Establishment and validation of bioactivity measurement method for recombinant human midkine / 药学学报

To establish and optimize a method for the detection of recombinant human midkine (rhMK) activity and verify its methodology, cell counting kit-8 (cck-8) method was used to measure the proliferation activity of rat knee chondrocytes. The specificity, accuracy, precision, linearity and robustness of the method were also verified in this study. The established method was proven to have good specificity because the buffer of rhMK and recombinant human interleukin-1 receptor antagonist have no obvious active effect; the recoveries of the samples with relative activities of 50%, 75%, 100%, 125%, 150% were in the range of 80.0% to 124.0% by statistical analysis, the relative standard deviations (RSD) of relative potency were all within 20%, the linear correlation coefficient, R2 ≥ 0.98, suggesting that the accuracy, precision and linearity of the method were good; the robustness correlation coefficient, R2 ≥ 0.92 and the ratio of maximum to minimum of sigmoidal dose-response were no less than 1.5, indicating that robustness of the methods was good. In conclusion, a bioactivity measurement method for rhMK was established and fully validated in this study and it provides a reliable method for the bioactivity analysis of rhMK routine samples during the development. This study was approved by the Animal Care and Use Committee of Shanghai Model Organisms Center, Inc. (approval number: 2019-0008-06).

A five-year follow-up report on lung imaging and exercise endurance of a patient with paraquat poisoning / 中华劳动卫生职业病杂志

A patient with paraquat poisoning was followed up for five years, and it was showed that the interstitial lesion areas in chest CT of this patient gradually decreased after acute period of the poisoning and no significant changes were found six months later. After that the density of the lesions gradually reduced, while the cystic air cavities slowly increased. In addition, the patient's exercise endurance gradually improved over time, and the lung function was close to the normal level five years after poisoning. The follow-up report helps clinicians to have a deeper understanding of the long-term outcome of paraquat poisoning.

Genotype-phenotype landscape of pituitary adrenocorticotroph hormone adenoma / 中华内分泌代谢杂志

Objective:Cushing′s disease(CD) is caused by the pituitary adrenocorticotroph hormone(ACTH) secreting adenomas, leading to increased serum cortisol levels and various abnormal metabolic processes. Untreated CD is linked to high mortality, thus it is critical to elucidate its pathogenesis. This study aims to explore the pathogenesis of pituitary ACTH adenomas using whole-genome sequencing analysis.Methods:Fresh tumor tissues and peripheral blood samples were collected in 9 confirmed cases of pituitary ACTH adenomas who underwent surgery. Whole genome sequencing was then performed, followed by analysis and verification of single nucleotide mutations, copy number variation(CNV) and chromosome structure variations.Results:Somatic USP8 mutations(p.Ser718del, p. Ser718Pro, p. Pro720Arg, p. Pro720Gln) were found in 5 patients, with a rate of 55.6%; CNV of USP8 was detected in 1 patient; TP53(p.Cys135Tyr), NF1(p.Val1049Glufs*11) and KMT2C(c.3323+ 1G>A) mutations were identified in 1 patient harboring wild-type USP8. CNV analysis showed a loss of heterozygosity in multiple chromosomes in a wild-type USP8 patient. Structural variations were found in 2 with unknown significance. No germline gene mutations were detected in this study.Conclusion:Somatic USP8 mutations, increased copy number of USP8, variations of tumor-related genes such as TP53 and extensive somatic CNV all contribute to pathogenesis of CD. Chromosomal structure variations may suggest high-risk pituitary ACTH adenomas, and call for frequent follow-up and aggressive treatment.

Automatic diagnosis and assessment of bone metastases on bone scans based on deep learning / 中华核医学与分子影像杂志

Objective:To develop an approach for the automatic diagnosis of bone metastasis and to design a parameter of quantitative evaluation for tumor burden on bone scans based on deep learning technology.Methods:A total of 621 cases (389 males, 232 females, age: 12-93 years) of bone scan images from the Department of Nuclear Medicine in Tenth People′s Hospital of Tongji University from March 2018 to July 2019 were retrospectively analyzed. Images were divided into bone metastasis group and non-bone metastasis group. Eighty percent of the cases were randomly extracted from both groups as the training set, and the rest of cases were used as the test set. A deep residual convolutional neural network ResNet34 was used to construct the classification model and the segmentation model. The sensitivity, specificity and accuracy were calculated and the performance differences of the classification model in different age groups (15 cases of <50 years, 75 cases of ≥50 and <70 years, 33 cases of ≥70 years) were analyzed. The regions of metastatic bone lesions were automatically segmented by the segmentation model. The Dice coefficient was used to evaluate the effect of the segmentation model and the manual labeled results. Finally, the bone scans tumor burden index (BSTBI) was calculated to assess the tumor burden of bone metastases.Results:There were 280 cases with bone metastases and 341 cases with non-bone metastases, including 498 in training set and 123 in test set. The classification model could accurately identify bone metastases, with the sensitivity, specificity and accuracy of 92.59%(50/54), 85.51%(59/69) and 88.62%(109/123), respectively, and it performed best in the <50 years group (sensitivity, 2/2; specificity, 12/13; accuracy, 14/15). The specificity in the ≥70 years group (8/12) was the lowest. The Dice coefficient of bone metastatic area and bladder area were 0.739 and 0.925 in the segmentation model, which performed similarly in the three age groups. Preliminary results showed that the value of BSTBI increased with the increase of the number of bone metastatic lesions and the degree of 99Tc m-MDP uptake. The machine learning model in this study took (0.48±0.07) s for the entire analysis process from input to the final BSTBI calculation. Conclusions:The deep learning based on automatic diagnosis framework for bone metastases can automatically and accurately identify segment bone metastases and calculate tumor burden. It provides a new way for the interpretation of bone scans. The proposed BSTBI may be used as a quantitative evaluation indicator in the future to assess the tumor burden of bone metastases based on bone scans.

The research progress on the structure pharmaceutics and dosage form structures of traditional Chinese medicines / 药学学报

Traditional Chinese medicine (TCM) preparations have made tremendous progresses in modernization, whereas there exist relatively few researches pertaining to preparation structures. As demonstrated by the theory and practice of structure pharmaceutics, the structure properties of dosage forms have significant influences on the quality and efficacy of drugs, which might offer reference for the research and development of TCM dosage forms. With the application of synchrotron radiation X-ray micro-computed tomography (SR-μCT) and other novel technologies in recent years, researches in structure pharmaceutics have made huge advancement, which provide reference and methodology basis for the study of TCM preparations. The article generalized and summarized the recent progresses and methods in the structure researches of pharmaceutics and TCM preparations, and further explored the significance of the researches of structure of TCM preparations. It is expected to provide the basis for the dosage form design, production process improvement, and quality evaluation of TCM and promote the modernization of TCM preparations.

Effect and Involved Mechanism of RSL3-induced Ferroptosis in Acute Leukemia Cells MOLM13 and Drug-resistant Cell Lines / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect and involved mechanism of RSL3 on ferroptosis action in acute leukemia cells MOLM13 and its drug-resistant cells.@*METHODS@#After MOLM13 treated with RSL3, CCK-8 assay was performed to detect cell viability, flow cytometry was used to detect the reactive oxygen species (ROS) level of the cells, RT-qPCR and Western blot were used to detect the expression of glutathione peroxidase 4 (GPX4). After MOLM13/IDA and MOLM13/Ara-C, the drug-resistant cell lines were constructed, the ferroptosis induced by RSL3 was observed. Bone marrow samples were collected from patients with acute monocytic leukemia. RT-qPCR and Western blot were performed to detect the expression of related genes and proteins involved in ferroptosis pathway.@*RESULTS@#RSL3 significantly inhibited the cell viability of MOLM13 and increased the intracellular ROS level, which were partially reversed by ferrostatin-1. The mRNA and protein expression of GPX4 decreased in MOLM13 treated with RSL3. RSL3 inhibited the viability of MOLM13/IDA and MOLM13/Ara-C cells more strongly than that of non-drug resistant cells, also increased the intracellular ROS level . The cytotoxic effects were partially reversed by ferrostatin-1. The mRNA and protein expressions of GPX4 in MOLM13/IDA and MOLM13/Ara-C cells were higher than those in non-drug resistant cells. The mRNA and protein levels of GPX4 in bone marrow of relapsed/refractory acute mononuclear leukemia patients were higher than those of ordinary acute mononuclear leukemia patients.@*CONCLUSION@#RSL3 can induce non-drug resistant cells MOLM13 ferroptosis by inhibiting GPX4 activity. MOLM13/IDA and MOLM13/Ara-C are more sensitive to RSL3 compared with non-drug resistant cells MOLM13, which may be caused by the differences in GPX4 expression. The expressions of GPX4 mRNA and protein in relapsed/refractory acute mononuclear leukemia are higher than those in ordinary acute mononuclear leukemia.

Arthroscopic Bankart repair combined with Reimplissage for the treatment of recurrent anterior dislocation of shoulder in adults with Hill-Sachs defect / 中国骨伤

OBJECTIVE@#To investigate the method and clinical effects of the treatment of recurrent anterior dislocation of shoulder with Hill-Sachs injury by arthroscopic Bankart repair and Remplissage.@*METHODS@#From March 2016 to March 2019, 106 patients with recurrent anterior dislocation of shoulder with glenoid bone defect less than 20% underwent arthroscopic Bankart repair, including 76 males and 30 females, aged from 18 to 45 (27.3±8.6) years, 59 cases of left shoulder and 47 cases of right shoulder. Range of motion (ROM), American Shoulder and Elbow Surgeons(ASES) score, Constant-Murley score and Rowe score were used to evaluate shoulder functionand stability before and after operation.@*RESULTS@#All patients were followed up, and the duration ranged from 21 to 60 months, with a mean of (41.5± 8.5) months. One patient developed infection after operation, and the infection was controlled after arthroscopic debridement again. The remaining patients did not have clinical complications such as infection, intra articular hematocele and redislocation. Shoulder flexion and lifting increased from (158.33±15.72) ° preoperatively to (169.43±10.04) ° at the latest follow up, and internal rotation changed from T7 (T4 to T10) preoperatively to T8 (T5 to T10) at the latest follow up;the average lateral external rotation and abduction 90 ° external rotation decreased from (58.46±15.51) ° preoperatively and (99.37±14.09) ° to (53.18±14.90) ° and (92.52±13.10) ° at the latest follow up, respectively. The ASES score, Constant -Murley score and Rowe score were significantly improved.@*CONCLUSION@#The clinical effect of rehabilitation of Bankart repair combined with Remplissageunder arthroscopy in the treatment of recurrent dislocation of shoulder joint in adults with Hill-Sachs defect is satisfactory. Although the external rotation function is weaker than that before operation, it can effectively reconstruct the shoulder function and avoid the occurrence ofdislocation after operation.

Projects status analysis of occupational respiratory diseases funded by the National Natural Science Foundation of China from 1987 to 2018 / 中国职业医学

OBJECTIVE: To analyze the funding status of the National Natural Science Fund(NNSF) in the field of occupational respiratory disease(ORD) research in China. METHODS: Through the NNSF Committee Science Foundation sharing service network database, we collected the information of funded projects in the field of ORD research in China from 1987 to 2018, and analyzed the funding expenditure, funding categories, supporting units and research contents. RESULTS: A total of 106 NNSF funded projects were obtained in the research field of ORD in China in the period 1987-2018, and the total amount of funding was 37.945 million yuan. The number of funded projects, the average and the total amounts of fundings showed an increasing trend year by year(all P<0.05). The main types of funded projects were general projects and Youth Science Foundation projects(97.2%). The funded projects and funds were mainly concentrated in colleges and universities, accounting for 79.2% and 83.4% respectively. Beijing(29 items, 27.4%), Hebei(13 items, 12.3%) and Jiangsu(11 items, 10.4%) were the top three provinces and municipalities in the number of funded projects. Silicosis, pulmonary fibrosis, quartz, alveolar macrophages and acute lung injury were the top five keywords of funded project items, and the research types mainly focused on the mechanism of diseases. CONCLUSION: The NNSF funding is imbalanced in the types of funding, supporting units, and regional distribution on ORD research field in China. It is suggested that the funding structure of NNSF should be optimized, interdisciplinary research should be encouraged and expanded, and cooperation among different institutions and regions should be promoted.

Seroprevalence of IgM and IgG Antibodies against SARS-CoV-2 in Asymptomatic People in Wuhan: Data from a General Hospital Near South China Seafood Wholesale Market during March to April in 2020 / 生物医学与环境科学（英文）

The aim of this study was to estimate the seroprevalence of immunoglobulin M (IgM) and G (IgG) antibodies against SARS-CoV-2 in asymptomatic people in Wuhan. This was a cross-sectional study, which enrolled 18,712 asymptomatic participants from 154 work units in Wuhan. Pearson Chi-square test,

Risk factors for metabolic bone disease of prematurity in very/extremely low birth weight infants: a multicenter investigation in China / 中国当代儿科杂志

OBJECTIVE@#To investigate the incidence rate and risk factors for metabolic bone disease of prematurity (MBDP) in very low birth weight/extremely low birth weight (VLBW/ELBW) infants.@*METHODS@#The medical data of 61 786 neonates from multiple centers of China between September 1, 2013 and August 31, 2016 were retrospectively investigated, including 504 VLBW/ELBW preterm infants who met the inclusion criteria. Among the 504 infants, 108 infants diagnosed with MBDP were enrolled as the MBDP group and the remaining 396 infants were enrolled as the non-MBDP group. The two groups were compared in terms of general information of mothers and preterm infants, major diseases during hospitalization, nutritional support strategies, and other treatment conditions. The multivariate logistic regression analysis was used to investigate the risk factors for MBDP.@*RESULTS@#The incidence rate of MBDP was 19.4% (88/452) in VLBW preterm infants and 38.5% (20/52) in ELBW preterm infants. The incidence rate of MBDP was 21.7% in preterm infants with a gestational age of < 32 weeks and 45.5% in those with a gestational age of < 28 weeks. The univariate analysis showed that compared with the non-MBDP group, the MBDP group had significantly lower gestational age and birth weight, a significantly longer length of hospital stay, and a significantly higher incidence rate of extrauterine growth retardation (@*CONCLUSIONS@#A lower gestational age, hypocalcemia, extrauterine growth retardation at discharge, and neonatal sepsis may be associated an increased risk of MBDP in VLBW/ELBW preterm infants. It is necessary to strengthen perinatal healthcare, avoid premature delivery, improve the awareness of the prevention and treatment of MBDP among neonatal pediatricians, and adopt positive and reasonable nutrition strategies and comprehensive management measures for preterm infants.

Effect of Electro-acupuncture on Expression of IRS-1/PI3K/GLUT4 Pathway in Ovarian Granulosa Cells of Infertile Patients with Polycystic Ovary Syndrome-Insulin Resistance of Phlegm-Dampness Syndrome / 中国结合医学杂志

OBJECTIVE@#To evaluate the effect of electro-acupuncture (EA) in infertile patients with phlegm-dampness polycystic ovary syndrome-insulin resistance (PCOS-IR).@*METHODS@#Seventy-six PCOS-IR patients who underwnet in vitro fertilization and embryo transfer (IVF-ET) were equally assigned to two groups according to a random digital table: the EA group and the control group, with 38 cases in each group. Before undergoing IVF, the two groups were treated with EA or pseudo-acupuncture, respectively, for 3 menstrual cycles. The intervention was 25 min twice a week until the day of oocyte collection. The selected acupoints were Zhongwan (RN 12), Tianshu (ST 25), Daheng (SP 15), Daimai (GB 26), Qihai (CV 6), Guanyuan (CV 4), and bilateral points including Xuehai (SP 10), Fenglong (ST 40), Zusanli (ST 36), and Yinlingquan (SP 9). Evaluation of phlegm-dampness syndrome score and IR score were carried out before and after treatment. Additionally, the number of oocytes retrieved, transplantable embryo rate, high-quality embryo rate, clinical pregnancy rate and live birth rate were compared between the two groups. Real-time polymerase chain reaction analysis was used to monitor the mRNA expression of the insulin receptor substrate (IRS-1), phosphatidylinositiol 3-kinase (PI3K) and glucose transport factor 4 (GLUT4) in ovarian granulosa cells.@*RESULTS@#EA treatment reduced the phlegm-dampness syndrome score as well as the IR scores compared with the control group (P0.05). Moreover, the transplantable embryo rate [49.0% (284/580) vs. 41.9% (273/652)], high-quality embryo rate [36.6% (104/284) vs. 27.8% (76/273)], and live birth rate [50% (19/38) vs. 26.3% (10/38)] in the EA group were significantly higher than in the control group (P<0.05). Gene expression analyses revealed significantly elevated IRS-1, PI3K and GLUT4 mRNA in ovarian granulosa cells of the EA group compared with the control group (P<0.05).@*CONCLUSIONS@#EA may ameliorate the effects of phlegm-dampness syndrome and ovarian IR in PCOS-IR patients. Mechanistically, this effect might be through an upregulation of the IRS-1/PI3K/GLUT4 signaling pathway, which may result in improved oocyte quality and embryonic development potential. (Registration No. ChiCTR1800015453).

Three new secoiridoid glycosides from the flower buds of Lonicera japonica / 中国天然药物

Three new secoiridoid glycosides, named lonijapoglycol A (1), aldosecolohanin C (2) and aldosecolohanin B (3), together with three known ones (4-6), have been isolated from the flower the buds of Lonicera japonica. All the structures were identified by spectroscopic analyses. Lonijapoglycol A (1) expressed significant anti-inflammatory activity to inhibit the release of β-glu-curonidase induced by platelet-activating factor in rat polymorphonuclear leukocytes with an IC value of 3.76 μmol·L.

Three butylphthalide derivatives from the Rhizome of Ligusticum chuanxiong / 药学学报

Three butylphthalide derivatives were isolated from the Rhizome of Ligusticum chuanxiong using a series of isolation and purification approaches including macroporous resin, ODS-A column, Sephadex LH-20 and preparative HPLC. These structures were elucidated based on extensive spectroscopic data (UV, IR, HR-ESI-MS and NMR) and identified as (3Z,3aE)-(6R,7R,2'S)-6-hydroxy-7-(2-carboxyl-2-hydroxyethylthio)-3-(2-hydroxybutylidene)-4,5,6,7-tetrahydro-phthalide (1), (3Z,3aZ)-3-butylidene-6,7-dihydroxy-4,5,6,7-tetrahydro-phthalide 7-O-α-D-glucopyranosyl-(1→2)-β-D-fructo-furanoside (2) and 3-(3-β-D-glucopyranosyloxy-butylidene)-7-hydroxy-phthalide (3).

Mucormycosis in patients with hematological diseases: seven cases reports and literature review / 中华血液学杂志

Objective: To explore the clinical characteristics, diagnosis, treatment and prognosis of patients with hematological diseases that complicated by mucormycosis, and to improve the understanding and clinical diagnosis and treatment of the disease. Methods: The clinical data of 7 patients suffering from mucormycosis during September 2012 and September 2018 were retrospectively analyzed, and their clinical characteristics, treatment process and prognosis were analyzed. Results: Of 7 patients, there were 4 males and 3 females, with a median age of 36 (19-79) years old. Two patients were diagnosed as acute myeloid leukemia as the underlying disease, the other 5 patients suffered from acute B lymphoblastic leukemia, peripheral T cell lymphoma, chronic myelocytic leukemia in blastic phase, myeloproliferative neoplasm and severe aplastic anemia after transplantation, respectively. Among them, disease types of mucormycosis were pulmonary in 4 patients, rhino-orbital-cerebral in 1 patient, cutaneous in 1 patient and disseminated in 1 patient. All the cases were confirmed by biopsy histopathology. The treatment drugs were amphotericin B or liposomal amphotericin B, and posaconazole. Surgical treatment was performed in 4 patients, 3 out of 4 achieved radical debridement, and the other one had local debridement. Two patients were cured, 1 patient was improved and 4 patients died. Conclusions: The clinical manifestation and image feature of mucormycosis in patients with hematological diseases were diverse, and the mortality rate is high, diagnosis mainly depends on histopathology. Early diagnosis, control of underlying disease, improvement of immunosuppressive status, timely effective antifungal therapy and radical surgical debridement are the key points for improving the survival rate of patients with hematological diseases complicated by mucormycosis.

Efficacy and safety of CD19 chimeric antigen receptor T cells for the treatment of 22 patients with B-cell lymphoma / 中华血液学杂志

Objective: To investigate the efficacy and safety of CD19 chimeric antigen receptor T (CAR-T) lymphocytes for the treatment of B cell lymphoma. Methods: A total of 22 patients with B-cell lymphoma from February 1, 2017 to July 1, 2018 were reviewed to evaluate the efficacy and adverse reactions of CD19 CAR-T. Results: Of 22 patients with B-cell lymphoma received CD19 CAR-T cells, the median dose of CAR-T cells was 7.2 (2.0-12.0) ×106/kg. Nine of 12 cases of relapse refractory patients were overall response. Complete remission (CR) occurred in 2 of 12 patients, partial remission (PR) in 7 of 12 patients. The overall response in minor residual disease positive (MRD) group was 8 of 10 patients. CD19 CAR-T cells proliferated in vivo and were detectable in the blood of patients. The peak timepoints of CAR-T cells proliferated in the relapsed refractory and MRD positive groups were 12 (5-19) and 4.5 (1-12) days after treatment respectively, and among peripheral blood cells, CAR-T cells accounted for 10.10% (3.55%-24.74%) and 4.02% (2.23%-28.60%) of T lymphocytes respectively. The MRD positive patients achieved sustained remissions during a median follow-up of 8 months (rang 3-18 months) . None of all the patients relapsed during a median follow-up time of 10 months (3-18 months) . However, 7 PR responders of the relapsed refractory patients maintained a good condition for 1.5-6.0 months. One patient bridged to hematopoietic stem cell transplantation, another one sustained remission for 12 months. Cytokine-release syndrome (CRS) occurred in 14 patients with grade 1-2 CRS in MRD positive group and grade 3 CRS in relapsed refractory group. Conclusions: CAR-T cell therapy not only played a role in the rescue treatment of relapsed and refractory patients, but also produced a surprising effect in the consolidation and maintenance of B-cell lymphoma. CD19 CAR-T cells might be more effective in the treatment of MRD positive B-cell lymphoma patients than in the refractory or relapsed cases. High response rate was observed with fewer adverse reactions.

The usage of comprehensive geriatric assessment in elderly patients with acute myeloid leukemia: a multicenter, prospective study / 中华血液学杂志

Objective: To evaluate the feasibility and potential value of comprehensive geriatric assessment (CGA) in elderly (≥60 years) patients with newly diagnosed acute myeloid leukemia (AML) in China. Methods: The CGA results of 83 newly diagnosed AML (non-APL) patients from 16 hospitals in Beijing and Tianjin between March 2016 and December 2017 were prospectively collected and analyzed. The clinical data, treatment and follow-up information were also collected. Results: Of 83 newly diagnosed elderly AML patients, 81 patients (97.6%) completed all designated CGA assessment. The median number of impaired scales of the CGA assessment in the studied population was 2(0-6). Sixteen patients (19.3%) showed no impairments according to the geriatric assessment scales implem ented by this study. The distributions of impaired scales were as follows: impairment in ADL, 55.4%; IADL impairment, 42.2%; MNA-SF impairment, 48.2%; cognitive impairment, 15.7%; GDS impairment, 31.7%; HCT-CI impairment, 19.5%, respectively. In patients with "good" ECOG (n=46), the proportion of impairment for each CGA scale ranged from 6.5% to 37.0% and 32 patients (68.9%) had at least one impaired CGA scale. Survival analysis showed that the number of impaired scales of the CGA was significantly correlated with median overall survival (P=0.050). Conclusions: CGA was a tool with feasibility for the comprehensive evaluation in elderly AML patients in China. Combined with age and ECOG, CGA may be more comprehensive in assessing patients' physical condition.

A three-dimensional finite element model of mesio-occluso-distal cavity zirconium dioxide inlay and its bonding interface:a stress analysis / 中国组织工程研究

BACKGROUND: Zirconia inlays have good mechanical, biocompatible and aesthetic properties in the field of dental prosthodontics, but there is no consensus on the standards of zirconia inlays for clinical cavity design, dental preparation and selection of bonding materials. OBJECTIVE: To investigate the stress distribution and characteristics of the bonding interface, tooth and periodontal tissues of the zirconia inlay model after 3 M RelyX Unicem or vario-link resin adhesive bonding. METHODS: Micro-CT was used to scan the right mandibular third molars. A three-dimensional finite element model of MOD zirconia inlay with different adhesives (3 M RelyX Unicem resin binder, vario-link resin binder) and cavity types (2, 3, 4 mm in the depth of the joint cavity) was constructed by software Mimics, Goemagic Studio and NX 10.0. Using ANSYS Workbench mesh generation, the stress distribution of each model was analyzed after loading 10 N·mm torque, 45° 175 N, 90° 600 N. RESULTS AND CONCLUSION: (1) After loading 10 N·mm torque, the bonding agent equivalent stress and root surface equivalent stress of the model with 3 mm cavity depth were largest, so the stress on the bonding interface, crown and root was largest. (2) After loading 175 N at 45° lingual direction, the bonding interface of the model with 4 mm cavity depth undertook a higher stress. The stress values of the root and periodontal tissue of 3 M RelyX Unicem resin bonding agent model with the same cavity depth were higher. (3) After loading 600 N at 90°, the root force of the model with 3 mm cavity depth was largest. The crown, root and periodontal tissue of the 3 M RelyX Unicem resin bonding agent model with the same cavity depth undertook a higher stress. The bonding interface of the vario-link resin bonding agent model was under greater stress. (4) The regions of stress concentration areas included the bifurcation zone of the root, the inlay edge line, the roof of pulp chamber, the gingival wall, 1/3 mesial part of the buccal surface near the neck, and 1/3 mesial part of the lingual surface near the neck. (5) All these findings indicate that vario-link resin adhesive and 3 M RelyX Unicem resin adhesive are suitable for the bonding of zirconia inlay, but the vario-link adhesive strength is larger and the bonding interface stress is larger. The factors such as cavity design, residual tooth tissue resistance, retention shape, and periodontal support should be considered comprehensively. The optimization of cavity design, tooth preparation, cushion bottom and high inlay should be adopted, in order to improve the long-term repair effect of zirconia inlay. However, further clinical trials are needed to verify the results of three-dimensional finite element model.

Effect on sperm quality of asthenospermia and oligospermia treated with grain-moxibustion combined with medicine therapy / 中国针灸

OBJECTIVE@#To observe the clinical effect of grain-moxibustion combined with medicine therapy for asthenospermia and oligospermia.@*METHODS@#A tatal of 60 patients were randomized into an observation group (30 cases) and a control group (30 cases) according to 1︰1 ratio. In the control group, vitamin E capsules were taken orally one capsule each time, twice a day, and pills 6 g each time, three times a day for a total of 3 months. In the observation group, grain-moxibustion was applied at Guanyuan (CV 4)，Shenshu (BL 23) and Zusanli (ST 36) based on the control group, once a week for 3 months, with a total of 12 times. The sperm concentration and sperm progressive motility were measured by automatic sperm quality analysis system in the two groups, and the clinical effects were compared. Sperm DNA fragmentation index (DFI) in the observation group was measured by sperm nucleus chromosome structure assay (SCSA).@*RESULTS@#①The sperm concentrations and sperm progressive motilities after 1-month, 2-month and 3-month of treatment were increased compared with those before treatment in the two groups (<0.01), and they were increased with time. In the two groups, 2-month and 1-month of treatment, 3-month and 2-month of treatment were compared, the sperm concentrations and sperm progressive motilities were significantly increased (<0.01). The sperm concentrations after 1-month, 2-month and 3-month of treatment in the observation group were higher than those in the control group (<0.01), the sperm progressive motility after 3-month of treatment in the observation group was higher than that in the control group (<0.05). ②After 3-month of treatment，the DFI in the observation group was significantly reduced compared with that before treatment (<0.01). ③The total effective rate in the observation group after 3-month of treatment was 86.7% (26/30), which was superior to 63.3% (19/30) in the control group (<0.05).@*CONCLUSION@#Grain-moxibustion combined with medicine therapy can improve sperm concentration and sperm progressive motility, enhance the integrity of sperm DNA.

Study of Recellularized Human Acellular Arterial Matrix Repairs Porcine Biliary Segmental Defects

BACKGROUND: With the popularity of laparoscopic cholecystectomy, common bile duct injury has been reported more frequently. There is no perfect method for repairing porcine biliary segmental defects.METHODS: After the decellularization of human arterial blood vessels, the cells were cultured with GFP⁺ (carry green fluorescent protein) porcine bile duct epithelial cells. The growth and proliferation of porcine bile duct epithelial cells on the human acellular arterial matrix (HAAM) were observed by hematoxylin-eosin (HE) staining, electron microscopy, and immunofluorescence. Then, the recellularized human acellular arterial matrix (RHAAM) was used to repair biliary segmental defects in the pig. The feasibility of it was detected by magnetic resonance cholangiopancreatography, liver function and blood routine changes, HE staining, immunofluorescence, real-time quantitative PCR (RT-qPCR), and western blot.RESULTS: After 4 weeks (w) of co-culture of HAAM and GFP? porcine bile duct epithelial cells, GFP⁺ porcine bile duct epithelial cells grew stably, proliferated, and fused on HAAM. Bile was successfully drained into the duodenum without bile leakage or biliary obstruction. Immunofluorescence detection showed that GFP-positive bile duct cells could still be detected after GFP-containing bile duct cells were implanted into the acellular arterial matrix for 8 w. The implanted bile duct cells can successfully resist bile invasion and protect the acellular arterial matrix until the newborn bile duct is formed.CONCLUSION: The RHAAM can be used to repair biliary segmental defects in pigs, which provides a new idea for the clinical treatment of common bile duct injury.

Effect of PD-1 inhibitor Nivolumab on the proliferation and cytotoxicity of anti-CD19 chimeric antigen receptor T cells / 中华血液学杂志

Objective: To Evaluation the effect of PD-1 inhibitor Nivolumab on the proliferation and cytotoxicity of anti-CD19 chimeric antigen receptor T cells (CD19-CAR-T) in vitro. Methods: Five patients with high PD-1 expression in peripheral blood and five healthy volunteers were selected. These peripheral blood mononuclear cells were used as the source of T cells to prepare CD19-CAR-T cells. Different doses (72, 36, 18 μg/ml) of Nivolumab was added on day 8 to the culture medium. Patient T cells incubated with 72 μg/ml Nivolumab and CD19-CAR-T cells of healthy volunteers were used as controls. CCK-8, lactate dehydrogenase (LDH) cytotoxicity assay and ELASA were used to detect the proliferation capacity, the specific cytotoxicity and the inflammatory factor secretion. Results: ①T cells from patients with high expression of PD-1 as the source of CD19-CAR-T cells did not affect transfection rate compared with that of healthy volunteers [(32.80±7.22)% vs (35.10±5.84)%, t=-0.554, P=0.593]. ②Incubation of CD19-CAR-T cells with 72 μg/ml Nivolumab did not affect CD19-CAR-T cell proliferation, but its cytotoxicity was significantly higher than that of CD19-CAR-T cells alone or patients' T cells +72 μg/ml Nivolumab (all P<0.001), there was no significant difference in the killing activity between the 72 μg/ml and 36 μg/ml Nivolumab treated CD19-CAR-T cells on Pfeiffer cells (P=0.281, 0.267, respectively), and they were all higher than those of 18 μg/ml Nivolumab treated CD19-CAR-T cells (all P<0.001). ③Different doses of PD-1 inhibitor Nivolumab combined with CD19-CAR-T cells does not affect the secretion of IFN-γ and IFN-α (all P>0.05). Conclusion: Combination of 36 μg/ml PD-1 inhibitor and CD19-CAR-T cells could reduce the drug toxicity and enhance the cytotoxicity.