DNA barcoding method was conducted for the authentication of pollen materials due to difficulty of discriminating pollen materials bearing morphological similarity. In this study, a specific focus was to identify cattail pollen (Puhuang) and pine pollen (Songhuafen) samples from their adulterants which are frequently mixed-together. Regions of the internal transcribed spacer (ITS2) from 60 samples were sequenced, and new primers for cattail pollen were designed according to the sequence information. The results from the NJ trees showed that the species of pine pollen, Puhuang and their adulterants can be classified as obvious monophyly. Therefore, we propose to adapt DNA barcoding methodology to accurately distinguish cattail pollen, pine pollen and their adulterant materials. It is a great help for drug regulatory agency to supervise the quality of medicinal materials.
China ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Pinus ; classification ; genetics ; Pollen ; classification ; genetics ; Quality Control ; Typhaceae ; classification ; genetics

OBJECTIVETo investigate the possibility of transdifferentiation of adipose mesenchymal stem cells (AMSCs) into hepatocytes.

METHODSHuman omentum adipose tissue was dispersed with collagenase I. Cells collected were cultured in a DMEM-F12 medium containing 2% FBS supplemented with 20 ng/ml HGF, 10 ng/ml FGF4, 1xITS and 0.1 micromol/L dexasmison. The cells of the control group were also cultured in the same kind of medium but without any cytokines serving as a control. The expression of hepatic transcriptional factors such as GATA4 and HNF1 were checked by RT-PCR. At the end of the induction, hepatocyte markers were analysed by flow cytometry, and cytokeratin expressions were examined using cyto-immunofluorescence methods.

RESULTSAMSCs grew like fibroblasts and were passaged easily. Most of the third passaged AMSCs were positive against anti-CD29, anti-CD44 antibodies, but negative for the anti-CD34 and anti-CD45 ones. The hepatic transcriptional factor was expressed gradually to higher levels during the induction time. AFP and Alb positive cells were 30.0% and 17.8% of the total cultured cells, and the rate of cells positive to the two markers was 6.9%. The inducted cells were positive for CK18 and CK19 antibodies at the end of the induction. The cells in the control group were negative when checked by these methods.

CONCLUSIONSAMSCs could be directed to differentiate into hepatocytes in vitro by a cytokine cocktail with a low concentration FBS culture system.

Adipocytes ; cytology ; Cell Differentiation ; Cell Transdifferentiation ; Cells, Cultured ; Hepatocytes ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology

Objective To study the effect of different materials and porosities on bone formation in the scaffold after implantation of the degradable bone scaffold into human body. Methods According to natural reaction mechanism of fracture healing, the finite element method was used, combined with geometry of the scaffold, to establish a computationally coupled model based on material degradation curve and bone reconstruction control equation. Through this platform, representative volume elements of the scaffold with five kinds of materials and four types of porosities were selected for calculation and analysis, and dynamic process was reflected by bone mineral density (BMD) and maximum stress of the scaffold. Results The elastic modulus of the materials had a greater influence on growth of bone tissues in the scaffold. The smaller elastic modulus of the materials would lead to the greater amount of bone formation, but it would also have a greater impact on mechanical properties of the scaffold. The scaffold with higher porosity had lower rigidity, which could better promote formation of bone tissues, meanwhile it would also destroy mechanical stability of the scaffold. Conclusions According to performance requirements for different age, gender and location of bone tissues, personalized reference and calculation basis for selection of materials and porosity, structural design and clinical application of degradable porous bone scaffolds can be provided.

Objective: To evaluate clinical characteristics and prognosis of primary myelofibrosis (PMF) patients with thrombocytopenia in varied degrees. Methods: Clinical features and survival data of 1 305 Chinese patients with PMF were retrospectively analyzed. The prognostic value of thrombocytopenia in patients with PMF was evaluated. Results: 320 subjects (47%) presented severe thrombocytopenia (PLT<50×10(9)/L), 198 ones (15.2%) mild thrombocytopenia [PLT (50-99)×10(9)/L] and 787 ones (60.3%) without thrombocytopenia (PLT ≥ 100×10(9)/L). The more severe the thrombocytopenia, the higher the proportions of HGB<100 g/L, WBC<4×10(9)/L, circulating blasts ≥ 3%, abnormal karyotype and unfavourable cytogenetics (P<0.001, P<0.001, P=0.004, P<0.001 and P<0.001, respectively) were observed in this cohort of patients. The more severe the thrombocytopenia, the lower the proportion of JAK2V617F positive (P<0.001) was also noticed. Platelet count was positively correlated with splenomegaly, HGB and WBC (P<0.001, correlation coefficients were 0.131, 0.445 and 0.156, respectively). Platelet count was negative correlated with constitutional symptoms and circulating blasts (P=0.009, P=0.045, respectively; correlation coefficients were -0.096 and -0.056, respectively). The median survival of patients with severe thrombocytopenia, mild thrombocytopenia and without thrombocytopenia were 32, 67 and 89 months, respectively (P<0.001). Multivariate analysis identified thrombocytopenia in varied degrees (HR=1.693, 95%CI 1.320-2.173, P<0.001) and Dynamic Internation Prognostic Scoring System(DIPSS) prognostic model (HR=2.051, 95%CI 1.511-2.784, P<0.001) as independent risk factors for survival. Conclusion: PMF patients with severe thrombocytopenia frequently displayed anemia, leucopenia, circulating blasts and short survival, so active treatment measures should be taken especially in these patients.
Humans ; Primary Myelofibrosis ; Prognosis ; Retrospective Studies ; Thrombocytopenia

Objective: To explore the clinical implications and prognostic value of TP53 gene mutation and deletion in patients with myelodysplastic syndromes (MDS) . Methods: 112-gene targeted sequencing and interphase fluorescence in situ hybridization (FISH) were used to detect TP53 mutation and deletion in 584 patients with newly diagnosed primary MDS who were admitted from October 2009 to December 2017. The association of TP53 mutation and deletion with several clinical features and their prognostic significance were analyzed. Results: Alterations in TP53 were found in 42 (7.2%) cases. Of these, 31 (5.3%) cases showed TP53 mutation only, 8 (1.4%) cases in TP53 deletion only, 3 (0.5%) cases harboring both mutation and deletion. A total of 37 mutations were detected in 34 patients, most of them (94.6%) were located in the DNA binding domain (exon5-8) , the remaining 2 were located in exon 10 and splice site respectively. Patients with TP53 alterations harbored significantly more mutations than whom without alterations (z=-2.418, P=0.016) . The median age of patients with TP53 alterations was higher than their counterparts[60 (21-78) years old vs 52 (14-83) years old, z=-2.188, P=0.029]. TP53 alterations correlated with complex karyotype and International prognostic scoring system intermediate-2/high significantly (P<0.001) . Median overall survival of patients with TP53 alterations was shorter than the others[13 (95%CI 7.57-18.43) months vs not reached, χ(2)=12.342, P<0.001], while the significance was lost during complex karyotype adjusted analysis in multivariable model. Conclusion: TP53 mutation was more common than deletion in MDS patients. The majority of mutations were located in the DNA binding domain. TP53 alterations were strongly associated with complex karyotype and always coexisted with other gene mutations. TP53 alteration was no longer an independent prognostic factor when complex karyotype were occurred in MDS.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Genes, p53 ; Humans ; In Situ Hybridization, Fluorescence ; Middle Aged ; Mutation ; Myelodysplastic Syndromes/genetics* ; Prognosis ; Tumor Suppressor Protein p53 ; Young Adult

OBJECTIVETo study the effect of reserving articular capsule and traditional total hip replacement for old patients with unstable femoral neck fractures.

METHODSFrom January 2010 to January 2015, one hundred and twenty patients with femoral neck fracture were retrospective review. Among them, sixty patients were treated with total hip arthroplasty reserving articular capsule, the others were treated with traditional total hip arthroplasty without reserving articular capsule. The gender of reserving articular capsule group and traditional group were (male/female) 34/26, 31/29 respectively; the age were (73.4±4.4), (72.3±4.1) years old, respectively. All patients were followed up for six months, the operation time, blood loss, length of stay, incidence of postoperative complications, incidence of postoperative hip dislocation and hip Harris score of the two groups were observed.

RESULTSThe operative time of the reserving articular capsule group and traditional group were (95.68±6.90), (93.39±7.90) min (>0.05), and the blood loss were (998.78±15.20), (1 000.25±16.80) ml (>0.05). The time for hospital stay were (10.74±2.90), (13.25±2.20) days(<0.05). The Harris scores were 58.53±5.10, 49.38±4.90(<0.05) at 1 month after operation;91.08±7.50, 90.74±7.10(>0.05) at 6 months after operation.

CONCLUSIONSThe method of reserving auricular capsule can not increase the operative time and the blood loss. But it has a favorable effect on decreasing the hospital stay, complication, the rate of early dislocation of the hip joint, improving the function of hip joint.

BACKGROUNDExcessive iodine intake and viral infection are recognized as both critical factors associated with autoimmune thyroid diseases. Toll-like receptors (TLRs) have been reported to play an important role in autoimmune and inflammatory disorders. In this study, we aimed to clarify the possible mechanism of TLR3 involved in polyinosine-polycytidylic acid (poly(I:C)) promoting excessive iodine intake induced thyroiditis in non-obese diabetic (NOD) mice.

METHODSBoth NOD and BALB/c mice were randomly assigned to four groups: control group (n = 5), high iodine intake (HI) group (n = 7), poly(I:C) group (n = 7) and combination of excessive iodine and poly(I:C) injection (HIP) group (n = 7). After 8 weeks, mice were weighed and blood samples were collected. All the mice were sacrificed before dissection of spleen and thyroid gland. Then, thyroid histology, thyroid secreted hormone, expression of CD3(+) cells and TLR3 as well as inflammatory mRNA level were evaluated.

RESULTSBoth NOD and BALB/c mice from HI and HIP group represented goiter and increasing thyroid relative weight. Thyroid histology evidence indicated that only HIP group of NOD mice showed severe thyroiditis with lymphocytes infiltration in majority of thyroid tissue, severe damage of follicles and general fibrosis. Immunofluorescence staining results displayed a large number of CD3(+) cells in HIP NOD mice. Real-time polymerase chain reaction (PCR) results suggested interferon (IFN)-α increased over 30 folds and IFN-γ expression was doubled compared with control group, but interleukin (IL)-4 remained unchanged in HIP group of NOD mice thyroid. Meanwhile, over one third decrease of blood total thyroxine (TT4) and increased thyroid-stimulating hormone (TSH) was observed in HIP group of NOD mice. Only HIP group of NOD mice represented significantly elevation of TLR3 expression.

CONCLUSIONPoly(I:C) enhanced excessive dietary iodine induced thyroiditis in NOD mice through increasing TLR3 mediated inflammation.

Animals ; Female ; Inflammation ; chemically induced ; metabolism ; Iodine ; toxicity ; Mice ; Mice, Inbred NOD ; Poly I-C ; pharmacology ; Thyroiditis ; chemically induced ; immunology ; metabolism ; Toll-Like Receptor 3 ; metabolism

Halofuginone (HF) is a derivative of dichroine which is the extract of traditional Chinese medicine. It is widely used as an efficient anticoccidial drug. Recent studies have found that HF has unique biological activities, showing great potential capacities in the treatment of autoimmune diseases. In the article, we summarized the therapeutic effects of HF in a variety of autoimmune diseases and its mechanism, providing references for further clinical studies of HF.

OBJECTIVETo compare the clinical effects of isthmic spondylolisthesis by a single cage or double cages interbody fusion combined with pedicle screw fixation.

METHODSThe clinical data of 172 patients with isthmic spondylolisthesis underwent surgery from March 2000 to August 2008 were retrospectively analyzed. All cases underwent posterior pedicle screw fixation and interbody fusion, 89 cases with single cage fusion and 83 cases with double cages fusion. In single cage group, there were 56 males and 33 females, aged from 18 to 63 years old with an average of(41.60±8.20) years;25 cases were in L4 segment and 64 cases were in L5;according to the Meyerding standard, 32 cases were I degree, 46 cases were II degree and 11 cases were III degree. In double cage group, there were 49 males and 34 females, aged from 20 to 65 years old with an average of(43.30±6.39) years;21 cases were in L4 and 62 cases were in L5;according to the Meyerding standard, 25 cases were I degrees, 45 cases were II degree, and 13 cases were III degree. The operative time, intraoperative blood loss, postoperative drainage volum, bone fusion rate, intervertebral space height and the improvement of clinical symptoms were compared between two groups.

RESULTSAll the operations were successful and all patients were followed up for 18 to 83 months with an average of 4 years and 3 months. The operative time, intraoperative blood loss, postoperative drainage volume in single cage group were less than of double cage group(<0.05). Two weeks after operation, the intervertebral space height was significantly increased in two groups(<0.05), and there was no significant difference at the last follow-up between two groups. At 16 months after operation, all bone grafts of patients got bony fusion by X-rays. At the last follow-up, there were no statistically significant difference in JOA, ODI and VAS score between two groups, no pedicle screw loosening and breaking were found.

CONCLUSIONSSingle cage interbody fusion combined with pedicle screw fixation is as effective as with double cages interbody fusion in treatment of isthmic spondylolisthesis, it has the advantages of short operative time and less blood loss.

Objective:To observe the effect of the tactile vibration feedback system on the balance and walking function of the unilateral transitional amputees with prostheses. Methods:From October, 2019 to November 2020, eleven unilateral transtibial amputees with prostheses were selected from our hospital. They performed daily activities for three hours wearing tactile vibration feedback system designed by our department. The patients were evaluated by Performance Oriented Mobility Assessment (POMA), gait analysis and outdoor 1000 meters complex road walking test before and after treatment. Results:The balance score of the POMA improved (Z = -2.264, P < 0.05), the gait score and total score of the POMA increased significantly (|t| > 6.249, P < 0.001) after treatment. Gait analysis showed that the step length of the affected side improved, the bilateral step time and double support phase shortened, and the walking speed increased (|t| > 2.250, |Z| > 2.756, P < 0.05) after installation. The outdoor 1000 meters complex road walking test showed that the total steps, energy consumption and time decreased (|t| > 2.412, |Z| > 2.045, P < 0.05). All the patients finished the trail, and no discomfort appeared. Conclusion:The tactile vibration feedback system could safely and effectively improve the balance and walking function of the unilateral transtibial amputees with prostheses.