Efficacy and safety of fourth-generation CD19 CAR-T expressing IL7 and CCL19 along with PD-1 monoclonal antibody for relapsed or refractory large B-cell lymphoma / 中华血液学杂志

Objective: This study systematically explore the efficacy and safety of fourth-generation chimeric antigen receptor T-cells (CAR-T), which express interleukin 7 (IL7) and chemokine C-C motif ligand 19 (CCL19) and target CD19, in relapsed or refractory large B-cell lymphoma. Methods: Our center applied autologous 7×19 CAR-T combined with tirelizumab to treat 11 patients with relapsed or refractory large B-cell lymphoma. The efficacy and adverse effects were explored. Results: All 11 enrolled patients completed autologous 7×19 CAR-T preparation and infusion. Nine patients completed the scheduled six sessions of tirolizumab treatment, one completed four sessions, and one completed one session. Furthermore, five cases (45.5%) achieved complete remission, and three cases (27.3%) achieved partial remission with an objective remission rate of 72.7%. Two cases were evaluated for disease progression, and one died two months after reinfusion because of uncontrollable disease. The median follow-up time was 31 (2-34) months, with a median overall survival not achieved and a median progression-free survival of 28 (1-34) months. Two patients with partial remission achieved complete remission at the 9th and 12th months of follow-up. Therefore, the best complete remission rate was 63.6%. Cytokine-release syndrome and immune effector cell-associated neurotoxicity syndrome were controllable, and no immune-related adverse reactions occurred. Conclusion: Autologous 7×19 CAR-T combined with tirelizumab for treating relapsed or refractory large B-cell lymphoma achieved good efficacy with controllable adverse reactions.

Cloning and expression analysis of wpHSP90 gene from Whitmania pigra at different temperatures / 中国中药杂志

HSP90 is a widely distributed molecular chaperone that participates in a variety of cellular processes and plays an important role in the meiosis of germ cells. However, its role in the gonadal development of hermaphroditic Whitmania pigra is not yet clear. To explore the effect of HSP90 on the germ cell development of Wh. Pigra, this study cloned the wpHSP90 gene, performed bioinformatics analysis, and measured its expression levels. The results showed that the cloned wpHSP90 was 2 592 bp in length, with an open reading frame(ORF) of 2 373 bp, encoding 790 amino acids. Prediction analysis revealed 85 phosphorylation modification sites on serine, threonine, and tyrosine residues of the wpHSP90 protein. Structural domain prediction and multiple sequence alignment results showed that wpHSP90 contained two conserved domains of HSP90 and exhibited the highest homology with Helobdella robusta, with a sequence similarity of 80.72%. RT-qPCR results showed that the relative expression level of wpHSP90 in the gonads of 5-month-old Wh. pigra was positively correlated with temperature within the range of 12 ℃ to 28 ℃. The expression level in the female gonads was significantly higher than in the male gonads and correlated with the trend of germ cell development in the ovaries and testes. In conclusion, wpHSP90 may be involved in regulating the development of germ cells, particularly oocytes, in Wh. pigra. This study provides a reference for further research on the gonadal development mechanism in Wh. pigra.

Effects of Wp-PKC on gonadal development of Whitmania pigra / 中国中药杂志

Protein kinase C(PKC) is a kind of kinase which is widely involved in cell proliferation and development. PKC(Wp-PKC) in Whitmania pigra body belongs to classic PKC. In order to investigate the effect of Wp-PKC on the development of Wh. pigra germ cells, 17β-estradiol(17β-E2)(100 ng·mL~(-1)) and methyltestosterone(MT)(150 μg·L~(-1)), 150 μg·L~(-1)(MT)+0.5 mg·L~(-1) PKC, 0.5 mg·L~(-1) PKC inhibitor were added to Wh. pigra culture water, and no addition group(control group) was added, and the effects on the development of Wh. pigra germ cells and the expression of Wp-PKC were observed. The results showed that: Wp-PKC in male gonads was always higher than that in female gonads; MT promoted the development of male gonads in Wh. pigra, while the expression of Wp-PKC was significantly higher than that in the control; 17β-E2 promoted the development of female gonads in Wh. pigra and Wp-PKC expression significantly lower than that of the control; while the development of the female and male gonads in the PKC inhibitor group was inhibited, the expression of Wp-PKC was significantly lower than that of the control. In summary, Wp-PKC may promote the development of Wh. pigra, especially the development of male gonads.

Cloning and expression analysis of protein kinase C gene from Whitmania pigra / 中国中药杂志

Protein kinase C(PKC) is a type of protein kinase widely involved in cell proliferation and development, but the developmental mechanism in the gonads of androgynous animals is still unclear. In order to explore the role of protein kinase C in the development of Whitmania pigra germ cells, the Wh. pigra PKC(Wp-PKC) gene was cloned, bioinformatics analysis was conducted, and fluorescent quantitative PCR was used to analyze the expression of female and male gonads. The results showed that:(1)The cloned Wp-PKC had a full length of 2 580 bp, a relative molecular weight of 76 555.19, and contains an open reading frame encoding 670 amino acids, Wp-PKC was closely related to Danio rerio PKC-α and rat PKC-γ. The similarity of amino acid sequence was 55% and 58%.(2)The protein encoded by Wp-PKC had no signal peptide and was a hydrophilic protein. The secondary structure is mainly composed of random coils, α-helices, extended chains, folds and folds, with the largest proportion of random coils and α-helices. Wp-PKC protein does not contain a transmembrane domain. Multiple sequence alignment and domain prediction analysis show that Wp-PKC contains 4 conserved domains of classical protein kinase C.(3)Fluorescence quantitative results showed that the expression of Wp-PKC in Wh. pigra gonads was positively correlated with the development of germ cells, and the expression in male gonads was significantly higher than that in female gonads. In summary, Wp-PKC is a classic PKC, and Wp-PKC may promote the development of Wh. pigra, especially the development of male gonads, and provide references for further research on the developmental mechanisms of Wh. pigra.

Lipidomics analysis on schisandrol B-induced liver enlargement in mice / 药学学报

Schisandrol B (SolB) is one of the active constituents from a traditional Chinese medicine Schisandra chinensis or Schisandra sphenanthera. Our previous studies found that SolB exerts hepatoprotective effects against drug-induced liver injury and promotes liver regeneration. We further found that SolB significantly induces liver enlargement but the mechanisms remain unclear. The purpose of this study was to investigate the change of lipidome in liver tissues during SolB-induced hepatomegaly. The animal experiment protocol was approved by the Institutional Animal Care and Use Committee at Sun Yat-sen University. Serum and liver samples of male C57BL/6 mice were collected after intraperitoneal injection of SolB (100 mg·kg-1·d-1) for 5 days. Lipidomics analysis was performed using Q Exactive UHPLC-MS/MS system. The results showed that SolB significantly promoted liver enlargement in mice without liver injury and inflammation. Lipid accumulation was observed in the liver tissues after SolB treatment. Thirty-five lipids were identified with significant change and triglycerides (TG) were found to have the most significant increase in SolB-treated group, indicating the increase of energy production during SolB-induced hepatomegaly. This study reveals the impact of SolB on lipid metabolism and provides a potential explanation for liver enlargement induced by SolB.

Effects of MT on growth and gonad development of natural overwintering Whitmania pigra / 中国中药杂志

The objectives of study were to explore the effects of exogenous methyltestosterone( MT) on the growth and gonadal development of overwintering Whitmania pigra. Before overwintering,0. 1,1. 0,10. 0,100. 0,150. 0 μg·L-1 of MT were added to the aquaculture water for 6 weeks. The changes of growth performance,gonad index,endogenous steroid hormones level and internal quality were measured after hibernate for 60 days. Then the tissue slice technique was used to observe the spermary( ovary) of Wh. pigra.The results showed that the body weight,survival rate and gonadal index increased first and then decreased with the increase of exogenous MT concentration; the male gland index was found the highest at the concentration of MT 10. 0 μg·L-1 and the female gland index was the highest at the concentration of MT 1. 0 μg·L-1. The survival rate of Wh. pigra peaked at the concentration of MT 10. 0 μg·L-1.The weight reaches a peak at a concentration of MT 100. 0 μg·L-1( P<0. 05). The number of primary spermatocytes in the testis was negatively correlated with the concentration of exogenous MT. The number of secondary spermatocytes and sperm cells increased first and then decreased. The concentration of secondary spermatocytes was the highest when the concentration of MT was 100. 0 μg·L-1.The number and volume of oocytes in the ovary and the yolk granules increased first and then decreased with the increase of exogenous MT concentration,and the highest was observed at the MT concentration of 100. 0 μg·L-1. The endogenous steroid hormone of Wh.pigra increased first and then decreased with the increase of exogenous MT concentration. The concentration of androgen and progesterone was the highest in MT 100. 0 μg·L-1 treatment( P<0. 05),and the concentration of estrogen was found the highest in MT 10. 0 μg·L-1 treatment( P<0. 05). After adding exogenous MT,Wh. pigra moisture content,acid-insoluble ash content,p H and anti-thrombin activity met the quality criterion of medicinal Wh. pigra in Chinese Pharmacopoeia( 2015 edition). In conclusion,the short-term addition of 1. 0-100. 0 μg·L-1 exogenous MT before hibernation can promote the growth,the development of sperm cells and the antithrombin activity of Wh. pigra.

Establishment of a new genus of filovirus (Mengla virus) entry evaluating system and entry inhibitors discovery / 药学学报

Mengla virus (MLAV), isolated from the bats in China, was identified as a new genus of filovirus in 2019, i.e. Dianlovirus genus of Filoviridae family. Among filoviruses, Ebola virus (EBOV) and Marburg virus (MARV) are the most contagious viruses with mortality rates of 24%-90%. Phylogenetic analysis showed that MLAV was closely related to MARV among the members of filovirus family. MLAV enters into host cells via viral glycoprotein (GP). The recombinant virus study indicated that MLAV has a potential for bat-to-human cross-species transmission. In this study, a GP-mediated MLAV entry evaluating model was established, and by using this model, we investigated the susceptibility of MLAV to the human cell lines sourced from different tissues and the African green monkey kidney cell lines. Four compounds, chloroquine, tetrandrine, clomiphene, and toremifene, which were known as EBOV and MARV entry blockers, were tested for HIV/MLAV-GP infection. It was found that chloroquine effectively blocked the entry of MLAV with the half maximal effective concentration (EC50) of 1.56 μmol·L-1, resembling its anti-EBOV and -MARV activities. To the best of our acknowledge, there is no anti-MLAV drug reported by far, and the identification of chloroquine as an MLAV entry inhibitor may provide an insight for developing anti-filovirus agents.

Chemical constituent analysis for seeds of Celosia argentea by UPLC-ESI-Q-TOF-MS / 中国中药杂志

This Paper aimed to analyze and identify the chemical constituents from the seeds of Celosia argentea by UPLC-ESI-Q-TOF-MS. The analysis was performed on an ACQUITY HSS T3 reverse phase column(2.1 mm ×100 mm, 1.8 μm). The mobile phase consisting of 0.1% formic acid acetonitrile and 0.1% aqueous formic acid was used for gradient elution, and the flow rate was 0.4 mL·min~(-1). Mass spectrometry was applied for the qualitative analysis under positive and negative ionization modes and ESI ion source. Data was analyzed by Masslynx 4.1 software, literatures in SciFinder database, and standards. A total of 49 compounds, including 14 triterpenoids, 17 flavonoids, 11 cyclic peptides, 2 phenols, 2 organic acids, and 3 steroids were putatively identified. Among them, 19 compounds were firstly reported from this species. In-depth chemical constituent analysis for the seeds of C. argentea were accomplished here, and the findings could lay a good foundation for its quality control and clarifying the material basis of its efficacy.

Effects of 17β-E2 on growth and gonad development of overwintering Whitmania pigra / 中国中药杂志

The present study was conducted to explores the effects of short-term addition of 17β-E2 on the growth,gonad development and internal quality of overwintering Whitmania pigra. Before overwintering,0. 0,1. 0,10. 0,25. 0,50. 0,100. 0 μg·L~(-1) of 17β-E2 were added to the aquaculture water for 6 weeks and then hibernated for 60 days. The changes of growth performance,gonad index,morphological structure of spermary( ovary),endogenous steroid hormones level and internal quality were measured. The results showed that the body weight,weight gain rate,specific growth rate,female gonad index,oocyte development and endogenous estrogen level of the leech increased first and then decreased with the increase of the concentration of exogenous 17β-E2,which were higher than those of the control group. The body weight,weight gain rate and specific growth rate of the leech at the concentration of 25 μg·L~(-1)17β-E2 were significantly higher than those of the other groups( P<0. 05),oocyte development and endogenous estrogen levels were significantly higher than those of other groups at the concentration of 50 μg·L~(-1)( P<0. 05). When the concentration of exogenous 17β-E2 was higher than 50 μg·L~(-1),the levels of male gonad index,spermatocyte development,endogenous androgen and progesterone were significantly inhibited( P< 0. 05). There was no significant difference in endogenous corticosteroid levels among the groups. In conclusion,short-term addition of exogenous 17β-E2 of 10-25 μg·L~(-1) could promote the growth of overwintering leeches,oocyte development and antithrombin activity without inhibiting the development of male gonads.

Correlation analysis between cold and heat property of traditional Chinese medicines and light conditions of their original plants in habitat / 中国中药杂志

The current study was conducted to explore the relationship between property of traditional Chinese medicines (TCM) and the light conditions of their original plant in habitats and advance a "light-cold and heat property" hypothesis under the guidance of theory of TCM. In the present study, we selected 127 common Chinese herbs. Association rules mining method was used to find out the association relationship between the cold-heat properties of different TCM and the habitats of their original plant, while using statistical methods to analyze the correlation between light condition and cold-heat property of TCM based on the description of the habitats in the Flora of China and investigation. The results showed that the habitats associated to TCM with cold property are in the order of grassland>curb>hillside>wasteland>gully>farmland>riverside. The habitats that associated to TCM with heat property are: undergrowth>shrub>gully>jungle. For 68 TCMs with cold property, 81.2% of their original plants are light-demanding, 8.7% are shade-demanding, and 8.7% of them showed no obvious light preference. For 59 TCMs with heat property, 62.7% of their original plants are shade-demanding, 20.3% are shade-demanding, and 17.0% of them showed no obvious light preference. Meanwhile, Chi-square test showed that there was a significant difference between the cold-heat property and light preference (<0.01). In summary, there is a significant correlation between the light preference of TCM original plants and their cold-heat properties. In order to ensure the pharmacological effect of TCM, the relationship between the light conditions and the properties of TCM should be considered in the cultivation of medicinal plants for TCM.

Effect of Astragali Radix on growth, immunity and related gene expression of Whitmania pigra / 中国中药杂志

To explore the effect and mechanism of Astragali Radix on growth and immunity of Whitmania pigra, 0, 0.01%, 0.03%, 0.05%, 0.07%, 0.09% of Astragali Radix were added to the daily feeding of Wh. pigra. After 60 days of feeding, the growth performance, activities of digestive enzyme and anti-reverse enzyme, inner quality, the expression levels of GH, IGF-1 and digestive enzyme-related genes were measured. Meanwhile, the effects of heat stress on the living conditions of Wh. pigra were observed and counted, and the expression levels of HSP70 and immune related genes were measured. The results showed that the final weight, weight gain rate, specific growth rate, activities of digestive enzyme and anti-reverse enzyme, the expression levels of GH, IGF-1 and digestive enzyme-related genes in the Astragali Radix group were higher than those in the control group, and with the increase of Astragali Radix concentration, the above-mentioned indexes increased initially and then decreased, and significantly higher in the 0.05% of Astragali Radix group than in the other groups (<0.05). There was no significant difference in the inner quality of Wh. pigra between the Astragali Radix and control groups. The survival rate of Wh. pigra was negatively correlated with heat stress treatment duration. With the prolongation of heat stress treatment duration, the expression levels of HSP70 and immune related genes were increased first and then decreased, and peaked at 24 h. The survival rate and the expression levels of HSP70 and immune related genes in the Astragali Radix group were higher than those in the control group, and was significantly higher in the 0.05% of Astragali Radix group than in the other groups (<0.05). In conclusion, Astragali Radix can increase the activities of digestive enzyme and anti-reverse enzyme, the expression levels of related genes, growth performance, and immunity to heat stress of Wh. pigra. It is suggested to add 0.05% of Astragali Radix in the actual production of Wh. pigra to improve the production profit.

Effects of Cu²⁺ enrichment and release on growth， internal quality and relative gene expression of Whitmania pigra / 中国中药杂志

The present study explored the effects of Cu²⁺enrichment and release on the growth and internal quality of Whitmania pigra, and the regularity of enrichment and release in different tissues of Wh. pigra. In the range of safe concentration(SC), a certain concentration of Cu²⁺ was added to the Wh. pigra for 50 days, and stopped adding for another 50 days. The growth index of Wh. pigra, Cu²⁺ content in different tissues, and the internal quality were determined. The results showed that the average body weight of Wh. pigra in 0.014 mg·L⁻¹ of Cu²⁺ group was significantly higher than that in control group during the experimental period (<0.05), and the mortality rate was lower than that in the control group. The specific growth rate, digestive enzyme activity, growth and digestive enzyme related gene expression of Wh. pigra in Cu²⁺ group were significantly higher than that in the control group (<0.05) during the early 40 days of enrichment, and there was no significant difference in the release period. In Cu²⁺ group, the expression of immune enzyme and immune enzyme related gene of Wh. pigra increased first and then decreased compared with control group (<0.05). The ability of enriched Cu²⁺ in different tissues of Wh. pigra was digestive tract>muscle>skin. The ability of release Cu²⁺ was muscle>digestive tract>skin. There was no significant difference in the internal quality and hirudin gene expression between Cu²⁺ group and control group. In conclusion, Cu²⁺ can improve the expression of digestive enzymes, immune enzyme and related genes of Wh. pigra, promote the growth of Wh. pigra and enhance their immunity, but it doesn't affect the internal quality of Wh. pigra. The Wh. pigra can completely release Cu²⁺ within 30 days.

Preliminary study on changes of sex steroid hormones in periodic development of Whitmania pigra gonad / 中国中药杂志

The present study was conducted to investigate the rule of sex steroid hormones dynamic in the periodic development of Whitmania pigra gonad. The dynamic of sex steroid hormones in different age of Wh. pigra were detected by enzymolysis, extraction and UPLC-MS/MS. The results showed that the concentrations of estrone, estriol, testosterone and progesterone in the Wh. pigra showed M-type curve and peaked in 6-month-old Wh. pigra. The concentrations of above-mentioned four steroid hormones were the lowest in 9-month-old and then increased slightly. The another peaks of testosterone and progesterone were found in 2-month-old and the another peaks of estrone and estriol were obtained in 8-month-old. The concentrations of testosterone and progesterone are slightly higher than estriol during 1-month-old to 4-month-old, and thereafter strone and estriol showed higher concentration than testosterone and progesterone. In summary, the concentrations of four sex steroid hormones in Wh. pigra increased gradually with the maturation of gonads and decreased rapidly after the discharge period, which indicated that the ratio between estrogen, androgen and progesterone may be greatly related to the specificity of gonad development.

Comparative study of ambulatory blood pressure between renovascular hypertensive and essential hypertensive patients / 中华心血管病杂志

<p><b>OBJECTIVE</b>To compare 24 h ambulatory blood pressure changes between patients with renovascular hypertension and essential hypertension.</p><p><b>METHODS</b>The 24 h ambulatory blood pressure of patients with age and gender matched renovascular hypertension (RVH, n = 51) was compared with that of patients with essential hypertension (EH, n = 51).</p><p><b>RESULTS</b>The 24 h, daytime and nighttime systolic blood pressures (SBP), diastolic blood pressures (DBP) and pulse pressures (PP) in RVH were significantly higher than in EH (all P < 0.05), especially the nocturnal SBP (P < 0.05). The SBP and DBP loads in RVH were 58.96% and 35.98% respectively, while blood pressure loads were around 20.00% in EH (P < 0.05). In patients with RVH, The nocturnal blood pressure fall was 5.39%, and only 27.50% patients were dippers, while the nocturnal blood pressure fall was 10.36% and 60.8% patients were dippers in EH.</p><p><b>CONCLUSION</b>RVH patients have higher dynamic BP, PP, BP loads and blunted diurnal rhythm compared to those with EH.</p>

A two-step protocol for induction of rat adipose tissue-derived stem cells into neuron-like cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the differentiation potential of rat adipose tissue-derived cells (ADSCs) into neuron-like cells in vitro using a two-step induction protocol.</p><p><b>METHODS</b>ADSCs isolated from the epididymal fat pads in male SD rats by means of differential attachment were cultured in vitro and subjected to adipogenic induction. After flow cytometric identification of the cell surface antigens CD106, CD11b, CD45, CD49d, CD90 and CD29, the third-passage ADSCs were induced to transdifferentiate into neural stem cell (NSC)-like cells in DMEM/F12 medium containing 10 ng/ml basic fibroblast growth factor (bFGF), 20 ng/ml epidermal growth factor (EGF) and 2% B27. The resultant NSC-like cells were then induced to differentiate into neuron-like cells in the neurobasal medium containing 10 ng/ml brain-derived neurotrophic factor (BDNF), 10 ng/ml glial cell line-derived neurotrophic factor (GDNF) and 1 µmol/L retinoic acid (RA). Immunocytochemistry was employed to identify the expression of the cell surface markers nestin, MAP2 and NeuN.</p><p><b>RESULTS</b>The isolated ADSCs were positive for CD90 and CD29, and oil red O staining of the induced adipose-like cells yielded positive results. The third-passage ADSCs induced for 7 days aggregated as floating cell spheres positive for NSC surface antigen nestin. Further induction in neurobasal medium for 4 h resulted in adhesion of the cell spheres and the formation of cell processes extending from some peripheral cells, suggesting a morphological resemblance to neurons. Most of the cells showed positivity for MAP2 and NeuN.</p><p><b>CONCLUSION</b>ADSCs can be induced to differentiate into neuron-like cells in vitro under appropriate conditions.</p>

The development and challenge of vision prosthesis / 中国医疗器械杂志

This paper introduces the current development and challenges of vision prosthesis.

Recognition of Gaussian pixelate Chinese character based on simulated prosthetic vision / 中国医疗器械杂志

In this paper, Gaussian pixelate Chinese character processing software is designed, and HMD is used to realize the recognition experiment for pixelate Chinese characters based on simulated prosthetic vision. The structure of recognition system, software design and the experiment for determining Gaussian width (sigma) are presented. It is shown that when sigma is 0.235, the recognition program is the best.