Single-cell transcriptomic analysis reveals a novel cell state and switching genes during hepatic stellate cell activation in vitro.

BACKGROUND: The transformation of hepatic stellate cell (HSC) to myofibroblast is a key event during liver fibrogenesis. However, the differentiation trajectory of HSC-to-myofibroblast transition and the switching genes during this process remains not well understood. METHODS: We applied single-cell sequencing data to reconstruct a single-lineage pseudotime trajectory of HSC transdifferentiation in vitro and analyzed the gene expression patterns along the trajectory. GeneSwitches was used to identify the order of critical gene expression and functional events during HSC activation. RESULTS: A novel cell state during HSC activation was revealed and the HSCs belonging to this state may be an important origin of cancer-associated fibroblasts (CAFs). Combining single-cell transcriptomics with GeneSwitches analyses, we identified some distinct switching genes and the order at which these switches take place for the new state of HSC and the classic culture-activated HSC, respectively. Based on the top switching genes, we established a four-gene combination which exhibited highly diagnostic accuracy in predicting advanced liver fibrosis in patients with nonalcoholic fatty liver disease (NAFLD) or hepatitis B (HBV). CONCLUSION: Our study revealed a novel cell state during HSC activation which may be relevant to CAFs, and identified switching genes that may play key roles in HSC transdifferentiation and serve as predictive markers of advanced fibrosis in patients with chronic liver diseases.

Longitudinal characteristics of lymphocyte responses and cytokine profiles in the peripheral blood of SARS-CoV-2 infected patients.

BACKGROUND: The dynamic changes of lymphocyte subsets and cytokines profiles of patients with novel coronavirus disease (COVID-19) and their correlation with the disease severity remain unclear. METHODS: Peripheral blood samples were longitudinally collected from 40 confirmed COVID-19 patients and examined for lymphocyte subsets by flow cytometry and cytokine profiles by specific immunoassays. FINDINGS: Of the 40 COVID-19 patients enrolled, 13 severe cases showed significant and sustained decreases in lymphocyte counts [0·6 (0·6-0·8)] but increases in neutrophil counts [4·7 (3·6-5·8)] than 27 mild cases [1.1 (0·8-1·4); 2·0 (1·5-2·9)]. Further analysis demonstrated significant decreases in the counts of T cells, especially CD8+ T cells, as well as increases in IL-6, IL-10, IL-2 and IFN-γ levels in the peripheral blood in the severe cases compared to those in the mild cases. T cell counts and cytokine levels in severe COVID-19 patients who survived the disease gradually recovered at later time points to levels that were comparable to those of the mild cases. Moreover, the neutrophil-to-lymphocyte ratio (NLR) (AUC=0·93) and neutrophil-to-CD8+ T cell ratio (N8R) (AUC =0·94) were identified as powerful prognostic factors affecting the prognosis for severe COVID-19. INTERPRETATION: The degree of lymphopenia and a proinflammatory cytokine storm is higher in severe COVID-19 patients than in mild cases, and is associated with the disease severity. N8R and NLR may serve as a useful prognostic factor for early identification of severe COVID-19 cases. FUNDING: The National Natural Science Foundation of China, the National Science and Technology Major Project, the Health Commission of Hubei Province, Huazhong University of Science and Technology, and the Medical Faculty of the University of Duisburg-Essen and Stiftung Universitaetsmedizin, Hospital Essen, Germany.

Lack of human-like extracellular sortilin neuropathology in transgenic Alzheimer's disease model mice and macaques.

BACKGROUND: Alzheimer's disease (AD) is a devastating neurodegenerative disorder bearing multiple pathological hallmarks suggestive of complex cellular/molecular interplay during pathogenesis. Transgenic mice and nonhuman primates are used as disease models for mechanistic and translational research into AD; the extent to which these animal models recapitulate AD-type neuropathology is an issue of importance. Putative C-terminal fragments from sortilin, a member of the vacuolar protein sorting 10 protein (Vps10p) family, have recently been shown to deposit in the neuritic ß-amyloid (Aß) plaques in the human brain. METHODS: We set out to explore if extracellular sortilin neuropathology exists in AD-related transgenic mice and nonhuman primates. Brains from different transgenic strains and ages developed overt cerebral Aß deposition, including the ß-amyloid precursor protein and presenilin 1 double-transgenic (APP/PS1) mice at ~ 14 months of age, the five familial Alzheimer's disease mutations transgenic (5×FAD) mice at ~ 8 months, the triple-transgenic Alzheimer's disease (3×Tg-AD) mice at ~ 22 months, and aged monkeys (Macaca mulatta and Macaca fascicularis) were examined. Brain samples from young transgenic mice, middle-aged/aged monkeys, and AD humans were used as negative and positive pathological controls. RESULTS: The C-terminal sortilin antibody, which labeled senile plaques in the AD human cerebral sections, did not display extracellular immunolabeling in the transgenic mouse or aged monkey brain sections with Aß deposition. In Western blot analysis, sortilin fragments ~ 15 kDa were not detectable in transgenic mouse cortical lysates, but they occurred in control AD lysates. CONCLUSIONS: In reference to their human brain counterparts, neuritic plaques seen in transgenic AD model mouse brains represent an incomplete form of this AD pathological hallmark. The species difference in neuritic plaque constituents also indicates more complex secondary proteopathies in the human brain relative to rodents and nonhuman primates during aging and in AD.

[Genetic diversity of germplasm resources of Lonicera japonica by AFLP analysis].

OBJECTIVE: This study aimed to analyze the genetic diversity and genetic relationship of germplasm resources of Lonicera japonica in main producing areas of China and provide reference for developing new varieties of L. japonica. METHOD: Using 6 primer combinations, 13 germplasm of L. japonica were analyzed by AFLP marker. The genetic distance was worked out by using DPS V3.01 software, and the cluster was conducted based on UPGMA. RESULT: A total of 435 bands were obtained including 191 polymorphic ones. The average polymorphic frequency was 43.9%. Cluster analysis showed that the relationship of cultivated variety from the same genuine area was near, and the classification result based on AFLP marker of germplasm of L. japonica from Shandong province was basically consistent with those on their morphological character. CONCLUSION: AFLP marker can indicate the abundant genetic diversity of L. japonica and provide theoretical evidence for reasonable utilization and breeding new cultivar of L. japonica in molecular level.

[Comparative study on morphological character of leaves of various farm cultivars of Lonicera japonica].

OBJECTIVE: To provide theoretical evidence for dividing and breeding cultivars of Lonicera japonica, the botanical character of laminas leaves observed and compared. METHOD: Using the morphological method, the main character of leaves of 11 farm cultivars of L. japonica were systematically observed and the data were comparatively studied with statistical means. It included the length, width and the ratio of the length to the width of blade, density, length and thickness of the nonglandular hair in the epidermis. RESULT: The results showed that there were obvious differences in length, width and the ratio of the length to the width of blade, density and length of the nonglandular hair in the epidermis. CONCLUSION: On the basis of the differences of morphological character of leaves, the various farm cultivars could be identified.

[Observation and comparision on morphological characteristics of pollen of Lonicera japonica in different cultivars].

OBJECTIVE: To compare morphological characteristics of pollen of Lonicera japonica in different cultivars cultivated in Shandong and provide a basis for distinguishing different cultivars and selecting fine breeding. METHOD: The scanning electron microscope (SEM) was applied to observe and compare the outside characteristics of pollen,and the data were analyzed by SAS 9.1 software. RESULT: Some difference was found among pollen size, aperture length and the density of spine. The biggest pollen size and aperture length is Dajizhao with 61.97 microm polar axis and 61.79 microm quarter major axis and 18.03 microm aperture length,and the smallest is Honggengzi with 57.46 microm polar axis and 57.29 microm quarter major axis and 18.03 microm aperture length. CONCLUSION: The morphological characteristics of pollen can provide a basis for distinguishing different cultivars of L. japonica.

[GC-MS analysis of volatile oil of olibanum].

OBJECTIVE: To analyze the volatile oil of olibanum and apply scientific evidences for its applications. METHOD: The volatile oil was analyzed by GC-MS. RESULT: One hundred components were identified,accounting for 91.26% of the total volatile oil, and the main components were octyl acetate, beta-elemene. It contains some transdermal absorption enhancers. CONCLUSION: The components of olibanum volatile oil were complicated; the connatural transdermal absorption enhancers make it possible to use in external preparation.

[Comparison of tanshinone IIA in Radix Salviae Miltorrhizae from Shandong].

OBJECTIVE: To compare the contents of tanshinone IIA in Radix Salviae Miltorrhizae from different species, production places and collecting time, for study its quantity from Shandong. METHODS: HPLC method was used. RESULTS: The contents of tanshinone IIA in Radix Salviae Miltorrhizae of Shandong ranged from 0.330%-0.726%. CONCLUSION: Setting the contents of tanshinone IIA as index, the quantity of Salvia miltiorrhiza Bge. f. alba C. Y. Wu et H. W. L. is better than Salvia miltiorrhiza Bge.. The genuine production place-Mengyin is the best planting area; the plantable place is mountaing district hillyland. Planting in plain region is inadvisable; the best collecting time is middle September.

[Studies on morphology and histology of Pinellia pedatisecta].

OBJECTIVE: To provide evidence for identification of the morphology and histology of Pinellia pedatisecta Schott. METHODS: Using microscope and scanning electron microscope, characteristics of the Pinellia pedatisecta Schott. were studied comparatively. RESULTS: The surface of leaves, fruits, testa and pollen grain of the cultivated Pinellia pedatisecta Schott. showed different characteristics. CONCLUSION: The results of the study can be used for the identification of the Pinellia pedatisecta Schott. in Shandong.

[Studies on micromorphological characters of fruit coats in cultivated Fructus Trichosanthis in Shandong Province].

OBJECTIVE: To explore the affinity relation among land race of Trichosanthes kirilowii and to provide evidence tor the classification and authentication. METHOD: Using scanning electron microscope, characteristics of the pericarp surface were studied comparatively. RESULT: The pericarp surface of the cultivated Fructus Trichosanthis in Shandong showed the characteristics of generality and diversity. CONCLUSION: The results of the study could be used for the identification of the cultivated Fructus Trichosanthis in Shandong.