Objective To investigate the roles and significances of MMP-2 and MMP-9 in diffuse proliferative lupus nephritis by repeated renal biopsy.Methods Seventeen patients diagnosed by renal biopsy as WHO typeⅣlupus nephritis were analyzed by immunohistochemistry staining for MMP-2 and MMP-9. Double staining for MMP-2 and MT1-MMP,MMP-9 and CD68 were also performed.Patients had repeated renal biopsy after followed up for 2.5 years.The relationship between expressions of gelatinases and pathological activity index and clinical data were studied.Results MMP-2 immunoreactivity was detected in normal controls and was increased in diffuse proliferative lupus nephritis.MMP-9 staining,which was almost negative in normal giomeruli,was increased much more significantly in diffuse proliferative lupus nephritis. The immunoreactivity of MMP-2 and MMP-9 was positive in MT1-MMP staining and CD68-positive macrophages, respectively.The expression of MMP-2 and MMP-9 was reduced by 70% and 62% in 10 patients whose clinical condition was partially alleviated,while the expressions in 7 patients whose clinical condition was not alleviated,were only reduced by 27% and 32%.The staining for MMP-2 and MMP-9 were correlated with activity index of lupus nephritis and proteinuria.Conclusion Up-regulation of gelatinases expression in diffuse proliferate lupus nephritis is correlated to activity index of the disease.

[ ABSTRACT] AIM:To investigate the effects of tanshinone IIA ( Tan IIA) on proliferation, apoptosis and its mo-lecular mechanism in human hepatoma HepG2 cells under hypoxic condition.METHODS:Hypoxia model was established by treatment with cobalt chloride ( CoCl2 ) .The cells were divided into normoxia control group, hypoxia control group and hypoxia combined at different concentrations of Tan IIA groups.After HepG2 cells were incubated with different concentra-tions of Tan IIA (0.5, 1.0, 2.0, 5.0 and 10.0 mg/L) for 24 h, 48 h and 72 h under hypoxic condition, the cell prolifer-ation was determined by MTT assay.After Tan IIA was added to the media at different concentrations for 24 h and 48 h, the apoptotic cells were observed by Hoechst 33258 staining.The protein levels of hypoxia-inducible factor 1 alpha (HIF-1α) , vascular endothelial growth factor ( VEGF) and wild-type P53 were detected by Western blotting after cultured with different concentrations of Tan IIA for 48 h.RESULTS:Tan IIA inhibited the proliferation of HepG2 cells in a dose-and time-dependent manner.Tan IIA induced the typical morphology of apoptotic cells and increased the apoptotic rate in a dose-and time-dependent manner after treatment with 1.0 mg/L~5.0 mg/L for 24 h and 48 h under hypoxic condition. The protein levels of HIF-1αand VEGF were weakly expressed in HepG2 cells under normoxia but up-regulated after incu-bated under hypoxia for 48 h.The protein expression of HIF-1αand VEGF were decreased with the increase in the concen-tration of Tan IIA under hypoxia.The protein expression of wild-type P53 was increased with the increase in the concentra-tions of Tan IIA under hypoxia.CONCLUSION:Tan IIA significantly inhibits the proliferation and induces the apoptosis of human hepatoma HepG2 cells under hypoxia, which may be related to the down-regulation of HIF-1αand VEGF and up-regulation of wild-type P53.

Aim To investigate the mechanism of demethylation on adenosine (ADO )and homocysteine (HCY)-induced apoptosis in human hepatoma HepG2 cells .Methods HepG 2 cells were treated with differ-ent concentrations of ADO (1.0、2.0、4.0 mol · L-1 ) alone or in combination with HCY for 6h,12h and 24h,5-aza-2-deoxycytidine (5-Aza-CdR)as a positive control.Cell viabilities were assessed by CCK8 assay. Cell apoptosis was observed by AnnexinV-FITC/PI staining.The mitochondrial membrane potentials(ΔΨ) were measured by flow cytometry.The mRNA and pro-tein expressions of caspase-3,caspase-8,caspase-9, MDM-2,p53,Cytochrome C,DNMT1,DNMT3a,DN-MT3 b were detected by RT-qPCR and Western blot re-spectively.Results ADO alone or in combination with HCY significantly reduced the viability of HepG2 cells in a dose and time-dependent manner.The apoptotic rates of HepG2 cells after combination treatment with ADO and HCY at 1 .0,2.0,4.0 mol · L-1 for 24 h were (1 8.63 ± 1.25 )%,(29.42 ±2.37 )% and (42.47 ±3.09 )%,compared with the control group (1.30 ±0.82 )%,P <0.01;and the mitochondrial membrane potentials were decreased from 674.15 ± 82.8%(black control group)to (428.38 ±54.5)%, (297.78 ±30.5)%,(74.45 ±5.73)%,P<0.01, respectively.The expressions of caspase-3,caspase-8, caspase-9,MDM-2,p53,Cytochrome C were up-regula-ted and MDM-2 were down-regulated after combination treatment of ADO and HCY.The mRNA expressions of DNMT1 ,DNMT3 a and DNMT3 b were down-regulated after combination treatment with ADO and HCY or 5-Aza-CdR alone.Conclusion Combination treatment of ADO and HCY can cause cellular methylation chan-ges.The effects of demethylation of ADO and HCY may activate p53 gene and mitochondrial pathway, which at last leads to HepG2 cell apoptosis.

italic>Ardisia crispa (Thunb.) A. DC. is a traditional Miao medicinal herb with significant therapeutic effects in the treatment of sore throat, tonsillitis, edema of nephritis and bruising and rheumatism, etc. Ardisia crispa var. amplifolia and Ardisia crispa var. dielsii are varieties of A. crispa. A. crispa var. amplifolia and A. crispa var. dielsii are controversial in terms of species evolutionary relationships and taxonomic identification. In this study, we sequenced the whole genome sequences of A. crispa var. amplifolia and A. crispa var. dielsii chloroplasts using Illumina platform, assembled, annotated and characterized them, compared the structural features and degree of variation among chloroplast genomes using bioinformatics methods, and also downloaded constructing phylogenetic trees to analyze the phylogenetic relationships of chloroplasts in Primulaceae and Myrsinaceae using whole genome sequence information. The results showed that the complete chloroplast genome sequences of A. crispa var. amplifolia and A. crispa var. dielsii were 156 749 bp and 156 748 bp in length, with 132 genes annotated, including 87 protein-coding genes; the codon preference of A/U was greater than that of G/C; The differences in the coding regions of rps15 and rpoB genes in the comparative genome analysis can be used as loci for molecular identification of the two species; the differences in the coding regions of ycf1, ycf2, rpoC1, ycf3, petD and rpl16 genes in the chloroplast genome compared with those of the same genus can be used as loci for identification of the genus. In the phylogenetic results, A. crispa var. amplifolia and A. crispa var. dielsii were clustered together with 100% support, indicating that they are closely related. In this research, we analyzed the chloroplast genome structure and phylogenetic relationships of A. crispa var. amplifolia and A. crispa var. dielsii, providing an important theoretical basis for their molecular identification, genetic variation, breeding and phylogenetic analysis.

OBJECTIVETo compare the clinical, pathological, laboratory test and follow-up data between familial and sporadic patients with distal myopathy with rimmed vacuoles (DMRV) and discuss the characteristics of this disorder in Chinese population.

METHODSThe clinical and pathological features, laboratory data and follow-up results of 33 sporadic and 4 familial cases of pathologically confirmed DMRV were summarized and compared retrospectively.

RESULTSThe patients age, onset age, or disease duration showed no significant difference between sporadic and familial cases; the onset pattern and affected muscle groups were also similar, but the sporadic cases showed more frequent dysmorphic features than the familial cases. The patients showed mild to moderate elevation of the muscle enzymes by one to three folds, and the familial patients had more significant elevation than the sporadic ones. No correlation was found between the disease duration and the level of muscle enzymes. The pathological findings were similar between the cases, and Gomori staining showed rimmed vacuoles and inclusion bodies without inflammatory cell infiltration. Follow-up results of 29 cases showed no significant difference between the two groups. The disease was slowly progressive and severely affected the quality of life of the patients, but did not produce obvious effect on the life expectancy.

CONCLUSIONThe clinical, pathological and laboratory data of Chinese DMRV patients are basically similar to those of Japanese cases. Sporadic cases tend to show more dysmorphic features than the familial ones, and occasional sporadic cases have early disease onset in early childhood.

Adult ; Asian Continental Ancestry Group ; Distal Myopathies ; classification ; genetics ; pathology ; Female ; Humans ; Inclusion Bodies ; pathology ; Male ; Pedigree ; Retrospective Studies ; Vacuoles ; pathology ; Young Adult

Objective To investigate the efficacy and white matter integrity of low frequency repetitive transcranial magnetic stimulation (rTMS) and the modified electroconvulsive therapy (MECT) in patients with schizophrenia. Methods From May 2015 to October 2016,120 cases with schizophrenia were randomly enrolled into the MECT group and of the rTMS group. Patients in the MECT group were treated with the modified electric convulsive therapy for 8 times ,while patients in the rTMS group were treated with the repetitive transcranial magnetic stimulation for 12 times. PANSS were used to evalue the clinical effects. Repeatable battery for the assess-ment of neuropsychological status was used to assess the cognitive function. Treatment emergent side-effect scale was used to assess the adverse effects. Brain fractional anisotropy was used to assess white matter integrity. Results After treatment,the PANSS scores were significantly lowered,however,the RBANS scores were signifi-cantly higher in the MECT group and rTMS group than those before treatment ,with significant differences (P <0.05). No significant differences for the PANSS scores and the RBANS scores were observed between the two groups before and after treatment. There was no significant difference for the TESS scores between the two groups before treatment(P > 0.05). After treatment,the TESS scores in the MECT group were significantly higher than those in the rTMS group(P < 0.05). After treatment,the FA values of left anterior cingulate gyrus,left posterior cingulate gyrus ,left prefrontal cortex and genu of corpus callosum for both MECT group and rTMS group were significantly increased (P < 0.05 ,respectively). Compared with the MECT group ,the FA value significantly increased in the rTMS group after treatment (P < 0.05). Conclusions Both MECT and rTMS have significant clinical efficacies and can improve the cognitive function of schizophrenics. rTMS is more safe than MECT ,with a stronger effect on preventing the integrity of white matter than MECT.

OBJECTIVETo investigate the feasibility of differentiation of human induced pluripotent stem cells (iPSCs) into epidermal-like stem cells.

METHODS(1) Human strain of iPSCs were plated on-to trophoblast of inactivated Fb strain of mouse embryos and cultured in complete medium of embryonic stem cells, iPSCs were subcultured by collagenase IV digestion method. The morphology and growth of iPSCs were observed under inverted phase contrast microscope, and the cells were stained with alkaline phosphatase (AKP). iPSCs were cultured in incomplete medium of embryonic stem cells to observe the ability of embryoid body formation. (2) Human iPSCs were inoculated onto 6-well plate covered with human amniotic membrane to culture as induction group. Other iPSCs were cultured on 6-well plate without human amniotic membrane as control group. Morphological changes in iPSCs in two groups were observed. Expressions of integrin beta1 and CK19 of iPSCs in two groups were determined by immunocytochemical staining.

RESULTSHuman iPSCs showed a typical stem cell clone-like growth with a clear boundary, and they proliferated vigorously in complete medium of embryonic stem cells. These cells were AKP-positive. iPSCs formed embryoid body in trophoblast-free and suspension culture conditions. After 4 days of co-culture, stem cell clones were formed on the surface of amniotic membrane in induction group, and part of the cells were integrin beta1 and CK19 positive. Most of the cells died, and no integrin beta1 and CK19 positive cells were found in control group.

CONCLUSIONSHuman iPSCs can be differentiated into epidermal-like stem cells by amniotic membrane induction, and it lays an experimental basis for providing new source of seed cells of skin tissue engineering.

Animals ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Epidermis ; cytology ; Humans ; Induced Pluripotent Stem Cells ; cytology ; Mice

OBJECTIVETo investigate the prognostic factors in non-small cell lung cancer (NSCLC) at stage III and IV and establish a reliable model of clinical prognostic index.

METHODSKaplan-Meier and Cox regression were used to analyze the relationship between the prognostic factors and survival time in 114 cases of NSCLC. The prognostic factors included clinical-pathological features and serum levels of cytokeratin fragment 19 (Cyfra21-1), CEA, neuron-specific enolase (NSE), CA125, interleukin-2 (IL-2) and soluble interleukin-2 receptors (sIL-2R).

RESULTSKaplan-Meier analysis showed that KPS, sex, disease stage, treatment, Cyfra21-1, sIL-2R and CA125 were related to prognosis. Multivariate analysis indicated that Cyfra21-1, stage and treatment were independent prognostic factors. When Cyfra21-1 > 3.5 mg/L, stage IV and chemotherapy < 3 cycles, the relative risk (RR) was 1.691, 2.229 and 3.035, respectively. In patients given 3 or more cycles of chemotherapy, serum Cyfra21-1, sIL-2R and stage at diagnosis were significantly independent prognostic factors. Three of these prognostic factors were used to establish a prognostic index (PI) model based on a simple algorithm: PI = Cyfra21-1 + sIL-2R + stage. The median survival period of patients with 3 or more cycles of chemotherapy were 18 months if PI = 0, 8 months if PI = 1 or 2, and 5 months if PI = 3.

CONCLUSIONThe serum Cyfra21-1, sIL-2R and disease stage in unresectable NSCLC were independent prognostic factors. PI calculated on the basis of Cyfra21-1, sIL-2R and stage is recommended to predict the survival period of NSCLC.

Antigens, Neoplasm ; blood ; Biomarkers, Tumor ; blood ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; mortality ; pathology ; Female ; Follow-Up Studies ; Humans ; Keratin-19 ; Keratins ; Lung Neoplasms ; drug therapy ; mortality ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Proportional Hazards Models ; Receptors, Interleukin-2 ; blood ; Survival Rate

OBJECTIVETo evaluate the static and dynamic contrast sensitivity changes in myopic patients before and after laser in situ keratomileusis (LASIK).

METHODSSeventy-three eyes in 37 patients with myopia (with or without astigmatism) who received LASIK were tested for static and dynamic contrast sensitivities using the METRO VISION MON ELEC I system at 0.7, 1.4, 2.7, 5.5, 11, and 22 cpd and cps prior to LASIK, and at one-, three-, and six-month intervals after LASIK.

RESULTSAll eyes gained naked visual acuity of more than 0.5 after LASIK. The contrast sensitivity was depressed at all frequencies 1 month after LASIK, as compared to one week prior to LASIK. The depression at 2.7, 5.5, 11 (P < 0.01) and 22 cpd (P < 0.05) was statistically significant for static contrast sensitivity, and also at 5.5 (P < 0.01) and 11 cps (P < 0.05) for dynamic contrast sensitivity. Myopic eyes between 6.25 D and 14.0 D, and astigmatic eyes 2 DC and more, suffered more static and dynamic contrast sensitivity depression than the myopic eyes between 1.25 D and 6.00 D and astigmatic eyes less than 2 DC. Contrast sensitivities were improved and exceeded preoperative levels 3 months after LASIK, and improved even more 6 months after LASIK. All sequences were statistically significant for static contrast sensitivity (P < 0.01), while only 2.7, 5.5, and 11 cps were statistically significant for dynamic contrast sensitivity (P < 0.01). The astigmatic eyes 2 DC and more showed less improvement, even below the preoperative level at 1.4 cps of dynamic contrast sensitivity.

CONCLUSIONSWhile temporary depression of contrast sensitivity for myopic eyes after LASIK was seen, contrast sensitivity soon returned to exceed preoperative levels at 3 months after LASIK, while improving even more 6 months after LASIK.

Adolescent ; Adult ; Astigmatism ; surgery ; Contrast Sensitivity ; Cornea ; surgery ; Female ; Humans ; Keratomileusis, Laser In Situ ; Male ; Myopia ; physiopathology ; surgery ; Visual Acuity

Objective:To observe the efficacy of the serial treatment with autologous hematopoietic stem cell transplantation after bortezomib and dexamethasone-based triple chemotherapy regimen and followed by lenalidomide and intermittent intensive therapy in primary plasma cell leukemia.Methods:A retrospective analysis was made on the clinical data of one patient who was diagnosed as primary plasma cell leukemia with complex karyotype in April 2018 in Henan Cancer Hospital, and the relevant literature was reviewed.Results:The patient received multiple cycles of bortezomib and dexamethasone-based triple chemotherapy regimen, then received autologous hematopoietic stem cell transplantation, lenalidomide and intermittent intensive therapy. The patient eventually achieved complete remission and the progression-free survival time was 18 months until the day before the deadline for this article.Conclusion:The treatment with autologous hematopoietic stem cell transplantation after bortezomib and dexamethasone-based triple chemotherapy regimen and followed by lenalidomide and intermittent intensive therapy may improve the prognosis of patients with primary plasma cell leukemia and prolong the survival time.