Background TLR-4 is a natural immunity receptors in immunity,and it plays an important role in the repair of central nervous system damage.But its effect in glaucoma optic nerve injury is unclear.Objective This study was to investigate the expression of TLR-4 in retina with high intraocular pressure(IOP)in genetic and Drotein level and therefore explore the mechanism of TLR-4 on retinal ganglion cells(RGCs)injury. Methods Chronic ocular hypertension models were established in the right eyes of 150 clean purebred Sprague-Dawley rats by cauterizing the 3 sallow sclera veins.IOP was measured before and after 2 h,1 day,3,7,14,28,56 days after operation by PEN Ⅱ TONO-type pen tonometer.The expression of TLR-4 protein in rat retina was detected by immunohistochemistry and Western blot,and expression of TLR-4 mRNA was assayed by real time-PCR.This experimental procedure foliowed the Statement of Association for Research in Vision and Ophthalmology. Results The IOP was elevated in various time points after operation in experimental group,showing significant differences in comparison with control group(P<0.01).The immunohistochemistry revealed that the expression of TLR-4 protein in rat retina with chronic hypertension in 2 h,1 day,3,7,14,28,56 days after operation with the high A298 values in comparison with control eyes(P<0.05-0.01).Increased levels of TLR-4 mRNA in rat retinas were detected by RTPCR in high IOP eyes compared with control eyes in all time points after operation,presenting statistically significant differences between two groups(P<0.05-0.01).Western blot detection displayed the high expression of TLR-4 in retina in high IOP eyes early after operation with statistically significant results between model group and control group (P<0.05-0. 01). Conclusion TLR-4 is up-regulated in rat retina with chronic high IOP,suggesting that TLR-4 plays an immunoregulatory effect in glaucomatous eye.

AIMTo investigate the tissue distribution and pharmacokinetics of oridonin-solid lipid nanoparticles in animals.

METHODSHPLC method was established to determine the concentration of oridonin in serum of rabbits and in different tissues of mice. The results after tail iv administration of oridonin and oridonin solid lipid nanoparticles were compared.

RESULTSThe relative tissue content of oridonin of solid lipid nanoparticles in the liver, spleen, lung, heart and kidney were 4.25%, 3.44%, 1.19%, 0.52% and 0.60%, respectively. The concentration-time curves of oridonin and oridonin solid lipid nanoparticles were both fitted to the three-compartment model. T(1/2)pi = 0.087 h, T(1/2)alpha = 1.65 h, T(1/2)beta = 32.36 h, V(C) = 0.66 mL.kg(-1).

CONCLUSIONSolid lipid nanoparticles could increase the hepatic and lienic targeting efficiency of oridonin in mice and improve its bioavailability. Solid lipid nanoparticles were helpful for oridonin to reach a long circulation time and were hopeful to be its novel drug carrier.

Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; isolation & purification ; pharmacokinetics ; Area Under Curve ; Diterpenes ; administration & dosage ; isolation & purification ; pharmacokinetics ; Diterpenes, Kaurane ; administration & dosage ; isolation & purification ; pharmacokinetics ; Drug Carriers ; Drug Delivery Systems ; Female ; Injections, Intravenous ; Isodon ; chemistry ; Lipids ; Liver ; metabolism ; Male ; Mice ; Nanoparticles ; Plants, Medicinal ; chemistry ; Rabbits ; Spleen ; metabolism ; Tissue Distribution

OBJECTIVES: This study aims to evaluate the effectiveness of the Lean Six Sigma approach in improving procedure for (TAT) of reverse transcriptase polymerase chain reaction (RT-PCR) for SARS-CoV-2 testing at The Medical City. Specific objectives of the study are to determine the following: 1) baseline sigma and average TAT (in hours); 2) post-implementation sigma and average TAT (in hours) 3) compare if there is a significant improvement between baseline and post-implementation sigma and average TAT (in hours) 4) effect on workflow efficiency. METHODOLOGY: Lean Six Sigma method for quality improvement was applied using DMAIC: Define, Measure, Improve, and Control. The root causes identified were lack of manpower, equipment, space, and manual and complex processes. Then, process wastes were identified, and corresponding proposed solutions were sustained in the control phase, such as standardization and the use of automation. Measurement of turn-around time and six sigma of the process were performed for evaluation. RESULTS: Results showed a significant improvement in the TAT in RT-PCR results, with most results released within 24 hours. The pre-Lean Six Sigma data on TAT were as ollows: 24.88% released within 24 hours; 65.14% released within 24-48 hours; 3.56% released within 48-72 hours, and 6.42% released in more than 72 hours. The post Lean Six Sigma TAT were as ollows: 95.32% released within 24 hours; 4.29% released within 24 to 48 hours; 0.13% released within 48-72 hours, and 0.12% released more than 72 hours. The computed sigma post-implementation was increased from 3.56 to 4.82. The p-value was calculated using the chi-square test, and the computed chi-square statistic is 1894.1021. The p-value is <0.00001 and the result is significant at p<.05. Although there is a significant decrease in the volume of samples post implementation due to the changing COVID-19 situation, real time TAT was improved. It also resulted to increased workflow efficiency with the use of lesser manpower with more appropriate utilization. CONCLUSION Applying the Lean Six Sigma method to improve quality processes in the laboratory is shown to be practical, cost-effective, and straightforward.
Lean Six Sigma ; SARS-CoV-2

Objective To observe the influences of Ganfujian Formula on the expressions of transforming growth factor-β1 (TGF-β1), TβRII-mRNA, Smad2/3, Smad4 and Smad7 in rats with hepatic fibrosis induced by porcine serum.Methods The hepatic fibrosis model was established in SD rats by the intraperitoneal injection of porcine serum (0.5 mL, twice a week for 8 weeks). The rats then were given Ganfujian Formula orally in different doses (1.50 g/kg, 2.25 g/kg and 3.0 g/kg) and colchicines (0.154 g/kg) respectively until the end of 12th week. The expressions of TGF-β1 and TβRII-mRNA were detected by using RT-PCR and the expressions of Smad2/3, Smad4 and Smad7 were detected by applying Western blotting method.Results In the model group, compared with the normal control group, the expressions of TGF-β1 and TβRII-mRNA, Smad2/3 and Smad4 increased significantly and Smad7 expression decreased significantly. After the treatment with Ganfujian Formula the expressions of TGF-β1 and TβRII-mRNA were reduced in the treatment group compared with the model group. Ganfujian Formula regulated downward the expressions of Smad2/3 and Smad4, and regulated upward Smad7 expression at the same time.Conclusion Ganfujian Formula can inhibit the expressions of TGF-β1 and TβRII-mRNA, regulates downward the expressions of Smad2/3 and Smad4, and regulates upward Smad7 expression. It may be its one of the mechanisms for curing hepatic fibrosis that it has an obvious influence on hepatic TGF-β and Smad signal pathway.

Objective: To investigate the value of next-generation sequencing (NGS) technology in the prognosis monitoring and treatment guidance for molecular minimal residual disease (MRD) in acute myeloid leukemia (AML) patients with complete remission (CR). Methods: The clinical data of 68 AML (non-acute promyelocytic leukemia) patients who received gene mutation spectrum by using NGS technology at initial diagnosis and in CR phase in Tangdu Hospital of Air Force Military Medical University from January 2016 to July 2018 were retrospectively analyzed. The recurrence and survival of both molecular MRD positive group and negative group were analyzed and compared, and the value of NGS technology and multiparameter flow cytometry (MFC) were also analyzed in MRD monitoring. Results: There were 39 males (57.4%) and 29 females (42.6%) in 68 patients, and the median age was 52 years old (8-82 years old). Molecular MRD positive group included 38 patients, while negative group included 30 patients. Residual mutation gene type in CR phase was most frequently detected in epigenetic regulator gene mutations, such as ASXL1, TET2, DNMT3A and IDH1/IDH2. Statistical analysis showed that the 2-year cumulative recurrence rate (CIR) in the molecular MRD positive group was higher than that in the molecular MRD negative group (86.8% vs. 51.3%; χ² = 9.249, P = 0.002); the 2-year relapse-free survival (RFS) rate in the molecular MRD positive group was lower than that in the molecular MRD negative group (13.2% vs. 48.7%; χ² = 9.249, P = 0.002); the 2-year overall survival (OS) rate in the molecular MRD positive group was lower than that in the molecular MRD negative group (58.0% vs. 100%; χ² = 4.122, P = 0.042). Up to follow-up date, 3 patients with molecular MRD positive and 1 patient with molecular MRD negative who received allogeneic hematopoietic stem cell transplantation (allo-HSCT) were still in disease-free survival. The results of monitoring MRD showed high consistency (76.7%, 33/43) in NGS and MFC. Compared with the other groups, the patients with both positive NGS and MFC had a higher relapse rate, and the difference was statistically significantly (P < 0.05). Conclusions Molecular MRD of AML patients is detected by using NGS technology, which could be used to predict the relapse and survival, suggesting that molecular MRD may guide post-remission treatment regimens and the determination of allo-HSCT indications.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; DNA (Cytosine-5-)-Methyltransferases ; genetics ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Mutation ; Sequence Analysis, DNA ; Young Adult

OBJECTIVE: To investigate the clinical outcomes and prognostic factors of refractory/relapsed acute myeloid leukemia (AML) patients who received allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: The clinical data of 80 refractory/relapsed AML patients who received allo-HSCT from December 2013 to June 2020 were retrospectively analyzed, including the overall survival (OS) rate, disease-free survival (DFS) rate, relapse rate, incidence of transplant-related mortality (TRM), and the related risk factors were explored. RESULTS: Hematopoietic reconstitution was obtained in all 80 patients after transplantation, the 3-year OS and DFS rates were (48.8±6.3)% and (40.8±6.7)%, respectively. The 3-year cumulative incidence of relapse and TRM were 33.8% (95%CI: 0.254-0.449) and 15.0%(95%CI: 0.114-0.198), respectively. Univariate analysis showed that non-remission (NR) status before transplantation, DNMT3A R882 mutations and grade II-IV acute graft-versus-host disease (aGVHD) had negative effects on OS and DFS. Multivariate analysis indicated that the DNMT3A R882 mutations and grade II-IV aGVHD were independent risk factors for OS (HR=0.253, 95%CI: 0.092-0.695, P=0.008; HR=5.681, 95%CI: 2.101-15.361, P=0.001) and DFS (HR=0.200, 95%CI: 0.071-0.569, P=0.003; HR=7.117, 95%CI: 2.556-19.818, P<0.001). The 3-year cumulative incidence of relapse was 71.4%(95%CI: 0.610-0.836) in genetic high-risk group, which was higher than 23.3%(95%CI: 0.147-0.370) in intermediate-risk group and 23.5%(95%CI: 0.127-0.437) in favorable-risk group (P=0.006). CONCLUSION Allo-HSCT is an effective and safe choice for refractory/relapsed AML patients. DNMT3A R882 mutations and grade II-IV aGVHD are negative prognostic factors of allo-HSCT for refractory/relapsed AML patients.
Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation/adverse effects* ; Humans ; Leukemia, Myeloid, Acute/therapy* ; Prognosis ; Recurrence ; Retrospective Studies ; Transplantation, Homologous/adverse effects*

Background:The increasing prevalence of colorectal cancer (CRC) in China and the paucity of information about relevant expenditure highlight the necessity of better understanding the financial burden and effect of CRC diagnosis and treatment.We performed a survey to quantify the direct medical and non-medical expenditure as well as the resulting financial burden of CRC patients in China.Methods:We conducted a multicenter,cross-sectional survey in 37 tertiary hospitals in 13 provinces across China between 2012 and 2014.Each enrolled patient was interviewed using a structured questionnaire.All expenditure data were inflated to the 2014 Chinese Yuan (CNY;1 CNY =0.163 USD).We quantified the overall expenditure and financial burden and by subgroup (hospital type,age at diagnosis,sex,education,occupation,insurance type,household income,clinical stage,pathologic type,and therapeutic regimen).We then performed generalized linear modeling to determine the factors associated with overall expenditure.Results:A total of 2356 patients with a mean age of 57.4 years were included,57.1％ of whom were men;13.9％ of patients had stage Ⅰ cancer;and the average previous-year household income was 54,525 CNY.The overall average direct expenditure per patient was estimated to be 67,408 CNY,and the expenditures for stage Ⅰ,Ⅱ,Ⅲll,and Ⅳ disease were 56,099 CNY,59,952 CNY,67,292 CNY,and 82,729 CNY,respectively.Non-medical expenditure accounted for 8.3％ of the overall expenditure.The 1-year out-of-pocket expenditure of a newly diagnosed patient was 32,649 CNY,which accounted for 59.9％ of their previous-year household income and caused 75.0％ of families to suffer an unmanageable financial burden.Univariate analysis showed that financial burden and overall expenditure differed in almost all subgroups (P ＜ 0.05),except for sex.Multivariate analysis showed that patients who were treated in specialized hospitals and those who were diagnosed with adenocarcinoma or diagnosed at a later stage were likely to spend more,whereas those with a lower household income and those who underwent surgery spent less (all P ＜ 0.05).Conclusions:For patients in China,direct expenditure for the diagnosis and treatment of CRC seemed catastrophic,and non-medical expenditure was non-ignorable.The financial burden varied among subgroups,especially among patients with different clinical stages of disease,which suggests that,in China,CRC screening might be cost-effective.