Characteristics and measurement principles of reference methods in clinical biochemistry were described.Implementation of reference systems is one of the most effective approaches to improve the accuracy and comparability of clinical laboratory test results.Reference methods are the key components of reference systems.Reference methods should have measurement uncertainties that meet the requirements of the intended use,and thus should be based on reliable measurement principles.For the well-defined biochemistry analytes,reference methods have been almost all based on instrumental analysis.Isotope dilution mass spectrometry (ID/MS) is considered most reliable and has been the major analytical principle of the reference methods.ID/MS analysis is accurate but expensive.Use of other validated instrumental analyses as reference measurement principles would be justified.

0.05).Conclusions The ?2AR Gly16Gly genetic polymorphism was correlated with asthma severity and may be one of susceptibility genes in severe asthmatic children in Shanghai area.

OBJECTIVETo study the inhibitory effect and mechanism of Ganoderma lipsiense extract (GLE) on the growth of triple-negative breast cancer (TNBC) cell line MDA-MB-231-HM in a mouse model.

METHODSThe mouse model of TNBC was established by subcutaneous injection of 1.5 x 10(6) of MDA-MB-231-HM cells into BALB/c-nu mouse. Twenty successfully modeled mice were divided into the GLE group and the negative control group according to random digit table, 10 in each group. GLE (0.2 mL 100 mg/mL) was peritoneally injected to mice in the GLE group, while equal dose of normal saline was peritoneally injected to mice in the negative control group. The medication was administered once per 3 days and discontinued after 45 days. The CD34 expression was detected using immunohistochemical assay for counting microvessels. Meanwhile, expressions of thrombospondin 1 (TSP-1) and cyclin D1 were detected using immunohistochemical assay.

RESULTSThe average weight was obviously lower in the GLE group than in the negative control group [(0.33 ± 0.16) g vs (0.68 ± 0.37)g, P < 0.05]. The tumor inhibition rate was 51.4% in the GLE group. The volume of transplanted tumor was obviously lesser in the GLE group than in the negative control group (P < 0.05). Results of immunohistochemical staining showed, the microvessel density (MVD) under every field was (20.7 ± 2.1), TSP-1 positive cell count was (66.2 ± 9.2), cyclin D1 positive cell count was (33.8 ± 16.4) in the GLE group, and they were 34.0 ± 2.0, 24.0 ± 6.6, and 168.2 ± 32.6, respectively in the negative control group. There was statistical difference in all indices between the two groups (P < 0.05).

CONCLUSIONGLE could inhibit malignant proliferation of tumor cells by suppressing angiogenesis of blood vessels in tumor tissues and regulating cell cycles, thereby inhibiting TNBC.

Animals ; Biological Products ; pharmacology ; Cell Line, Tumor ; Cyclin D1 ; metabolism ; Disease Models, Animal ; Ganoderma ; chemistry ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microvessels ; Neoplasm Transplantation ; Neovascularization, Pathologic ; prevention & control ; Random Allocation ; Thrombospondin 1 ; metabolism ; Triple Negative Breast Neoplasms ; drug therapy

OBJECTIVETo study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for repairing articular cartilage injury in vivo.

METHODSRabbit bone marrow mesenchymal stem cells (BMSCs) were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treatment. After animals anesthesia,a full-thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile, the transfected cells were implanted to repair the rabbit model with full-thickness cartilage defects. Cartilage defects tissue was observed with light microscope, electron microscope, HE and immunohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation.

RESULTSAt 3 days after the transfection, Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection, the expression of collagen type II began and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox9 gene. Histological observation showed that at 6 weeks after the operation, the defects in the experimental group was filled with hyaline like cartilage tissue, 12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively, the defects were filled with fibrous tissues in control groups. Meanwhile, immunohistochemical staining of sections with type II collagen antibodies showed the proteins in the regenerated tissue stained positive for type II collagen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances.

CONCLUSIONOverexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells (BMSCs) promote the repair of cartilage defect.

Animals ; Bone Marrow Cells ; metabolism ; Bone Marrow Transplantation ; Cartilage, Articular ; injuries ; metabolism ; Cell- and Tissue-Based Therapy ; Female ; Genetic Vectors ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Osteoarthritis ; genetics ; metabolism ; therapy ; Rabbits ; SOX9 Transcription Factor ; genetics ; metabolism ; Tissue Engineering

AIM:To evaluate the optic nerve and axon impairment of relapsing - remitting multiple sclerosis ( RRMS) and neuromyelitis optica spectrum disorders ( NMOSD ) via detecting the peripapillary retinal nerve fiber layer (pRNFL) and the ganglion cell complex( GCC) thickness by optic coherence tomography(OCT).
METHODS: Retrospective case control study. Two hundred three cases were collected from August 2014 to January 2016 in Beijing Tian Tan Hospital. They were divided into four groups, including the normal group (n=60), the RRMS group ( n = 60 ), the NMOSD anti -aquaporin- 4 autoantibody seropositive( NMOSD- AQP4 -Ab seropositive) group (n= 48), and the NMOSD-AQP4-Abseronegative group (n = 35). All people were detected for the average and four quadrants ( superior, inferior, nasal, temporal) of pRNFL thickness and the average and two quadrants (superior, inferior) of GCC thickness with OCT. One way analysis of variance or nonparametric tests was used to compare the differences of pRNFL and GCC thickness between groups.
RESULTS: Comparing with the normal group, the average and all quadrants of pRNFL and GCC thickness in the RRMS, the NMOSD - AQP4 - Ab seropositive and the NMOSD-AQP4-Ab seronegative group were thinner (P<0. 01). Among them, the pRNFL and GCC thickness in the NMOSD- AQP4 - Ab seropositive group was the thinnest. Differences between groups in the pRNFL thickness:compared with the RRMS group, all quadrants of pRNFL and GCC thickness in the NMOSD-AQP4-Ab seropositive group were significantly thinner(P<0. 01); compared with the NMOSD- AQP4- Ab seronegative group, the inferior, nasal and temporal pRNFL thickness in the NMOSD-AQP4-Ab seropositive group were significantly thinner(P<0. 05), while the superior quadrant did not show significant differences( P > 0. 05); compared with the RRMS group, the superior pRNFL thickness in the NMOSD - AQP4 - Ab seronegative group was significantly thinner ( P < 0. 05), while the inferior, nasal and temporal quadrants did not show significant differences ( P > 0. 05 ). Differences between groups in the GCC thickness: compared with both the RRMS and the NMOSD- AQP4- Ab seronegative group, all quadrants of GCC thickness in the NMOSD -AQP4-Ab seropositive group were significantly thinner (P<0. 05); compared with the RRMS group, the superior GCC thickness in the NMOSD - AQP4 - Ab seronegative group was significantly thinner(P<0. 01), while the inferior quadrant did not show significant difference(P>0.05).
CONCLUSION: The optic nerve and axon impairment in NMOSD - AQP4 - Ab seropositive group was the most severe and the impairment in RRMS group was the least severe. The impairment in NMOSD - AQP4 - Ab seronegative group was between the former two, and could be more similar to that of RMMS.

Adolescent ; Adult ; Aged ; Cerebellar Neoplasms ; physiopathology ; Cerebellopontine Angle ; Child ; Humans ; Male ; Middle Aged ; Vestibular Evoked Myogenic Potentials ; Young Adult

Cloning, Molecular ; Escherichia coli ; genetics ; Hepacivirus ; genetics ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Viral Nonstructural Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVETo evaluate the correlation between programmed death-ligand 1 (PD-L1) expression in primary lung cancer cells, tumor associated macrophages (TAM) and patients' clinicopathological characteristics.

METHODSFrom 2008 to 2010, 208 non-small cell lung cancer patients who underwent surgery or CT-guided biopsy were recruited from Huadong Hospital, Fudan University. Immunohistochemistry staining was performed to evaluate the PD-L1 expression in both primary lung cancer cells and CD68 positive TAM. The relationship between PD-L1 expression and the clinical pathology was evaluated using χ(2) test. Spearman's rank correlations were used to determine the correlation between PD-L1 expression in tumor cells and macrophages.

RESULTSPositive PD-L1 expression in primary cancer cells was found in 136 (65.3%) patients, which were negatively correlated with lymph node metastasis (P=0.009) and smoking history (P=0.036). Besides, TAM with PD-L1 expression (found in 116 patients) was positively associated with smoking history (P=0.034), well-differentiation (P=0.029) and negative lymph node metastasis (P=0.0096). A correlation between PD-L1 expression in primary tumor cells and non-small cell lung cancer associated macrophages was found (r=0.228, P=0.021).

CONCLUSIONPD-L1, secreted from TAM, might induce cancer cells apoptosis, and decrease lymph node metastasis.

Adult ; Aged ; Aged, 80 and over ; Apoptosis ; B7-H1 Antigen ; secretion ; Carcinoma, Non-Small-Cell Lung ; pathology ; secretion ; Cell Line, Tumor ; Female ; Humans ; Lung Neoplasms ; pathology ; secretion ; Lymphatic Metastasis ; Macrophages ; pathology ; secretion ; Male ; Middle Aged ; Retrospective Studies

Objective To observe the clinical efficacy of arthroscopy and fibula osteotomy in the treatment of medial knee osteoarthritis.Methods Thirty-six patients with medial knee osteoarthritis treated in our hospital from September 2014 to September 2016 were divided into two groups according to the acceptable surgical protocol.The control group was treated by fibula osteotomy,and the observation group received arthroscopy combined with fibula osteotomy.The clinical signs and symptoms,VAS and ISOA scores were evaluated and compared between the two groups.Results The control group of 18 cases,clinical recovery in 0 cases,markedly effective in 7 cases,effective in 8 cases,3 cases with no effect,the effective rate was 38.9％;observation group of 18 cases,3 cases clinically cured,effective in 10 cases,effective in 5 cases,the effective rate was 72.2％.The effective rate of observation group was better than that of the control group,the difference was signicant (P ＜ 0.05).VAS and ISOA in both groups were significantly improved after operation,the difference was signicant (P ＜ 0.05);VAS improvement in observation group was better than that in control group,the difference was signicant(P ＜ 0.05),ISOA score in observation group was significantly better than that in control group,the difference was signicant(P ＜0.01).The clinical signs and symptoms(pain and numbness,walking,living and working abilities,tenderness,straight leg raising test,knee and Achilles reflex,total score) in two groups were improved respectively compared with those before treatment,and the differences were statistically significant(P ＜ 0.05).The pain and numbness,walking,tenderness,straight leg raising test and total score in the observation group were significantly different from those in the control group(P ＜ 0.05).Conclusion The arthroscopy and fibula osteotomy is safe and feasible to treat patients of medial knee osteoarthritis with less traumatic,more effective and better functional recovery.

Objective To analyze the genetic diversity of Pomacea canaliculata based on the mitochondria DNA cytochrome c oxidase subunitⅠ(mtDNA COⅠ)gene as a molecular marker in Lincang City of Yunnan Province,so as to provide the scien-tific data for monitoring Angiostrongylus cantonensis in local areas. Methods The genotypes and polymorphisms of 38 speci-mens of P.canaliculata collected from Mengding Town of Lincang City were analyzed by sequencing COⅠgene.The phylogenet-ic tree and genetic distances were produced based on the haplotypes from GenBank and the present study by using the neighbour-joining method with the software MEGA version 6.06. Results Totally 31 sequences were acquired in the present study and they produced 3 unique haplotypes.Haplotype 1 showed a higher frequency compared to the others and it accounted for 83.9 % (26/31).The data showed that the least genetic distances ranged from 0 to 0.052 between P.canaliculata and 3 haplotypes,as well as the largest genetic distances ranged from 0.021 to 0.239 between Pila conica and 3 haplotypes.Otherwise,the analysis of the phylogenetic trees based on COⅠgene sequences of P.canaliculata indicated that all of 3 haplotypes clustered into one big clade with that from Japan(GenBank accession number: AB433769),China(GenBank accession number: KT313034)and USA(GenBank accession number:EU523129),which owned the closet relationship amongst them.Their genetic relationships were distantly related to the GenBank's reference sequences of P.insularum(GenBank accession number:EF514942),P.cam-ena(GenBank accession number: EF515059)and so on. Conclusion There is a P. canaliculata species in Lincang City of Yunnan Province as well as a high genetic diversity amongst the acquired 3 haplotypes in this study.