presented at the seminar and report the consensus, which will serve as a basis for the development of standards and guidelines for reporting of clinical genetic testing results, in order to promote the standardization and regulation of genetic testing services in China.

To investigate the metabolism of quercitrin in rat intestinal flora and possible biological pathways, laying the foundation for the metabolic mechanism of traditional Chinese medicine glycosides ingredients. UPLC-Q-TOF-MS/MS method was established to detect the quercitrin and its metabolites with 0.1% formic acid solution(A)-0.1% formic acid acetonitrile(B) as the mobile phase for gradient elution at a flow rate of 0.3 mL•min⁻¹. Electrospray negative ion mode was applied to analyze the metabolites of quercitrin in rat intestinal flora. Metabolite ToolsTM, mass defect filter(MDF) and other technologies were used to screen, analyze the metabolites and infer the chemical formula of the metabolites. The results showed that quercitrin would have degalactoside, deoxygenation and acetylation reactions, and the aglycone quercetin resulted from degalactoside would have further reactions such as hydroxylation, deoxygenation, reduction, and ring opening to achieve deoxygenation metabolite kaempferol, C2-C3 double bonds hydrogenation and reduction product taxifolin, and degalactoside product quercetin. The research results showed that quercitrin can be metabolized by rat intestinal flora, which could increase their hydrophobicity and chemical diversity.

OBJECTIVETo investigate the efficacy and clinical safety of posaconazoleon primary antifungal prophylaxis against invasive fungal disease (IFD) in patients with stem cell transplantation.

METHODSAt the start from preconditioning regimen, 45 patients without IFD were administered with posaconazoleon until neutrophils greater than 0.5×10/L, 35 patients treated with micafungin were enrolled in control group. The incidence, risk factors of IFD and side effects of medicines were evaluated.

RESULTSOf the total 80 patients, 13(16%) had IFD within 100 days after allo-HSCT. The overall survival was significantly different between patients with or without IFD by Kaplan-Meier survival curve analysis (P<0.05). Out of the 45 cases in posaconazoleon group, IFD occurred in 4 cases (9%). In contrast, the incidence of IFD in control group was 26%(9 out of 35) (P<0.05). The risk factors of IFD and side effects were not significantly different between 2 groups(P>0.05).

CONCLUSIONThe primary prevention efficancy of IFD by posaconazoleon after allo-HSCT is much better than that of micafungin with well tolerability and satisfactory efficacy.

OBJECTIVETo analyze the effects of salvianolate on myocardial infarction in a murine in vivo model of ischemia and reperfusion (I/R) injury.

METHODSMyocardial I/R injury model was constructed in mice by 30 min of coronary occlusion followed by 24 h of reperfusion and pretreated with salvianolate 30 min before I/R (SAL group). The SAL group was compared with SHAM (no I/R and no salvianolate), I/R (no salvianolate), and ischemia preconditioning (IPC) groups. Furthermore, an ERK1/2 inhibitor PD98059 (1 mg/kg), and a phosphatidylinositol-3-kinase (PI3-K) inhibitor, LY294002 (7.5 mg/kg), were administered intraperitoneal injection (i.p) for 30 min prior to salvianolate, followed by I/R surgery in LY and PD groups. By using a double staining method, the ratio of the infarct size (IS) to left ventricle (LV) and of risk region (RR) to LV were compared among the groups. Correlations between IS and RR were analyzed. Western-blot was used to detect the extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (AKT) phosphorylation changes.

RESULTSThere were no significant differences between RR to LV ratio among the SHAM, I/R, IPC and SAL groups (P>0.05). The SAL and IPC groups had IS of 26.1%±1.4% and 22.3%±2.9% of RR, respectively, both of which were significantly smaller than the I/R group (38.5%±2.9% of RR, P<0.05, P<0.01, respectively). Moreover, the phosphorylation of ERK1/2 was increased in SAL group (P<0.05), while AKT had no significant change. LY294002 further reduced IS, whereas the protective role of salvianolate could be attenuated by PD98059, which increased the IS. Additionally, the IS was not linearly related to the RR (r=0.23, 0.45, 0.62, 0.17, and 0.52 in the SHAM, I/R, SAL, LY and PD groups, respectively).

CONCLUSIONSalvianolate could reduce myocardial I/R injury in mice in vivo, which involves an ERK1/2 pathway, but not a PI3-K signaling pathway.

Animals ; Blotting, Western ; Cardiotonic Agents ; pharmacology ; therapeutic use ; Flavonoids ; pharmacology ; Heart Ventricles ; drug effects ; pathology ; MAP Kinase Signaling System ; drug effects ; Male ; Mice, Inbred C57BL ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Myocardial Reperfusion Injury ; drug therapy ; enzymology ; pathology ; Organ Size ; drug effects ; Phosphorylation ; drug effects ; Plant Extracts ; chemistry ; pharmacology ; therapeutic use ; Protein Kinase Inhibitors ; pharmacology ; Staining and Labeling

OBJECTIVETo explore the mechanism of ABO discrepancy in a patient by ABO genotyping and the reasonable blood transfusion strategy.

METHODSRoutine serological test was carried out to analyze ABO blood group. The presence of the blood group determinants on the red blood cells were determined by adsorption-elution test. Exons 1-7 and adjacent introns of the ABO gene were amplified by PCR and sequenced.

RESULTSThe patient showed ABO forward and reverse typing discrepancy. ABO forward typing defined as B, however, the reverse typing indicated that the patient was AB subtype. Absorption-elution test confirmed weak A antigens on the patient's red blood cells. The ABO gene sequencing showed a T>C exchange at position in exon 7 which resulted in a isoleucine to threonine substitution at codon 256. The ABO blood group genotype was ABO*Ael05/B101.

CONCLUSIONThe 767 T>C substitution in the gene of α-1,3-N-acetyl galactose is the molecular mechanism leading to the decrease expression of A antigen of the Ael05 subtype.

OBJECTIVETo investigate the distribution of T helper (Th9) cells and its relationship with clinical characteristics of patients with acute myeloid leukemia (AML), and to analyze the activating levels of different transcriptional factors in Th9 cells.

METHODSThe peripheral blood specimens of 102 AML patients and 83 healthy persons as controls were collected, then the T cells of peripheral blood in AML patients and controls were isolated by using CD3 magnetic beads, the mRNA expression of IL-9 was detected by real-time quantitative PCR, the Th9(CD4IL-9) cell levels in diffrent stages and activating level of Th9 coexpression with IL-9 were detected by flow cytometry.

RESULTSThe mRNA expression of IL-9 in peripheral blood of AML M2 and M3 patients was significantly higher than that in control groups (P<0.01), at same time the CD4IL-9cell rate was significantly higher than that in control group also(P<0.01). The results of dynamically monitoring the distribution of Th9 cells in AML-M2 and M3 patients showed that the Th9 cell rate and the mRNA expression of IL-9 in newly diagnosed M2 and M3, and relapsed M2 groups were significantly higher than those of M2 and M3 in remission (P<0.01); the detection results of IL-9co-expression with transcriptional factors (SMAD3, IRF-1and IRF-4) indicated that the percantage of Th9 pSMAD3cells in peripheral blood of newly diagnosed and relapsed M2 and M3 patients was significantly higher than that in M2 and M3 patients in remission (P<0.01); on the contrary, the percentage of Th9 IRF-1cells in peripheral blood of M2 and M3 patients in remission was significantly higher than that in newly diagnosed M2 and M3 patients (P<0.01).

CONCLUSIONThe distribution of T helper cells in peripheral blood of AML-M2 and M3 patients significantly increases, moreover, correlates with disease status. The prediction of Th9 cell functions should be performed in combination with it transcriptional factors which have inmportant significance for microenvironment of tumors in AML patients.

OBJECTIVETo explore the expression of CD27 antigen in patients with multiple myeloma(MM), and its clinical diagnostic value, as well as the correlation of CD27 with clinical features and genetic abnormalities.

METHODSUsing 8 color flow cytometry, the immunophenotype of 123 MM patients' marrow cells was examined, while 51 cases of normal plasma cell reactive prolieration were used as control. Antibodies are as follows:ckappa-FITC/clambda-PE/ CD38-ECD/CD45-PERCP/CD19-PC7/ CD27-APC/CD138-BV421/ CD56-BV510. For differentiation of abnormal plasma cells from normal or reactive plasma cells, FS INT /FS PEAK with CD138/CD38 gating strategy was used to measure cell markers CD138,CD38, CD56, CD19, ckappa, clambda and CD45, then the expression rate of CD27 and mean fluorescence intensity(MFI) were analyzed for its diagnostic value in MM. At the same time, the laboratory data of 107 cases of MM patients were analyzed and the fluorescence in situ hybridization (FISH) was used to detecte 1q21 amplification, 13q14 deletion, p53 deletion and IGH rearrangement in 34 cases, then the clinical features and genetic abnormalities were compared between CD27and CD27MM patients.

RESULTSThe CD27 positive rate of abnormal plasma cells in MM patients was 48% (59/123) (percentage ≥20% as positive criterion), MFI was 31.3±35.6; while the positive rate of normal or reactive plasma cells were 100% (51/51), the MFI was 93.7±6.3. The positive rate and MFI of CD27 in MM patients were significantly lower than that in normal or reactive plasma cells (P<0.01). Laboratory examination of 58 cases of CD27 negative and 49 cases of CD27 positive MM patients indicated that no significant differences were shown on disease progress parameter, such as hemoglobin, albumin, serum calcium, serum creatinine,and no notable differences were involved in the analysis of prognostic factors between the 2 groups, such as β2-MG microglobulin and LDH levels (P<0.05). The 1q21 amplification, 13q14 deletion, p53 deletion, and IGH rearrangement results all were not significantly different between 17 cases of CD27 negative and 17 cases of CD27 positive MM patients(P<0.05).

CONCLUSIONCD27 has a unique expression profile, and its negative or weak expression is highly suggestive for MM. The 8 color flow cytometry can be used to analyze the expression of multiple antigens, which can provide a reliable evidence for the diagnosis and differential diagnosis of MM disease.

OBJECTIVEThe aim of this study was to investigate the effects of SiO2 on fibrocytes and whether fibrocytes participate in silicosis in vivo.

METHODSA macrophagocyte (AM)/fibrocyte coculture system was established, and AMs were treated with 100 μg/mL SiO2. Flow cytometry was used to detect the number of fibrocytes. Real-time PCR was performed to measure the expression of collagen I, collagen III, and α-SMA mRNA. The levels of collagen I, collagen III, and TGF-β1 protein were determined by ELISA. Immunohistochemical staining was performed to measure α-SMA protein expression. A rat silicosis model was induced by intratracheal instillation of SiO2. Lung histopathological evaluation was conducted using HE and Masson's trichrome staining after 1 and 9 weeks. The number of fibrocytes in peripheral blood or lung tissue of rat was detected by flow cytometry. Double-color immunofluorescence was applied to identify fibrocytes in the lung tissue.

RESULTSPeripheral blood monocytes were found to differentiate into fibrocytes in vitro in a time-dependent manner, and exposure to crystalline silica might potentiate fibrocyte differentiation. In addition, fibrocytes were able to migrate from peripheral blood to the lung tissue, and the number of fibrocytes was increased after SiO2 exposure.

CONCLUSIONSilica exposure potentiates fibrocyte differentiation, and fibrocytes may participate in silicosis in vivo.

Animals ; Cell Differentiation ; drug effects ; Collagen ; metabolism ; Fibroblasts ; drug effects ; Lung ; metabolism ; pathology ; Male ; Rats ; Silicon Dioxide ; toxicity ; Silicosis ; metabolism ; pathology

OBJECTIVETo evaluate whether mono (2-ethylhexyl) phthalate (MEHP) affects genomic DNA methylation and the methylation status of some specific genes such as patched gene (PTCH) and smoothened gene (SMO) in LNCaP cells.

METHODSLNCaP cells were treated with MEHP (0, 1, 5, 10, and 25 μmol/L) for 3 days. An ELISA assay was preformed to detect genomic methylation, including 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) content. A pyrosequencing assay was applied to assess DNA methylation in PTCH and SMO gene promoters. The correlation between DNA methylation and gene expression was assessed.

RESULTSThe proportion of cytosines with 5-mC methylation in LNCaP cells was significantly decreased by MEHP (1, 5, 10, and 25 μmol/L) in a dose-dependent manner (P < 0.01). For genes in the Hedgehog pathway, there was no significant MEHP concentration-dependent difference in the DNA methylation of PTCH and SMO.

CONCLUSIONMEHP might affect the progression of prostate cancer through its effect on global DNA methylation.

Antineoplastic Agents ; chemistry ; pharmacology ; Cell Line, Tumor ; DNA Methylation ; Humans ; Male ; Phthalic Acids ; chemistry ; Prostatic Neoplasms ; metabolism

OBJECTIVETo investigate the effect of joint mobilization on postoperative wrist joint function, pain and grip strength for elderly patients with distal radius fracture.

METHODSFrom January 2015 to June 2016, a total of 67 elderly patients with distal radius fracture were randomly divided into routine exercise group and joint mobilization group. Among them, 37 patients in the routine exercise group underwent conventional distal radius fracture postoperative joint function exercise regimen, including 16 males and 21 females with a mean age of (67.8±3.2) years old ranging from 60 to 72 years old;the injured side was dominant in 23 cases and non-dominant in 14 cases;injury mechanism was fall in 26 cases, traffic accident in 11 cases; for AO type, 6 cases were type B3, 18 cases were type C1, 7 cases were type C2, 6 cases was type C3. Other 30 patients in the joint mobilization group underwent joint mobilization on the basis of the routine exercise group including 14 males and 16 females with a mean age of (67.1±4.0) years old ranging from 61 to 74 years old; the injured side was dominant in 21 cases and non-dominant in 9 cases;injury mechanism was fall in 25 cases, traffic accident in 5 cases;for AO type, 8 cases were type B3, 13 cases were type C1, 6 cases were type C2, 9 cases were type C3. The wrist joint activity, Gartland-Werley wrist joint function score, VAS pain score and grip strength were observed at 3 months afrer treatment.

RESULTSAfter 3 months' treatment, the VAS in the routine exercise group was higher than that of the joint mobilization group (<0.05). The grip strength of affected side in both groups were lower than that of contralateral side, but the average grip strength of affected side in joint mobilization group was higher than that in routine exercise group(<0.05). In routine exercise group, the average angle of flexion, extension, radial deviation were significantly higher than those of joint mobilization group(<0.05). But ulnar deviation angle in routine exercise group compared with joint mobilization group had no significant difference (>0.05). In the comparison of each item of Gartland-Werley, there was no significant difference between two groups in residual deformity and complication(>0.05); the average score of subjective score, objective score and total score in routine exercise group were significantly higher than those of the joint mobilization group (<0.05). The wrist function Gartland-Werley score in routine exercise group after treatment was excellent in 21 cases, good in 10, 6 in fair, while in joint mobilization group, excellent in 23, good in 6, fair in 1(<0.05).

CONCLUSIONSThe application of joint mobilization in the treatment of elderly patients with distal radius fracture can improve the joint activity and obtain better wrist function after surgery.