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Artificial intelligence-assisted blood cell morphology examination of blood cells is very promising in clinical applications. Because it can significantly improve work efficiency, reduce the burden of manpower, avoid subjectivism, and facilitate standardization. The main difficulties lie in several key technical links, such as image acquisition, image segmentation, cell identification, and classification, etc. In recent years, both hardware devices and software algorithms have made rapid progress, which has led to the important development of artificial intelligence auxiliary systems from digital image acquisition, white blood cell segmentation, cell feature extraction, and classification. Compared with the traditional machine learning, the application of deep learning technology in the morphological identification of blood cells is particularly worthy of attention. In addition, the continuous emergence of microscopic blood cell image databases also provides important support for the further development and improvement of various algorithms. Understanding the key technical progress of artificial intelligence-assisted blood cell morphology examination will help to promote its continuous development and better clinical application. In recent years, artificial intelligence technology has changed from "traditional machine learning" to "deep learning", which no longer relies on manual extraction of features, but on its ability to automatically extract data to achieve. Compared with the blood cell image database from foreign countries, the construction of domestic databases should be strengthened to minimize the gap between foreign databases.
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Objective:To observe the pattern change of chemokine (C-X-C motif) ligand 12 (CXCL12)/C-X-C chemokine receptor type 4 (CXCR4) in systemic lupus erythematosus (SLE) by comparing the condition of SLE patients between pre-treatment and post-treatment.Methods:Peripheral blood mononuclear cells were obtained from 15 healthy controls and 8 first visit SLE patients before and after treatment. The expression of CXCR4, CD3, and CD19 on lymphocytes were detected by flow cytometry. CXCL12 concentration in serum was measured by enzyme-linked immunosorbent assay. For statistical analysis, the independent sample t test was used for independent samples of normally distributed data, the paired sample t test was used for paired samples of normally distributed data, the Mann-Whitney U test was used for independent samples of nonnormal distribution data, and the Wilcoxon test was used for comparison of related samples of non-normal distribution data. In the correlation analysis, Pearson correlation coefficient was used for data conforming to the normal distribution, and Spearman correlation coefficient was used for data not conforming to the normal distribution. Results:① The percentage of CXCR4 + lymphocyte subsets was significantly increased after treatment (90±7)% when compared with that before treatment (75±17)% ( t=-2.440, P<0.05). ② The mean fluorescence intensity of CXCR4 + lymphocyte subsets was significantly decreased after treatment [119.00(92.93, 142.50)] compared with that before treatment [177.00(130.75, 280.25)] ( Z=-2.100, P<0.05). ③ The plasma concentration of CXCL12 was significantly decreased after treatment (540±243) pg/ml when compared with that before the treatment (1 146±570) pg/ml ( t=3.219, P<0.05). ④ Moreover, there were significant correlations between the trends of the above three indicators after treatment. Conclusion:The proportion, the mean fluorescence intensity of CXCR4 + lymphocyte in peripheal blood subsets and the concentration of CXCL12 in plasma of first visit SLE patients are significantly changed after treatment. That suggests CXCL12/CXCR4 is closely associated with SLE.
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Objective To investigate the clinical features of rheumatoid arthritis combined with femur head necrosis. Methods The clinical data of 22 patients with rheumatoid arthritis combined with femur head necrosi were retrospectively analyzed. Results Among the 22 patients with rheumatoid arthritis combined with femur head necrosi, male was in 5 cases, female was in 17 cases, age was 28-69 (56.3 ± 1.9) years, and the disease duration of rheumatoid arthritis was 4-30 (14.1 ± 1.2) years. All patients had ≥ 2 grade femur head necrosis, with bilateral femur head necrosis in 9 cases, right femur head necrosis in 9 cases and left femur head necrosis in 4 cases. Only 2 cases did not take glucocorticosteroid, and the other 20 cases used glucocorticosteroid in the long-term. The treatment was no standardized. The multiple fracture was found in 4 cases by X-ray examination (the patients had no history of trauma); 4 cases examined bone density, and the results showed they all were osteoporosis. Conclusions Rheumatoid arthritis combined with femur head necrosis is not uncommon. The majority of patients receive long-term glucocorticosteroid treatment. Treatment is not standardized, and some patients are combined with osteoporosis.
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Objective To investigate the effects of Bucinnazine Hydrochloride on the pain behavior and the expression of caveolin-1 (Cav-1) in the anterior cingulate cortex of neuropathic pain mice.Methods 64 adult male Kunming mice (20-25g) were divided randomly into 4 groups with 16 in each group:Sham+BH(Bucinnazine Hydrochloride) group,Sham+NS (Normal Saline) group,CCI+ BH group and CCI+ NS group.The corresponding drugs were administered by intraperitoneal injectionfrom the forth day after CCI once a day for three days.Paw thermal withdrawal latency was measured by Hargreaves methods.Mechanicalwithdrawal threshold was assayed by electronic dolorimeter.c-Fos protein in anterior cingulate cortex was detected by immunohistochemistry staining and the expression of t-Cav-1,p-Cav-1was detected by Western blot.Results Bucinnazine Hydrochloride administered by intraperitoneal injection(0.1 mg/10 g,mice) alleviated thermal hyperalgesia and mechanical allodynia of CCI mice.Compared with the forth day (4.92±0.41) s of CCI+BH group,paw withdrawal latency on the fifth day(5.92±0.61) s was increased(P＜0.05),and on the sixth day(7.93±0.91) s and seventh day (9.12±0.69)s were increased more(P＜0.01,P＜0.01).The paw withdrawal mechanical threshold on the sixth and seventh day of CCI+BH group mice((2.54 ±0.41)g,(3.68±0.61)g) were increased significantly (P＜0.01,P＜0.01)compared with the forth day(1.55± 0.31)g.Immunohistochenistry results showed that the expression of c-Fos decreased after treated with Bucinnazine Hydrochloride in the anterior cingulate cortex of CCI mice(P＜0.001).Western Blotting showed that the expression of t-Cav-1 (1.97±0.31) and p-Cav-1 (0.11 ±0.09) in the anterior cingulate cortex of CCI +BH group mice decreased compared with that of in CCI+NS group mice(t-Cav-1:2.87±0.15,p-Cav-1:0.48± 0.09) (P＜0.01,P＜0.01).Conclusion Bucinnazine Hydrochloride can alleviate both thermal hyperalgesia and mechanical allodynia of neuropathic pain of mice,and reduce the expression of c-Fos,t-Cav-1,p-Cav-1 in the anterior cingulate cortex of neuropathic pain mice.
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Objective To identify the correlation between vitamin D prescription and recurrence in rheumatoid arthritis (RA).Methods A randomized controlled trial study was performed in 170 RA patients who were in remission during the past 2 months.According to the level of vitamin D,170 RA patients were divided into normal vitamin D group (84 patients) and deficiency vitamin D group (86 patients).Deficiency vitamin D group were randomly allocated to receive vitamin D treatment(vitamin D treatment group) or without vitamin D treatment (control group).In the 6-month follow-up period,the recurrence status was observed and compared.Results In the 6-month follow-up period,the recurrence rate of RA in normal vitamin D group was 16.7％(14/84),in vitamin D treatment group was 19.0％(8/42) and in control group was 29.5％(13/44),and there was no significant difference (P ＞ 0.05).In vitamin D treatment group,no hypercalcemia and hyperphosphatemia occured.The age,course of disease and remission time in recurrence patients of three groups were no significant difference (P ＞ 0.05).The level of vitamin D in recurrence patients of vitamin D treatment group was higher than that in recurrence patients of control group:(25.5 ± 8.9) ng/L vs.(20.9 ± 8.6)ng/L,and there was significant difference(P＜ 0.05).Conclusion Vitamin D deficiency is not identified to be a risk factor for RA recurrence.Vitamin D does not reduce the recurrence of RA.
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Objective To investigate the renal function damage in newberns ruth hyperbilirubinemia and the effects of early treatment. Methods 100 newberns with hyperbilirubinemia were taken as treatment group. Ser-um bilirubin, malondialdehyde (MDA), β_2-MG, and urine-minim protein (β_2-MG, α_1-MG, ALB) were measured within 24 hours after charge in. 50 healthy newborns had been chosen as controls. Anti-oxidate (vitamin E and Glu-tathion) as weel as regular method were given to the treatment group. All the above biochemical indexes were tested in the 3td and 6th day. Results When bilirubin level was in 205.0～256.5 μmol/L, β_2-MG、α_1 -MG in urine and MDA in blood were higher in treatment group than in control group(P < 0.05);when unconjugated bilirubin(UCB) was in in 256.6～342.0 μmol/L, β_2-MG was also raised (P < 0.01, P < 0.05);and when UCB greater than in 342.0 μmol/L, serum β_2-MG and urine ALB raised significantly (P <0.01,P <0.05). After early treatment, bil-irubin decreaed, and, β_2-MG, urine-minim protein declined to normal level, with a faster recovery in treatment group than control group (P < 0.05). Conclusions Hyperbilirubinemia may damage the glomerular filtration and renal tubule's re-absorption function in neonatals. Lipid peroxidation activated by UCB in Hyperbilirubinemia may cause thease damages. Antioxidant combined with regular treatment could lead better results.
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<p><b>OBJECTIVE</b>To investigate the anti-colon cancer effects of berberine and possible relationship with cyclooxygenase-2.</p><p><b>METHOD</b>Wistar rat colon cancer model was induced by 1-2 dimethylhydrazine (DMH) (40 mg x kg(-1), sc) + 1% dextran sodium sulfate solution (DSS) (freely drinking). All rats were randomly divided into 3 groups: Control (DMH + DSS + solvant), meloxicam (Mel) (DMH + DSS + Mel 1.35 mg x kg(-1)), berberine (Ber) (DMH + DSS + Ber 100 mg x kg(-1)). The drugs were given orally once a day for 5 day per week. The body weight, the number of colon ACFs, the incidence and number of colon cancer in rats, as well as the morphological changes of rat colon tissues were evaluated. Human colon cancer lovo cell line was treated by either Ber or Mel in various concentrations (1 10(-6) mol x L(-1), 1 x 10(-5) mol x L(-1), 1 x 10(-4) mol x L(-1), 1 x 10(-3) mol x L(-1)) for 6, 12 and 24 h, respectively, and the cell growth was assayed by MTT method. RT-PCR and western-blot were used to evaluate the mRNA and protein expressions of COX-2 from lovo cells treated with Ber and Mel.</p><p><b>RESULT</b>Ber significantly improved the dyscrasia induced by DMH + DSS, the both of body weight and general condition were better than control group. Ber also significantly inhibited ACF and colon cancer incidence in the rats treated by DMH + DSS for 10 weeks or 20 weeks, which was similar to that of Mel. Ber inhibited the proliferation of lovo cells in concentration- and time-dependent manners, and the IC50 values were significantly smaller than that of Mel at 6, 12 and 24 h after lovo cells were treated by either Ber or Mel. Ber also concentration-dependently decreased expressions of COX-2 mRNA and COX-2 protein from lovo cells.</p><p><b>CONCLUSION</b>Ber can inhibit ACF and tumor formation induced by DMH + DSS, and decrease the lovo cell proliferation index. The anti-tumor effects of Ber may involve in an unknown pathway through which the expressions of COX-2 mRNA and protein were inhibited.</p>
Subject(s)
Animals , Female , Male , Rats , 1,2-Dimethylhydrazine , Toxicity , Aberrant Crypt Foci , Berberine , Therapeutic Uses , Body Weight , Colonic Neoplasms , Cyclooxygenase 2 , Genetics , Dextran Sulfate , Toxicity , RNA, Messenger , Rats, WistarABSTRACT
Objective To investigate the value of deiminated recombinant rat filaggrin antibodies (ArFA)in predicting the activity of rheumatoid arthritis (RA).Methods ArFA was detected with enzyme linked immunosorbent assay(ELISA)in 61 patients with active RA and 48 patients with inactive RA.The ESR,CRP,RF were measured,while tender ioint count,swollen joint count and patient's general assessment were recorded.The level of ArFA in 20 RA patients was measured before and after treatment.Results The level of ArFA in patients with active RA was higher compared with patients with inactive RA [(130±35)U/L cs(66±25)U/L,P=0.004 ],the level of ArFA after treatment was reduced (79.8 U/ml vs 118.2 U/ml,P=0.000).Conclusion The level of ArFA can predict disease activity of RA.
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OBJECTIVE:To investigate the inhibition effect of berberine on colon carcinoma lovo cell. METHODS:Colon car-cinoma lovo cells were cultured and treated with berberine and meloxicam in various concentrations. At 6 h,12 h and 24 h after treatment,the proliferation of lovo cell was assayed by MTT method. RT-PCR method and Western blot testing were used to eval-uate the mRNA and protein expressions of COX-2 in lovo cell. RESULTS:Berberine with concentration larger than 10-5 mol?L-1 and meloxicam with concentration larger than 10-4 mol?L-1 could inhibit the growth and proliferation of lovo cell and showed concentration and time dependent manners(P
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Objctive To study the effects of traditional Chinese medicine S Kangdu mixture on functions of immune system and bone marrow in mice. Methods Animal model, which marrow and immune system were inhibited, was established by 60 Co or cyclophosphamide injection. The mixture was administrated orally. The white blood cell counts, spleen colony formed unites (CFU S), and the index of carbon clean up were measured. Results The mixture obviously improved leukopenia in peripheral induced by cyclophosphamide administration, and facilitated marrow karyocyte proliferation and formation of CFU S in 60 Co treated mice and reinforced phagocytosis of mononuclear phagocytic system in cyclophosphamide treated mice. Conclusion Traditional Chinese medicine S Kangdu mixture can significantly ameliorate immunological function and stimulate hemopoietic function in mice.
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Objective To establish a drug resistant human hepatoma cell line (QGY/DDP) induced by cisplatin (DDP) in vitro, and to investigate its biological features and mechanisms of resistance. Methods The drug resistant cell line (QGY/DDP) of hepatoma was established by intermittent administration of stepwise increas of the dosage of DDP. Drug sensitivity was evaluated by MTT assay. Cell counting was employed to graph the growth curve and to calculate the doubling time. Flow cytometry (FCM) was used to study the cell cycle distribution. In addition, the intracellular platinum accumulation was detected by atomic absorption spectrometry, and the expressions of P-glycoprotein (P-gp) and glutathione S-transferase-? (GST-?) were analyzed by FCM. Results QGY/DDP cell line was established successfully after 3 months with stable resistance to DDP, and the resistance index (RI) was 10.81. The established cell line exhibited obvious cross-resistance to 5-FU, VCR, MMC, EPI and HCPT. Compared with parental cell line, the QGY/DDP cell line grew slower and its doubling time became longer. The proportion of G0/G1 phase cells of the resistant cells was significantly higher than that of their parental cells, whereas the proportion of S and G2/M phase cells was decreased. The cellular platinum accumulation was obviously decreased in the QGY/DDP cell line, and the expression of GST-? in QGY/DDP cells was higher than that of their parental cells, but no over expression of P-gp was found in the resistant cell line. Conclusions QGY/DDP cell line shows the typical and stable resistant phenotype and possesses the basic biological features of resistant cells. The resistance of the cell line to DDP may be due to the reduction of intracellular platinum accumulation and the over expression of GST-?, but may not be related to the expression of P-gp.
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AIM: To observe the mRNA expressions of Insulin-induced gene 2(Insig-2) and Vitamin D receptor(VDR) in HepG2 cell line and investigate the possible mechanism of berberine on regulating blood lipid. METHODS: HepG2 cells were incubated with Bet at different concentrations(1,3,10,30,100 ?mol/L),and incubated with 3 ?mol/L berberine for various times(0,12,24,48,72 h).Total RNA was extracted and the mRNA expressions of Insig-2 and VDR from HepG2 cells treated by berberine were quantified by RT-PCR.(RESULTS): After exposure to gradient concentrations of berberine for 24 h,the mRNA expressions of Insig-2 and VDR in HepG2 cells increased obviously peaked at 3 ?mol/L concentration,then the up-regulating effect declined gradually.The relative coefficient between them was(0.753)(P
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Aim To construct a eukaryotic expression vector of human MCHR2 and to analyze the stable expression and signal transduction pathways of MCHR2 in CHO cells. Methods The full-length MCHR2 cDNA fragment was amplified by PCR from the human fetal brain cDNA library and inserted into eukaryotic expres-sion vector pcDNA3.1(+), resulted in the recombinant expression vector pcDNA3.1-MCHR2. The recombinant plasmid was confirmed by the restriction enzymatic digestion and DNA sequencing analysis, and then transfected into CHO cells by lipofectamine. A stably-transfected cell line was obtained by the dominant G418 selection, and the expression of the MCHR2 gene on transcription and translation levels were identified by RT-PCR and Western blot. Signal transduction pathways mediated by MCHR2 were analyzed by measurement of intracellular cAMP and calcium. Results The eukaryotic expression vector pcDNA3.1-MCHR2 was successfully constructed and the MCHR2 gene was stably transfected into CHO cells. A stably-transfected cell line was established and the MCHR2 gene was efficiently transcribed and translated. MCH stimulation had no effect on the production of cAMP, however, it could induce a clear and transient increase of intracellular calcium, suggesting that MCHR2 was only coupled to Gq protein. Conclusion The stable expression of MCHR2 and analysis of its signal transduction pathways in CHO cell line provided a solid experimental foundation for further studies on the function of the MCHR2 gene in vitro.
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Objective To construct eukaryotic expressing vector of human melanin-concentrating hormone receptor 1(MCHR1), then to transfect CHO cells with the vector for establishment of stable CHO cell line. Methods The full-length MCHR1 cDNA fragment was amplified by PCR from the human fetal brain cDNA library and then inserted into eukaryotic expression vector pcDNA3.1(+).The recombinant was transfected into CHO cells by lipofectamine TM 2000 after identification of digestion and sequencing on the recombinant eukaryotic expression vector pcDNA3.1(+)/MCHR1. The stable transfected CHO cell line was then established by screening cultures with G418, and the transcription and expression of MCHR1 were identified by RT-PCR, Western blot and immunofluorescence. Results The eukaryotic expression vector pcDNA3.1(+)/MCHR1 was constructed successfully, stable transfected CHO cell line was established, the MCHR1 protein was expressed successfully. Conclusion The construction of eukaryotic expression vector pcDNA3.1(+)/MCHR1 and the establishment of stable transfected CHO cell line provided a solid experimental foundation for further studies on the function of MCHR1.
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Objective To construct eukaryotic expression vectors expressing short hairpin RNA(shRNA)sections targeting human MCHR2 and to observe their effects on MCHR2 gene expression in HEK293 cell line.Methods According to the sequence of human MCHR2 gene,the oligonucleotides of shRNA were designed and synthesized and directionally cloned into plasmid pGenesil-1 with enhancing green fluorescence protein(EGFP)gene and Kan gene.The recombinant vectors were confirmed by enzyme digestion analysis and DNA sequencing.The recombinant vectors were transfected into HEK293 cell line by LipofectamineTM2000,the effects on MCHR2 at mRNA and protein levels were observed.Results Four shRNA expressing recombinants and the corresponding negative control vector were constructed and transfected into HEK293 cell successfully.MCHR2 transcript was reduced by about 45.8%-66.4%,the protein of MCHR2 was reduced by about 44.2%-81.0% in four transfectants respectively.Conclusion The construction of eukaryotic expression vectors expressing shRNA sections targeting human MCHR2 and identification successfully established a favourable foundation for further study on the function of MCHR2.