OBJECTIVE:To compare the contents of 2 index components in salt frying and salt steaming samples of Psoralea corylifolia,and select better processing technology. METHODS:Under the conditions that ratio of salt and medicinal materials was 1:50,4-5 times salt of adding water,brine run time was 2 h,the amounts of psoralen and isopsoralen in salt frying samples after 10,12,14 min of frying under 150 ℃,170 ℃,190 ℃ and in salt steaming samples after 1,1.5,2,2.5 h of steaming were re-spectively investigated,and the effects of 2 processing technology were compared. RESULTS:Under fixed condition,the contents of index components was the highest in salt frying samples by fried for 12 min under 150 ℃ and in salt steaming samples by steamed for 1 h;the content in salt steaming samples (1.49%) was higher than that in salt frying samples (1.29%)(P=0.011). CONCLUSIONS:According to the comparison of index components'contents in processing samples,salt steaming is superior to salt frying of P. corylifolia.

Objective To discuss the value of CR and CT to diagnose early ankylosing spondylitis. Methods Clinical data and the signs of CR and CT of eighteen cases which were diagnosed as early ankylosing spondylitis were analyzed. Results Both CR and CT imaging could show the diseased region, morphous change, and the extent of diseased joint. CT imaging could show the tiny change of diseased articular facet. Conclusion CR plain is the first choice to diagnose early ankylosing spondylitis. As doubtful case, CR combination with CT scan can raise the accuracy rate of diagnosis to early ankylosing spondylitis.

Objective To construct bone marrow stem cell sheets and to investigate its effects in the process of osteogenesis.Methods BMSCs were differentiated into osteoblasts and then seeded into a temperature responsive culture dish to construct BMSC sheets.PLGA scaffolds in which both BMSC suspension and BMSC sheets were added,were implanted into the left side of the dogs' mandible.In the other side,PLGA scaffolds that were not wapped with BMSC sheets were implanted as control.At 16 weeks,the samples were processed for radiological analysis and histological examination.Results Cells in the BMSC sheets grew well.In the experimental side,the optical density of the samples was higher than that of the control side (P＜0.05) and plenty of lamellar bones and Haversian system were observed.Conclusions The formation of lamellar bones can be promoted by PLGA scaffolds and BMSC sheets in the process of tissue engineering bone reconstrution.

Objective To evaluate the effect of oxycodone on migration of human colon cancer cells and the role of μ and κ receptors.Methods The human colon cancer HCT116 cells at the logarithmic growth phase were seeded in 24-well or in 6-well plates at a density of 1 × 106 cells/mnl (0.5 ml/well or 2 ml/well,144 wells in total).The cells were divided into 6 groups (n=24 each) using a random number table:control group (group C),1,5 and 10 μmol/L oxycodone groups (group O1,group O2 and group O3),oxycodone plus μ receptor antagonist CTOP group (group O2+CTOP) and oxycodone plus κ receptor antagonist nor-binaltorphimine group (group O2+BNI).The cells were incubated for 24 h with oxycodone 1,5 and 10 μmol/L in O1,O2 and O3 groups,respectively.The cells were incubated for 24 h with 5 μmol/L oxycodone plus 20 μmol/L CTOP and 5 μmol/L oxycodone plus nor-binahorphimin 20 μmol/L in O2+CTOP and O2+BNI groups,respectively.The invaded and migrated cells were counted,and the levels of Ras homolog gene family member A (RhoA),Rho-associated protein kinase 1 (ROCK1),matrix metalloproteinase-2 (MMP-2) and MMP-9 were detected.Results Compared with group C,the number of invaded and migrated cells was gradually decreased,and the levels of RhoA,ROCK1,MMP-2 and MMP9 were gradually decreased in O1,O2 and O3 groups (P＜0.05),and no significant change was found in the parameters mentioned above in group O2+BNI (P＞0.05).Compared with group O2,the number of invaded and migrated cells was significantly increased,and the levels of RhoA,ROCK1,MMP-2 and MMP9 were increased in group O2 + BNI (P＜0.05),and no significant change was found in the parameters mentioned above in group O2+CTOP (P＞0.05).Conclusion Oxyc odone can inhibit the migration of human colon cancer cells,and the mechanism is totally related to inhibition of RhoA/ROCKl signaling pathway activation after activating κ receptors,but not related to μ receptors.

Objective To investigate the expression of macrophage migration inhibition factor (MIF) and cell cycle regulating factor Cyclin D1 in hepatocellular carcinoma tissue and the interaction between MIF and Cyclin D1 in hepatocellular carcinoma cell cycle controlling. Methods Using quantitative real-time PCR and Western blotting to detect mRNA and protein expression of MIF and Cyelin DI in HCC tissues and tumor adjacent tissues. Specific small interfering RNA(siRNA) targeting MIF gene was transfccted at doses of 50 nmol/L and 100 nmoL/L into HCC cell lines of PLC and HepG2 with lipofeetamine 2000 methods to knockdown the expression of M1F gene and to investigare the the interaction between M1F and Cyclin D1. Results MIF and Cyclin D1 protein and mRNA were overexpressed in HCC tumor tissues. The relative expression of MIF,Cyclin D1 protein and mRNA were 0.825±0.13,0.843± 0.104 and 7.31±1.85 folds、4.27±1.05 folds, compared with the tumor adjacent tissues (FMIF= 15.5, P<0.01;FCyclin D1=87.5,P <0.01). In MIF siRNA treated PLC and HepG2 cells, MIF mRNA down regulation 71.2%±7.2%, 87.4%±2.9% ,74.3%±8.9% and 88.4%±4.6% respectively (FPLC = 315.5 ,P < 0.01 ; FHepG2= 201.2 P < 0.01). While MIF protein expression were significandy reduced to 0.33±0.03,0.11±0.02, 0.81±0.08 and 0.36±0.02 in a dose-dependent manner (FPLC= 43.9, P <0.01 ;FHepG2 = 133.4 P <0.01). Cyclin D1 mRNA was significantly down-regnlated in MIF siRNA treated PLC and HepG2 cell lines when compared with control group(P <0.01). In 50 nmol/L and 100 nmol/L groups, Cyclin DI mRNA levels were respectively decreased by 68.2%±3% and 78.1%±1.4% in PLC cell, 65.8%±4.7% and 77.3%±2.6% in HepG2 cell (FPLC= 1569, P < 0.01 ; FHepG2= 480.4, P <0.01). Compared with control groups, Cyclin D1 protein levels significantly reduced to 0.28±0.06、0.15±0.03 and 0.44 ±0.04、0.13±0.02 in the PLC and HepG2 after M IF siRNA treatment(FPLC= 35.5, P < 0.01 ; FHepG2 = 114.7, P < 0.01). Conclusions MIF and Cyclin D1 mRNA and protein were overexpressed in HCC tumor tissues and participated in tumor cell cycle regulation. MIF may up-regnlate the expression of Cyclin DI via ERK signalling and precipitate in carcinogenesis of hepatocellular carcinoma.

OBJECTIVETo detect potential mutation of the UGT1A1 gene in a child affected with Crigler-Najjar syndrome type II.

METHODSBlood samples were collected from the patient and his parents for the extraction of genomic DNA. Potential mutation of the UGT1A1 gene was detected with polymerase chain reaction (PCR) and direct sequencing. The child was followed up until the age of 3 years and 6 months.

RESULTSThe patient showed persistent unconjugated hyperbilirubinemia. Sequencing of the UGT1A1 gene has detected a rare heterozygous c.610 A>G (p.Met204Val) mutation in the exon 1, in addition with a heterozygous c.1091 C>T (p.Pro364Leu) mutation in exon 4. The two mutations were inherited from his father and mother, respectively. The patient was diagnosed with Crigler-Najjar syndrome type II and received oral phenobarbital treatment.

CONCLUSIONThe compound UGT1A1 gene mutation probably accounts for the disease in the patient manifesting persistent mild unconjugated hyperbilirubinemia. Genetic counseling and prenatal diagnosis should be provided for his family.

Crigler-Najjar Syndrome ; genetics ; Glucuronosyltransferase ; genetics ; Humans ; Infant ; Male ; Mutation ; Sequence Analysis, DNA

OBJECTIVETo reconstruct mandibular defect using tissue engineering bone with bone marrow stem cells (BMSCs) cell sheets and investigate the effect of cell sheets on osteogenesis.

METHODSBMSCs were isolated with the method of density gradient centrifugation from canine and cultured. BMSCs were induced to differentiate to osteoblasts. BMSCs induced were fabricated to BMSCs cell sheets. The poly (lactic-co-glycolic acid) (PLGA) wrapped with cell sheets were implanted into the mandibular defect in the left side (experimental side). PLGA wrapped without cell sheets were implanted into the right side (control side) of mandibles. 16 dogs were evenly divided into 4 groups, and one group of them was executed in 4, 8, 12, 16 weeks for gross investigation and histological observation.

RESULTSThe osteogenesis of experimental side was better than that of control side. 16 weeks after implantation, most areas of the mandibular defect were replaced by fresh bone tissue. Compact bone similar to normal bone tissue formed in the lingual defect of mandible and had bony union with the bone stump. The optical density of the fresh bone in the experimental side was higher than that of the control side, there was a significant difference between the two methods (P<0.05). Plenty of lamellar bones formed in experimental side and Haversian system, as well as red marrow, were observed.

CONCLUSIONTissue engineering bone with the structure of lamellar bones can be formed by the technology of BMSCs cell sheets.

Animals ; Bone Marrow Cells ; Bone and Bones ; Dogs ; Lactic Acid ; Mandible ; Osteoblasts ; Osteogenesis ; Polyesters ; Polyglycolic Acid ; Polymers ; Tissue Engineering

Objective:To study the clinical characteristics and differences of severe hyperbilirubinemia caused by hemolytic disease of the newborn (HDN) and glucose-6-phosphate dehydrogenase (G6PD) deficiency.Methods:From January 2014 to December 2021, newborns (gestational age ≥ 35 weeks and postnatal age ≤ 28 d) admitted to the Department of Neonatology of Hunan Children's Hospital with severe hyperbilirubinemia caused by HDN or G6PD deficiency were retrospectively analyzed. According to the etiology of hyperbilirubinemia, they were assigned into HDN group and G6PD deficiency group. The general conditions, clinical manifestations, laboratory results, treatment and prognosis of the two groups were compared using SPSS 26.0 software.Results:A total of 532 cases were in the HDN group and 413 cases in the G6PD deficiency group. The HDN group reached peak hyperbilirubinemia earlier than the G6PD deficiency group [3(2,5) d vs. 6(4,8)d, P<0.05]. The HDN group had lower peak value of total serum bilirubin [379.5(345.6,426.7) μmol/L vs. 486.4 (413.5,577.4) μmol/L] and lower incidence of anemia [37.4% (199/532) vs. 55.0% (227/413)]than the G6PD deficiency group.The incidence of anemia with elevated reticulocyte percent(Ret%) in the HDN group was higher than the G6PD deficiency group[66.3%(132/199) vs. 5.7%(13/227), P<0.05]. Compared with the G6PD deficiency group, the incidences of exchange transfusion and repeated (≥2 times) exchange transfusion, acute bilirubin encephalopathy(ABE) and the mortality rate after withdrawal of treatment in the HDN group were significantly lower ( P<0.05). Conclusions:Neonatal severe hyperbilirubinemia caused by HDN has early onset. G6PD deficiency caused hyperbilirubinemia has higher incidences of anemia, more severe jaundice and ABE, without increased Ret%.

Objective To investigate the correlation between neutrophil-lymphocyte ratio (NLR) and disease activity of systemic lupus erythematosus (SLE),and the changes of NLR in different organ involvement of SLE patients.Methods A total of 155 SLE patients and 135 healthy controls from the Rheumatology Department of Xiangya Hospital were enrolled in this study from 2010 to 2018.Patients with SLE were divided into lupus nephritis group (LN group) and non-lupus nephritis group (non-LN group),serositis group and non-serositis group,according to whether they had kidney involvement or serositis.According to the SLE disease activity index 2000(SLEDAI-2000),the patients were divided into mild to moderate disease activity group (SLEDAI score ＜ 15) and severe disease activity group (SLEDAI score≥ 15).The NLR values of the above groups were compared.Spearman's correlation analysis was used to analyze the correlation between NLR and SLE patients' laboratory indexes.Multiple linear regression model was used to analyze the relationship between NLR and SLE disease activity.Receiver operating characteristic curve (ROC) was used to evaluate the value of NLR in SLE diagnosis and activity assessment.Results (1)The NLR value of SLE patients was significantly higher than that of healthy control group,and the difference was statistically significant (P ＜ 0.01).(2)The NLR value of SLE patients in the LN group was higher than that in the non-LN group,and the NLR value of SLE patients with serositis was higher than that in the group without serositis,with statistically significant differences (both P ＜ 0.05).(3)The NLR value of SLE patients in the severe disease activity group was higher than that in the mild and moderate disease activity group,and the difference was statistically significant (P ＜ 0.01).(4)NLR of SLE patients was positively correlated with CRP (rs=0.188,P=0.019),SLEDAI score (rs=0.264,P=0.001),and negatively correlated with total serum protein (rs=-0.250,P=0.002) and serum albumin (rs=-0.329,P ＜ 0.001),respectively.(5) Multiple linear regression showed that NLR was independently associated with SLE disease activity (B=0.351,95％CI 0.012-0.690,t=2.047,P=0.042).(6) According to ROC curve,the optimal cut-off value of NLR for SLE diagnosis was 2.17 (sensitivity 60.0％,specificity 83.1％,AUC=0.744),and the best cut-off value for predicting the activity of severe disease activity in SLE patients was 3.28 (sensitivity 58.5％,specificity 78.1％,AUC=0.700).Conclusion NLR is closely related to renal involvement,serositis and disease activity in SLE patients,which indicates that NLR,as a new inflammatory indicator,is of great significance for the assessment of SLE disease activity and organ involvement.

BACKGROUND:Previous research by the research team found that domestically produced porous tantalum is beneficial for early adhesion and proliferation of MG63 cells,and can be used as a scaffold material for bone tissue engineering. OBJECTIVE:To investigate the effect of domestic porous tantalum modified by osteogenic induction factor slow-release system on the adhesion,proliferation,and differentiation of MG63 cells. METHODS:Osteogenic induction factor slow-release system was constructed by adding 15%volume fraction of osteogenic factor solution to poly(lactic-co-glycolic-acid)gel.The passage 3 MG63 cells were inoculated on a porous tantalum surface(control group),porous tantalum surface coated with poly(lactic-co-glycolic-acid)copolymer gel(gel group),and porous tantalum surface coated with osteoblastic induction factor slow-release system(slow-release system group),and co-cultured for 5 days.The surface cytoskeleton of the material was observed by phalloidine staining.Cell proliferation was detected by flow cytometry.Western blot assay and RT-qPCR were used to detect the protein and mRNA expressions of type Ⅰ collagen,osteopontin,and RUNX-2 on the surface cells of the material. RESULTS AND CONCLUSION:(1)Phalloidine staining showed that MG63 cells adhered to and grew on the surface and inside of the three groups of porous tantalum,and the matrix secreted by the cells covered the surface of the material.(2)Flow cytometry showed that the cell proliferation in the slow-release system group was faster than that in the control group and the gel group(P<0.05).(3)Western blot assay and RT-qPCR showed that the protein and mRNA expressions of type Ⅰ collagen,osteopontin,and RUNX-2 in the slow-release system group were higher than those in the control group and gel group(P<0.05).(4)The results showed that the domestic porous tantalum modified by the osteogenic induction factor slow-release system was beneficial to the adhesion,proliferation,and differentiation of MG63 osteoblasts.