51 cases of osteoporosis were put on clinical trial in our OPD and randomly assigned to TCM treatment group (N = 17), TCM control group and blank group. The total effective rate in the TCM treatment group obtained was 94. 1% and the significantly effective rate achieved 88. 2%. After the treatment, the density of the osseous tissue increased by 3-4%, main symptoms improved markedly, the serum calcium increased and the urine pyridine phenol decreased. All the findings were superior than that in the rest two groups. This composite is effective and safe (no side reactions) and worth put into large sale production.

Objective: To explore the effect of atorvastatin (Atv) on high glucose-induced oxidative stress injury in human umbilical vein endothelial cells (HUVECs) by SIRT1/NADPH oxidase pathway with the possible mechanisms.
Methods: HUVECs were cultured in low glucose medium and then divided into 6 experimental groups:①Normal group,②Osmotic pressure control group,③High glucose (HG) group,④HG+Atv (0.1, 1.0, 10.0)μmol/L group,⑤HG+sirtinol (SIRT1 inhibitor) group,⑥HG+apocynin (NOX4 inhibitor) group, and HUVECs were further cultured for 24 hours. The cell proliferation was examined by CCK-8 kit, ROS level was detected by lfow cytometry method, protein expressions of SIRT1 and NOX4 were measured by Western blot analysis.
Results: ① Compared with Normal group, HG group had decreased HUVECs proliferation, Atv improved the HG inhibited proliferation in a does dependent manner. ② HG group had the higher level of ROS, increased NOX4 protein expression and decreased SIRT1 protein expression. ③ In HG condition, Atv up-regulated SIRT1 expression and down-regulated ROS and NOX4 expressions in a does dependent manner.④In HG condition, sirtinol decreased SIRT1 expression, increased NOX4 expression, and apocynin decreased NOX4 expression, while it had no inlfuence on SIRT1 expression.
Conclusion: Atorvastatin could resist HG-induced oxidative stress injury in HUVECs, which might be related to up-regulated SIRT1 expression, and SIRTI plays the role in NADPH oxidase at upstream.

BACKGROUND:The induced concentration for osteoblasts is often introduced as reference to induce odontoblast differentiation. However, there are no reports on other concentrations. OBJECTIVE: To observe the expression of dentin matrix protein-1, dentin sialoprotein and matrix extracelular phosphoglycoprotein during low-dose β-glycerophosphate-induced differentiation of dental pulp stem cels into odontoblasts. METHODS:Human dental pulp stem cels were isolated and cultured, and then induced by different concentrations of inducing solution to differentiate into adipocytes and osteoblasts, which could verify the multi-directional differentiation ability of human dental pulp stem cels. Under 5 mmol/L β-glycerophosphate, dental pulp stem cels differentiated into odontoblasts. At 7, 14, 21, 28 days of culture, RNA samples were extracted from dental pulp stem cels in each group, and reverse-transcription PCR was used to detect the expression of dentin matrix protein-1, dentin sialoprotein and matrix extracelular phosphoglycoprotein. Mineralized nodules were detected by alizarin red S staining to show the successfuly osteogenesis induction. RESULTS AND CONCLUSION: Human dental pulp stem cels could be induced to adipocytes and osteoblasts. The results of reverse-transcription PCR showed that the dental pulp stem cels under 5 mmol/L β-glycerophosphate could increase the expression of dentin matrix protein-1 and dentin sialoprotein, but downregulate the expression of matrix extracelular phosphoglycoprotein at 7, 14, 21 days. At 28 days of culture, dental pulp stem cels were al successfuly mineralized detected by alizarin red S. There were some red mineralized nodules. These findings indicate that the 5 mmol/L β-glycerophosphate can induce the differentiation of dental pulp stem cels into odontoblasts successfuly, up-regulate the mRNA expression of dentin sialoprotein and dentin matrix protein-1, and meanwhile down-regulate the mRNA expression of matrix extracelular phosphoglycoprotein.

Objective:To investigate the feasibility of tube potential of 80 kV combined with personalized contrast agent protocol in carotid artery CT angiography (CTA).Methods:A total of 136 consecutive patients undergoing neck CTA were prospectively enrolled in this study. The patients were randomly divided into Groups A, B, C and D. Tube potential of 100 kV and 15 s contrast agent injection protocol was used for Group A (53 cases) as conventional group, while tube voltage of 80 kV and 10 s contrast agent injection protocol was used for Groups B, C and D as experimental groups, with the contrast agent dosages of 20, 25 and 30 ml used according to the body weights of ≤50 kg(Group B, 20 cases), 50-70 kg (Group C, 38 cases), and 70-90 kg (Group D, 25 cases), respectively. The subjective and objective evaluation results of image quality and the effective doses were compared among the four groups.Results:The effective doses in Groups B, C and D were 1.54±0.91, 1.89±1.08 and 2.14±1.27 mSv, respectively, significantly lower than that in Group A [(5.66±0.56) mSv] ( F=169.34, P<0.05). The image quality of four groups met the requirements of clinical diagnosis. No significant differences were found in subjective evaluation and diagnostic efficacy of the four groups ( P>0.05). The CT number of carotid artery, signal-to-noise ratio and contrast-to-noise ratio of the neck region were significantly lower in Groups B, C and D compared with Group A ( F=14.9, 12.94, 14.43, P<0.05). The CT numbers of target carotid vessel were all higher than 250 HU. Conclusions:The scanning protocol of low tube potential (80 kV) combined with 10 s contrast agent injection protocol could not only reduce the doses of radiation and contrast agent, but also preserve the diagnosis effect. Thus, this scanning protocol was feasible and valuable in clinical application.

Objective To investigate the accuracy and reproducibility of deep learning algorithms combined with asynchronous calibration quantitative computed tomography(QCT)for measuring bone mineral density(BMD),and to explore the feasibility of using low-dose scanning BMD measurement.Methods European spine phantom(ESP)was scanned with asynchronous calibration QCT and conventional synchronous calibration QCT,respectively,the accuracy and short-term reproducibility was compared.ESP were scanned with asynchronous calibration QCT,matching 120 kVp with five sets of tube currents:20,60,100,140,and 180 mA.Three levels of deep learning image reconstruction(DLIR)and hybrid model-based adaptive statistical iterative reconstruction V(40％ASIR-V)were used for reconstruction.The BMD values of three vertebrae in the ESP were measured.Furthermore,the image noise and contrast-to-noise ratio(CNR)were compared.Results The relative errors(RE)of the three vertebrae of the asynchronous calibration QCT and synchronous calibration QCT were all less than 7％.There was no statistical difference in the BMD values of the two scans at one week interval of the asynchronous calibration QCT(P＞0.05).There were no significant differences in RE among different tube currents or different reconstruction methods(P＞0.05).The image quality of deep learning-based image reconstruction of high strength(DLIR-H)at 20 mA tube current was better than that of 40％ASIR-V at 180 mA,and the radiation dose was reduced by 89％.Conclusion Asynchronous calibration QCT has high accuracy in BMD measurement,and has good repeatability.Asynchronous calibration QCT which combined with DLIR does not affect the accuracy of BMD measurement,and can significantly improve the CNR of images and reduce image noise.

Objective To investigate the effect of lipin1(LPIN1)on the progression of Parkinson's disease(PD)in rats and the possible molecular mechanism of its regulation.Methods The PD rat model was established by injection of 6-Hydroxydopamine hydrobromide(6-OHDA)into the medial forebrain tract of rats,and the LPIN1-overexpressing adenovirus was stably transfected to evaluate the behavioral changes of rats.The content of Fe2+and Glutathione(GSH)and the protein level of tyrosine hydroxylase(TH)in rat brain were detected,and HE staining was used to observe the pathological changes in rat brains.The PD cell model was constructed in vitro,and TH,α-synuclein(α-syn),LPIN1 protein levels and cell viability were detected.LPIN1 small interfering siRNA sequence and overexpression vector and peroxisome proliferator-activated receptor(PPARA)small interfering(siRNA)and overexpression vector were transfected,or ferroptosis inducer erastin was used to treat cell for 24 h,then cells were treated with 6-OHDA for 48h.The levels of Fe2+,reactive oxygen species(ROS),malondialdehyde(MDA),GSH and inflammatory factors in cells were detected to evaluate ferroptosis.Cell viability was detected with CCK-8,and the expressions of ferroptosis related proteins were detected with Western blot.The interacting protein PPARA of LPIN1 was predicted by STRING database and verified by Co-IP analysis.The binding site of PPARA to the promoter of solute carrier family 47 member 1(SLC47A1)was predicted by JASPAR bioinformatics and verified by Ch-IP analysis.Results The fur of the rats in the model group was frightened,and PD symptoms such as continuous tremor,slow movement and weakened activity were shown.The motor behavior and PD symptoms of LPIN1 group were improved/alleviated compared with the model group.Compared with the sham operation group,the total distance of the model group was shortened,the average speed was reduced,and the step length was reduced,while the total resting time was prolonged,the step width was widened,and the gait variation rate was increased,and the differences were significant(t=4.470～26.556,all P＜0.05).Compared with sham operation group,Fe2+content in brain tissue of model group was increased,while GSH content and TH protein expression were decreased,with significance differences(t=8.305,13.305,7.709,all P＜0.05).Compared with the model group,the behavioral evaluation,the level of indexes and the pathological changes of brain tissue in LPIN1 group were improved/alleviated.In addition,6-OHDA decreased PC-12 cell viability,reduced the levels of TH and LPIN1 protein,and increased the level of α-syn protein in a dose-dependent manner,and the differences were significant(F=31.023,7.350,9.124,15.841,all P＜0.05).Silencing LPIN1 intensified the inhibitory effect of 6-OHDA on the viability of PC-12 cells(t=2.209,P＜0.05),and overexpression of LPIN1 could counteract the effect of 6-OHDA.Overexpression of Lpin1 decreased the secretion of IL-1β and IL-6,increased the protein levels of SLC47A1 and GPX4,decreased the levels of Fe2+,MDA and ROS,and increased GSH content(t=3.013～11.639,all P＜0.05).Erastin reversed the inhibitory effect of Lpin1 overexpression on ferroptosis,and reduced the viability of PC-12 cells(t=3.087～7.581,all P＜0.05).LPIN1 interacted with PPARA protein and promoted PPARA expression,while PPARA bound to SLC47A1 promoter and promoted SLC47A1 transcriptional activation.Overexpression of PPARA counteracted the effect of Lpin1 silencing on PC-12 cells.Conclusion Overexpression of LPIN1 may reduce neuronal cell apoptosis by inhibiting ferroptosis mediated by PPARA/SLC47A1 axis,thus alleviating the progression of PD model rats.

Objective:To investigate the molecular genetic and clinical characteristics of MEF2D-BCL9 fusion gene-positive acute B-cell lymphoblastic leukemia (B-ALL), and to provide the reference for the diagnosis and treatment of the disease.Methods:The medical record and experimental examination data of a 18-year-old female MEF2D-BCL9 fusion gene-positive B-ALL patient were retrospectively analyzed. The clinical manifestations and biological characteristics of MEF2D-BCL9 fusion gene-positive B-ALL were summarized.Results:This 18-year-old female patient was treated in a local hospital in December 2018 and was diagnosed as B-ALL. She achieved complete remission after chemotherapy and recurred at 6 months after the initial onset, and then she was admitted to Hebei Yanda Ludaopei Hospital in the 9 months after the initial onset.MEF2D-BCL9 fusion gene was detected through RNA-sequencing (RNA-seq) and verified by using polymerase chain reaction and Sanger sequencing. Bone marrow cell morphology was similar to mature B cells with vacuoles but without characteristic chromosome karyotype abnormalities. The patient achieved remission after VLD regimen chemotherapy, chimeric antigen receptor T-cell (CAR-T) therapy and bridged to allogeneic hematopoietic stem cell transplantation (allo-HSCT). She has maintained complete remission for 2 years at the last follow-up in February 2022.Conclusions:MEF2D-BCL9 fusion gene-positive B-ALL is characterized with high risk, early relapse and poor prognosis. These patients may benefit from CAR-T and allo-HSCT. It further emphasizes the importance of taking MEF2D-BCL9 fusion gene into the detection or identification by using RNA-seq, particularly for those newly diagnosed B-ALL patients in children and adolescents with specific bone marrow morphology.

OBJECTIVE: To detect fusion gene with pathological significance in a patient with refractory and relapsed acute B cell lymphoblastic leukemia (B-ALL) and to explore its laboratory and clinical characteristics. METHODS: Transcriptome sequencing was used to detect potential fusion transcripts. Other laboratory results and clinical data of the patient were also analyzed. RESULTS: The patient was found to harbor TCF3 exon 17-ZNF384 exon 7 in-frame fusion transcript. The minimal residual disease (MRD) has remained positive after multiple chemotherapy protocols including CD19-, CD22- targeted chimeric antigen receptor T cells immunotherapy. The patient eventually achieved complete remission and sustained MRD negativity after allogeneic hemopoietic stem cell transplantation (allo-HSCT). CONCLUSION Transcriptome sequencing can effectively detect potential fusion genes with clinical significance in leukemia. TCF3-ZNF384 positive B-ALL has unique laboratory and clinical characteristics, may not well respond to chemotherapy and immunotherapy, and is more likely to relapse. Timely allo-HSCT treatment may help such patients to achieve long-term disease-free survival. TCF3-ZNF384 positive B-ALL is not uncommon in pediatric patients but has not been effectively identified.
B-Lymphocytes ; Basic Helix-Loop-Helix Transcription Factors/genetics* ; Child ; Hematopoietic Stem Cell Transplantation ; Humans ; Laboratories ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/therapy* ; Trans-Activators/genetics* ; Transcriptome

In recent years, the demand of biologics has increased rapidly. Cell culture process with perfusion mode has become more and more popular due to its high productivity, good quality and high efficiency. In this paper, the unique operation and the details of process optimization for perfusion culture mode are discussed by comparing with traditional batch culture process. Meanwhile, the progress and strategies in the development and optimization of perfusion culture process in recent years are summarized to provide reference for the future development of mammalian cell perfusion culture technology.
Animals ; Batch Cell Culture Techniques ; trends ; Bioreactors ; standards ; CHO Cells ; Cricetulus ; Mammals ; Perfusion

Objective:To investigate the infection spectrum revealed by metagenomics high-throughput next-generation sequencing (mNGS), and to provide a reference for infection diagnosis after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:A total of 64 patients who developed systemic or local infection symptoms after allo-HSCT in Hebei Yanda Lu Daopei Hospital from January 2018 to November 2018 were enrolled. Gene sequences of pathogenic microorganisms in blood, cerebrospinal fluid and bronchoalveolar fluid specimens were detected by using mNGS. The pathogenic microorganisms or suspected pathogens were determined based on the clinical manifestations of patients.Results:There were 97 samples of mNGS detection for 64 patients who underwent allo-HSCT. The most common gram-positive bacteria were staphylococcus haemolyticus (19 times) and staphylococcus (14 times), and the most common gram-negative bacterium was acinetobacter baumannii (8 times). The most common viruses were cytomegalovirus, EB virus and Torque teno virus (35, 22 and 23 times, respectively), and the most common fungi were malassezia globus (14 times) and candida parapsilosis (8 times). There were 3 mycobacterium tuberculosis complexes detected in 3 patients with acute myeloid leukemia who received allo-HSCT. Mycoplasma orale was detected in one patient's sputum, and none parasite was detected.Conclusion:mNGS can comprehensively reveal the infection spectrum of hematologic diseases after allo-HSCT, especially for pathogenic microorganisms that are rare or difficult to cultivate, and it can effectively help the diagnosis of clinically infectious pathogens.