Recent progress and challenges in the treatment of spinal cord injury

Spinal cord injury (SCI) disrupts the structural and functional connectivity between the higher center and the spinal cord, resulting in severe motor, sensory, and autonomic dysfunction with a variety of complications. The pathophysiology of SCI is complicated and multifaceted, and thus individual treatments acting on a specific aspect or process are inadequate to elicit neuronal regeneration and functional recovery after SCI. Combinatory strategies targeting multiple aspects of SCI pathology have achieved greater beneficial effects than individual therapy alone. Although many problems and challenges remain, the encouraging outcomes that have been achieved in preclinical models offer a promising foothold for the development of novel clinical strategies to treat SCI. In this review, we characterize the mechanisms underlying axon regeneration of adult neurons and summarize recent advances in facilitating functional recovery following SCI at both the acute and chronic stages. In addition, we analyze the current status, remaining problems, and realistic challenges towards clinical translation. Finally, we consider the future of SCI treatment and provide insights into how to narrow the translational gap that currently exists between preclinical studies and clinical practice. Going forward, clinical trials should emphasize multidisciplinary conversation and cooperation to identify optimal combinatorial approaches to maximize therapeutic benefit in humans with SCI.

Acquisition of Streptomycin Resistance by Oxidative Stress Induced by Hydrogen Peroxide in Radiation-Resistant Bacterium Deinococcus geothermalis.

Streptomycin is used primarily to treat bacterial infections, including brucellosis, plague, and tuberculosis. Streptomycin resistance easily develops in numerous bacteria through the inhibition of antibiotic transfer, the production of aminoglycoside-modifying enzymes, or mutations in ribosomal components with clinical doses of streptomycin treatment. (1) Background: A transposable insertion sequence is one of the mutation agents in bacterial genomes under oxidative stress. (2) Methods: In the radiation-resistant bacterium Deinococcus geothermalis subjected to chronic oxidative stress induced by 20 mM hydrogen peroxide, active transposition of an insertion sequence element and several point mutations in three streptomycin resistance (SmR)-related genes (rsmG, rpsL, and mthA) were identified. (3) Results: ISDge6 of the IS5 family integrated into the rsmG gene (dgeo_2335), called SrsmG, encodes a ribosomal guanosine methyltransferase resulting in streptomycin resistance. In the case of dgeo_2840-disrupted mutant strains (S1 and S2), growth inhibition under antibiotic-free conditions was recovered with increased growth yields in the presence of 50 µg/mL streptomycin due to a streptomycin-dependent (SmD) mutation. These mutants have a predicted proline-to-leucine substitution at the 91st residue of ribosomal protein S12 in the decoding center. (4) Conclusions: Our findings show that the active transposition of a unique IS element under oxidative stress conditions conferred antibiotic resistance through the disruption of rsmG. Furthermore, chronic oxidative stress induced by hydrogen peroxide also induced streptomycin resistance caused by point and frameshift mutations of streptomycin-interacting residues such as K43, K88, and P91 in RpsL and four genes for streptomycin resistance.

[Spatiotemporal evolution of habitat quality in three basins of Hainan Island based on InVEST model]. / 基于InVEST模型的海南岛三大流域生境质量时空演变.

Based on land use data of five periods during 1980 to 2020, using the InVEST model and the methods of land use transfer, habitat quality change rate and spatial statistical analysis, we explored the changes of habitat quality and its spatial distribution characteristics in the three major basins of Hainan Island (Nandu River, Changhua River and Wanquan River). The results showed that woodlands were the main land use type in the three basins of Hainan Island, accounting for more than 70% of the total area. From 1980 to 2020, the area of construction land increased the most, reaching up to 169.09 km2, mainly from cultivated land and woodland. The spatial distribution pattern of habitat quality in the study area was higher in the upstream and head water areas and lower in the mid and downstream regions. Overall, habitat quality index increased slightly for a short period and then decreased significantly during the study period. Among the three basins, habitat quality of Wanquan River Basin was the highest, followed by Changhua River Basin, and Nandu River Basin was the lowest. The habitat quality of Nandu River Basin fluctuated greatly and was strongly affected by human disturbance. From 1980 to 2020, the change rate of habitat quality in the three basins generally decreased by 0.5%, which was significantly degraded from 2010 to 2020. From 1980 to 2020, the spatial distribution of habitat quality in the study area displayed strong autocorrelation and significant aggregation. The hot spot area of habitat quality was mainly concentrated near the head water and upstream areas of the three basins, while the cold spot area was mainly distributed in the estuary area of the three basins, along with the mid and downstream areas of the Nandu River. These results would provide scientific reference for biodiversity conservation and ecological restoration efforts in the three basins of Hainan Island.

Identification of novel biomarkers and therapeutic target candidates for stasis-heat symptom pattern of acute intracerebral hemorrhage by quantitative plasma proteomics.

OBJECTIVE: To explore the novel biomarkers and therapeutic target candidates related to the stasis-heat syndrome of acute intracerebral hemorrhage (AICH). METHODS: Applying an isobaric tagging for relative and absolute quantitation-(iTRAQ-) based quantitative proteomic approach, plasma samples from AICH patients with stasis-heat, and AICH patients with non-stasis-heat and healthy control subjects were collected and analyzed to distinguish differentially expressed proteins (DEPs) correlated to AICH with stasis-heat in this block design. The standard Western blot was applied to verify DEPs. Additionally, DEPs were analyzed via bioinformatic platforms and further approved via Ingenuity Pathway Analysis (IPA). RESULTS: A total of 26 DEPs were found among AICH with the stasis-heat, AICH with non-stasis-heat, and healthy control group. The seven DEPs compared with the non-stasis-heat group are closely related to the pathogenesis of stasis heat. These proteins showed three different protein expression patterns. The alpha-1-b glycoprotein (A1BG) and copper-protein (CP) were up-regulated in the stasis-heat group, but down-regulated in the non-stasis-heat group. Compared with the non-stasis-heat group, the expression abundance of actinin, alpha 1 (ACTN1), carbonic anhydrase I (CA1), peroxiredoxin 2 (PRDX2), and vinculin (VCL) is higher in the stasis-heat group, while the CD44 is the opposite. These differences reflect that stasis-heat syndrome has more severe inflammatory immune response, coagulation disorders and damage. Bioinformatics analysis revealed that a wide variety of cellular and metabolic processes and some signaling pathways were involved in the pathophysiology of AICH with stasis-heat. AICH with stasis-heat syndrome showed more severe inflammatory reactions, tissue damage, and coagulation disorders than non-stasis heat syndrome. CONCLUSIONS: There are differences in the protein expression patterns between the stasis-heat syndrome and non-stasis-heat syndrome. These differences reflect that stasis-heat syndrome has more severe damage. CD44, CP, ACTN1, CA1, VCL, PRDX2, and A1BG could be the potential biomarkers and therapeutic target candidates of the stasis-heat subtype. This study provides a reasonable explaination for Liangxue Tongyu decoction through anti-inflammatory and brain protection treatment.

The protective effect of fluoxetine hydrochloride on brain tissues of rats with high altitude cerebral edema / 中国药理学通报

Aim To study the protective effect of fluoxetine hydrochloride on brain tissues of rats with simulated high altitude cerebral edema(HACE)and its mechanism.Methods The optimal dosage and time of fluoxetine hydrochloride were determined by the hypoxia tolerance test of mice under normal pressure.The rat model of brain edema at high altitude was established by large-scale low-pressure oxygen chamber.HE staining was used to observe the pathological changes of brain tissues in rats.Microplate reader was used to detect the corresponding indexes of oxidative stress such as malondialdehyde(MDA)level and superoxide dismutase(SOD)activity.The expressions of hypoxia-related proteins HIF-1α,VEGF,MMP-9,AQP4 and SERT were detected by Western blot.Results Compared with the hypoxia model group,after the intervention of fluoxetine hydrochloride,the survival time of mice was prolonged,and the middle dose of fluoxetine(14 mg·kg-1)had the best effect,with an extension rate of 17.78%.The pathological damage of brain was improved,the water content of brain decreased,and the permeability of blood-brain barrier decreased.MDA content in rat brain decreased and SOD activity increased.Western blot results showed that HIF-1α,VEGF,MMP-9,AQP4,SERT protein were significantly down-regulated.Conclusions Fluoxetine has protective effect on rats with brain edema at high altitude,and its mechanism may be related to improving oxidative stress,activating HIF-1α/VEGF/MMP-9 signaling pathway and affecting the expression of SERT protein.SERT may be a potential target for treating brain edema at high altitude.

Analysis and suggestions on harvest period standards of plant medicinal materials in 2020 edition of Chinese Pharmacopoeia / 中国中药杂志

The improvement of the harvest period standards is critical in the quality control of Chinese medicinal materials. The present study statistically analyzed the harvest period standards of plant medicinal materials in the 2020 edition of Chinese Pharmacopoeia(Vol.Ⅰ) and put forward the existing problems and suggestions based on herbal records and modern research to provide references for the improvement of the standards. According to the statistical analysis, in 499 types of plant medicinal materials, harvest period standards are recorded under 486 types, accounting for 97.4%, and are lacking in the remaining. Only one medicinal material(Stellariae Radix) is recorded with the standard of the harvest year. The standards of the harvest season and phenological period are recorded under 233 types, accounting for 46.7%. For 237 types, only harvest season is specified, accounting for 47.5%, and for 15 types, only harvest phenological period is specified, accounting for 3.0%. Among 222 types mainly derived from cultivation and 51 types from wild resources and cultivation, only 11 types are recorded with harvest period of cultivated products. Only Stellariae Radix is recorded with the harvest period standards for the wild and cultivated products separately. The harvest period standards of plant medicinal materials with different medicinal parts have certain rules to follow. The main problems about the harvest period standards are discovered. Specifically, no harvest period standards are recorded under 13 types of plant medicinal materials. Almost all perennial cultivated medicinal materials are not recorded with harvest year standard. No phenological period standard is found under 250 types of plant medicinal materials. There is no clear distinction between the harvest period standards of cultivated and wild products. The evidence for harvest period standards of 26 types of plant medicinal materials that can be harvested all year round is insufficient. As a result, it is proposed to strengthen basic research in response to the above-mentioned problems and improve the harvest period standards as soon as possible to ensure the quality of Chinese medicinal materials.

Gross ecosystem product accounting of national park: Taking Hainan Tropical Rainforest National Park as an example.

Hainan Tropical Rainforest National Park has the most representative and largest contiguous tropical rainforest in China, which has advantages in exploring the realization mechanism of ecological product value in national parks. Based on the basic framework of "The Technical Guideline on Gross Ecosystem Product (GEP)", we constructed a GEP accounting system in line with the characteristics of tropical rain forest national park, and calculated the GEP of Hainan Tropical Rainforest National Park in 2019. The results showed that the GEP of Hainan Tropical Rainforest National Park in 2019 was 204.513 billion yuan, and the GEP per unit area was 0.046 billion yuan·km-2. Among all the service types, the value of material services was 4.850 billion yuan, accounting for 2.4% of the total GEP in the national park. The ecosystem regulation service value was 168.891 billion yuan, accounting for 82.6%. The value of cultural services was 30.772 billion yuan, accounting for 15.0%. Among different ecosystem types, the unit area value of the tropical rain forest ecosystem represented by mountain rain forest, lowland rain forest, deciduous monsoon forest, and tropical cloud forest was much higher than that of plantation or other ecosystems, indicating the dominant role of tropical rain forest ecosystem in providing ecosystem services. In addition, based on the GEP accounting results of the national park, we put forward relevant suggestions for further exploring the realization path and realization mechanism of ecological product value.

Sulfite as the substrate of C-sulfonate metabolism of α, β-unsaturated carbonyl containing andrographolide: analysis of sulfite in rats' intestinal tract and the reaction kinetics of andrographolide with sulfite / 中国天然药物

One-sixth of the currently known natural products contain α, β-unsaturated carbonyl groups. Our previous studies reported a rare C-sulfonate metabolic pathway. Sulfonate groups were linked to the β-carbon of α, β-unsaturated carbonyl-based natural compounds through this pathway. However, the mechanism of this type of metabolism is still not fully understood, especially whether it is formed through enzyme-mediated biotransformation or direct sulfite addition. In this work, the enzyme-mediated and non-enzymatic pathways were studied. First, the sulfite content in rat intestine was determined by LC-MS/MS. The results showed that the amount of sulfite in rat intestinal contents was from 41.5 to 383 μg·g

Influence of dietary factors on radius speed of sound in female college students / 中国学校卫生

Objective@#To understand the level of radius speed of sound (SOS) in female college students and explore the effect of dietary on their bone health,so as to provide a reference for effective prevention fracture and osteoporosis among college students.@*Methods@#A total of 363 female college students were selected by multistage stratified random sampling method. Dietary intake was assessed with a self-administered food frequency questionnaire, principal component analysis was used to derive dietary patterns. Speed of sound was measured by ultrasonic bone densitometer.@*Results@#In all, the mean SOS of female college students was(4 138.0±114.3)m/s, 26.7% of female college students had low bone mass. Three dietary patterns were identified by factor analysis, explaining 55.4% of the diet variation: "snack" "plant" "high protein and high fat" patterns, respectively. Multiple linear regression analysis showed that with the increase of age, the level of SOS in radius increased significantly; and snacks consumption was negatively associated with the level of SOS(β=13.23, -0.47, P<0.05).@*Conclusion@#University stage is still the critical period of bone mass accumulation for female, snacks consumption may have a negative impact on bone health, so it is necessary to carry out nutrition education to help improve health awareness, promote health eating, and lead health lives to prevent osteoporosis.

The timing and effectiveness of implementing mild interventions of COVID-19 in large industrial cities

The outbreak of novel coronavirus disease (COVID-19) has spread around the world since it was detected in December 2019. The Chinese government executed a series of interventions to curb the pandemic. The "battle" against COVID-19 in Shenzhen, China is valuable because populated industrial cities are the epic centres of COVID-19 in many regions. We made use of synthetic control methods to create a reference population matching specific characteristics of Shenzhen. With both the synthetic and observed data, we introduced an epidemic compartmental model to compare the spread of COVID-19 between Shenzhen and its counterpart regions in the United States that didnt implement interventions for policy evaluation. Once the effects of policy interventions adopted in Shenzhen were estimated, the delay effects of those interventions were referred to provide the further control degree of interventions. Thus, the hypothetical epidemic situations in Shenzhen were inferred by using time-varying reproduction numbers in the proposed SIHR (Susceptible, Infectious, Hospitalized, Removed) model and considering if the interventions were delayed by 0 day to 5 days. The expected cumulative confirmed cases would be 1546, which is 5.75 times of the observed cumulative confirmed cases of 269 in Shenzhen on February 3, 2020, based on the data from the counterpart counties (mainly from Broward, New York, Santa Clara, Pinellas, and Westchester) in the United States. If the interventions were delayed by 5 days from the day when the interventions started, the expected cumulative confirmed cases of COVID-19 in Shenzhen on February 3, 2020 would be 676 with 95% credible interval (303,1959). Early implementation of mild interventions can subdue the epidemic of COVID-19. The later the interventions were implemented, the more severe the epidemic was in the hard-hit areas. Mild interventions are less damaging to the society but can be effective when implemented early. AMS 2000 O_SCPLOWSUBJECTC_SCPLOWO_SCPCAP C_SCPCAPO_SCPLOWCLASSIFICATIONSC_SCPLOW: Primary 00K00, 00K01; secondary 00K02.

COVID-Net: A deep learning based and interpretable predication model for the county-wise trajectories of COVID-19 in the United States

The cases of COVID-19 have been reported in the United States since January 2020. We propose a COVINet by combining the architecture of both Long Short-Term Memory and Gated Recurrent Unit. First, we use the 10-fold cross-validation to train and assess different prediction models for which all counties serve alternatively as the training and test counties. Then, we focus on the prediction for the 10 severest counties. We employ the Mean Relative Errors (MREs) to measure the performance of the COVINet in predicting confirmed cases and deaths. Two COVINet models with 26 and 19 input variables, respectively, are trained. We estimate their respective MREs in the last 30 days before January 23, 2021, by the 10-fold CV, which are 0.0898 and 0.1068 for the number of confirmed cases, and 0.0694 and 0.0724 for the number of deaths. The MREs are also small for all predictions of the events in the last 7 or 30 days before January 23, 2021. The COVINet uses features including workforce driving alone to work, traffic volume, income inequality, and longitude and latitude of infected counties to predict the trajectories of COVID-19 in counties of the United States. The increasing awareness of how predictors affect the pandemic helps policymakers develop plans to mitigate the spread of COVID-19.

Risk factors associated with mortality of COVID-19 in 2692 counties of the United States

BackgroundThe number of cumulative confirmed cases of COVID-19 in the United States has risen sharply since March 2020. A county health ranking and roadmaps program has been established to identify factors associated with disparity in mobility and mortality of COVID-19 in all counties in the United States. MethodsTo find out the risk factors associated with county-level mortality of COVID-19 with various levels of prevalence, a negative binomial design was applied to the county-level mortality counts of COVID-19 as of August 27, 2020 in the United States. In this design, the infected counties were categorized into three levels of infections using clustering analysis based on time-varying cumulative confirmed cases from March 1 to August 27, 2020. COVID-19 patients were not analyzed individually but were aggregated at the county-level, where the county-level deaths of COVID-19 confirmed by the local health agencies. Results3125 infected counties were assigned into three classes corresponding to low, median, and high prevalence levels of infection. Several risk factors were significantly associated with the mortality counts of COVID-19, where higher level of air pollution (0.153, P<0.001) increased the mortality in the low prevalence counties and elder individuals were more vulnerable in both the median and high prevalence counties. The segregation between non-Whites and Whites and higher Hispanic population had higher likelihood of risk of the deaths in all infected counties. ConclusionsThe mortality of COVID-19 depended on sex, race/ethnicity, and outdoor environment. The increasing awareness of the impact of these significant factors may lead to the reduction in the mortality of COVID-19.

Evaluate the timing of resumption of business for the states of New York, New Jersey and California via a pre-symptomatic and asymptomatic transmission model of COVID-19

The United States has the highest numbers of confirmed cases of COVID-19 in the world. The early hot spot states were New York, New Jersey, and Connecticut. The workforce in these states was required to work from home except for essential services. It was necessary to evaluate an appropriate date for resumption of business since the premature reopening of the economy would lead to a broader spread of COVID-19, while the opposite situation would cause greater loss of economy. To reflect the real-time risk of the spread of COVID-19, it was crucial to evaluate the population of infected individuals before or never being confirmed due to the pre-symptomatic and asymptomatic transmissions of COVID-19. To this end, we proposed an epidemic model and applied it to evaluate the real-time risk of epidemic for the states of New York, New Jersey, and Connecticut. We used California as the benchmark state because California began a phased reopening on May 8, 2020. The dates on which the estimated numbers of unidentified infectious individuals per 100,000 for states of New York, New Jersey, and Connecticut were close to those in California on May 8, 2020, were June 1, 22, and 22, 2020, respectively. By the practice in California, New York, New Jersey, and Connecticut might consider reopening their business. Meanwhile, according to our simulation models, to prevent resurgence of infections after reopening the economy, it would be crucial to maintain sufficient measures to limit the social distance after the resumption of businesses. This precaution turned out to be critical as the situation in California quickly deteriorated after our analysis was completed and its interventions after the reopening of business were not as effective as those in New York, New Jersey, and Connecticut.

The Effects of Stringent Interventions for Coronavirus Pandemic

The pandemic of COVID-19 has caused severe public health consequences around the world. Many interventions of COVID-19 have been implemented. It is of great public health and societal importance to evaluate the effects of interventions in the pandemic of COVID-19. In this paper, with help of synthetic control method, regression discontinuity and a Susceptible-Infected and infectious without isolation-Hospitalized in isolation-Removed (SIHR) model, we evaluate the horizontal and longitudinal effects of stringent interventions implemented in Wenzhou, a representative urban city of China, where stringent interventions were enforced to curb its own epidemic situation with rapidly increasing newly confirmed cases. We found that there were statistically significant treatment effects of those stringent interventions which reduced the cumulative confirmed cases of COVID-19. Those reduction effects would increase over time. Also, if the stringent interventions were delayed by 2 days or mild interventions were implemented instead, the expected number of cumulative confirmed cases would have been nearly 2 times or 5 times of the actual number. The effects of stringent interventions are significant in mitigating the epidemic situation of COVID-19. The slower the interventions were implemented, the more severe the epidemic would have been, and the stronger the interventions would have been required.

Data-driven analysis on the simulations of the spread of COVID-19 under different interventions of China

Since February 2020, COVID-19 has spread rapidly to more than 200 countries in the world. During the pandemic, local governments in China have implemented different interventions to efficiently control the spread of the epidemic. Characterizing transmission of COVID-19 under some typical interventions is essential to help countries develop appropriate interventions. Based on the pre-symptomatic transmission patterns of COVID-19, we established a novel compartmental model: Baysian SIHR model with latent Markov structure, which treated the numbers of infected and infectious individuals without isolation to be the latent variables and allowed the effective reproduction number to change over time, thus the effects of policies could be reasonably estimated. By using the epidemic data of Wuhan, Wenzhou and Shenzhen, we migrated the corresponding estimated policy modes to South Korea, Italy, and the United States and simulated the potential outcomes for these countries when they adopted similar policy strategies of three cities in China. We found that the mild interventions implemented in Shenzhen were effective to control the epidemic in the early stage, while more stringent policies which were issued in Wuhan and Wenzhou were necessary if the epidemic was more severe and needed to be controlled in a short time.

Effect of Signal Transduction Pathway Gene Mutations on the One- course Induced Remission Rate and Analysis of Clinical Characteristics in Patients with CBF-AML / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of other gene mutations outside the fusion gene on the first complete remission (CR) induced by one course of induction chemotherapy in patients with core binding factor-associated acute myeloid leukemia (CBF-AML).@*METHODS@#DNA was extracted from bone marrow or peripheral blood samples of newly diagnosed CBF-AML patients admitted to the Hematology Department of the Second Hospital of Shanxi Medical University from January 2015 to January 2019. Next-generation sequencing was used for detection of 34 kinds of hematologic malignancy-related gene mutations in patients with CBF-AML, the effect of related gene mutations on the first complete remission (CR) rate in one course of induction chemotherapy was analyzed by combineation with clinical characteristics.@*RESULTS@#34 kinds of genes in bone marrow or peripheral blood of 43 patients were detected by high throughput sequencing and the gene mutations were detected in 16 out of 34 genes. The mutation rate of KIT gene was the highest (48.8%), followed by NRAS (16.3%), ASXL1 (16.3%), TET2 (11.6%), CSF3R (9.3%), FLT3 (9.3%), KRAS (7.0%). The detection rates of mutations in different functional genes were as follows: genes related with signal transduction pathway (KIT, FLT3, CSF3R, KRAS, NRAS, JAK2, CALR, SH2B3, CBL) had the highest mutation frequency (72.1% (31/43); epigenetic modification gene mutation frequency was 30.2% (13/43), including ASXL1, TET2, BCOR); transcriptional regulation gene mutation frequency was 7.0% (3/43), including ETV6, RUNX1, GATA2). Splicing factor related gene mutation frequency was 2.3% (1/43), including ZRSR2). The CR rate was 74.4% after one course of induction chemotherapy. At first diagnosis, patients with low expression of WT1 (the median value of WT1 was 788.9) were more likely to get CR (P=0.032) and the RFS of patients who got CR after one course of induction chemotherapy was significantly longer than that of patients without CR [7.6 (2.2-44.1) versus 5.8 (1-19.4), (P=0.048)]. The rate of CR in the signal transduction pathway gene mutation group was significantly lower than that in non-mutation group (64.5% vs 100%) (P=0.045), while the level of serum hydroxybutyrate dehydrogenase (HBDH) was significantly higher than that in non-mutation group [(418 (154-2702) vs 246 (110-1068)] (P=0.032). There was no difference in CD56 expression between the two groups (P=0.053), which was limited to the difference between (≥20%) expression and non-expression. (P=0.048).@*CONCLUSION@#CBF-AML patients with signal transduction pathway gene mutation are often accompanied by high HBDH level and CD56 expression, moreover, the remission rate induced by one course of treatment is low.

LCMS-IT-TOF Analysis of Retention Behavior of Sanguisorbae Radix Components on D101 Macroporous Resin and Polyamide Resin / 中国实验方剂学杂志

Objective:To study on the material basis of Sanguisorbae Radix by column chromatography and liquid chromatography-ion trap-time-of-flight mass spectrometry (LCMS-IT-TOF), and analyze the distribution of different components in Sanguisorbae Radix water extract on D101 macroporous resin and polyamide resin. Method:Sanguisorbae Radix water extract was separated by D101 macroporous resin and polyamide resin, and LCMS-IT-TOF was used for detection, chromatography separation was achieved on an ACQUITY UPLC HSS T3 column (2.1 mm×100 mm, 1.8 μm) with the mobile phase consisted of water (A) and acetonitrile (B) for gradient elution (0-10 min, 5%-20%B; 10-18 min, 20%-35%B; 18-23 min, 35%-50%B; 23-28 min, 50%-90%B; 28-30 min, 90%B; 30-33 min, 90%-5%B; 33-35 min, 5%B), the flow rate was 0.3 mL·min-1, the column temperature was 30 ℃. Data acquisition was carried out in electrospray ionization (ESI) under the positive and negative ion modes, the scanning range was m/z 100-1 200. According to mass spectrometry data such as accurate molecular mass and fragment information, combined with literature, different chemical components in loading effluents and ethanol eluents of Sanguisorbae Radix water extract were identified. A heat map of the distribution of components in each fraction was drawn by extracting mass spectrum peak intensity data of each sample. The elution rules of various components were compared visually. Result:The enrichment and separation of D101 macroporous resin and polyamide resin were obvious. Tannins in Sanguisorbae Radix water extract was mainly concentrated in loading effluent of macroporous resin and its water eluent, triterpenoids were mainly distributed in the 90% ethanol eluent of macroporous resin. In the above effluents and eluents, a total of 63 compounds (including isomers) were identified. Among them, 6 compounds, ellagic acid-4-pyranoarabinoside or its isomer, 6-O-galloylnorbergerin, 3-O-galloylnorbergerin, (6-acetyloxy-5,7-dihydroxy-8-methoxy-4-oxochromen-2-yl) acetate, ethyl 2-methyl-5,6-bis (sulfooxy) benzofuran-3- carboxylate were first discovered in Sanguisorbae Radix. Conclusion:The method can quickly and accurately identify the distribution of components in aqueous extract of Sanguisorbae Radix after column chromatography, providing experimental basis for exploring the pharmacodynamic components and mechanism of Sanguisorbae Radix.

Research and application of tissue optical clearing technique / 中国组织工程研究

BACKGROUND: Whole-body cell analysis of organisms is one of the major challenges in biomedicine. Tissue optical clearing technique combined with optical imaging and image processing technique can make the whole organ or the body transparent rapidly for structural and cellular analyses, providing a very promising solution for the application of advanced optical technique in life science. OBJECTIVE: To analyze the principle and process of tissue optical clearing technique, to summarize the research progress of tissue optical clearing technique, to present imaging technology for tissue clearing, and to discuss the application of tissue optical clearing technique in biomedical researches. METHODS: The first author searched relevant literatures in PubMed database with the keywords of “tissue optical clearing technique, tissue optical clearing, whole-body imaging and 3D imaging”. A total of 168 articles were initially retrieved. After sorting and screening systematically, 72 literatures were included for analysis, summary and discussion. RESULTS AND CONCLUSION: Current tissue optical clearing protocols are divided into two groups: solvent-based clearing methods and hydrophilic reagent-based clearing methods. Tissue clearing is usually conducted by the following steps: (a) tissue fixation, (b) permeabilization, (c) decolorization, and (d) refractive index matching. Tissue optical clearing technique can rapidly transform tissue into an optically transparent form, improving imaging depth and contrast. Combined with microscope imaging techniques such as confocal, two-photon and light sheet microscopes, tissue clearing can achieve 3D imaging of the whole organ or body at cellular resolution, accelerating the process of whole-body cell analysis of organisms. Tissue optical clearing technique will continue to evolve in the future, promote the development of new clearing reagents and clearing optimized microscopes, further enhance the acquisition of structural and molecular information from intact systems and contribute to the comprehensive understanding of whole biological systems.

Clinical effect of nano-fat mixed granule fat transplantation in the treatment of cicatricial facial depression and atrophy and the related experimental mechanism / 中华烧伤杂志

Objective@#To study the clinical effect of nano-fat mixed granule fat transplantation in the treatment of cicatricial facial depression and atrophy, and to explore the related experimental mechanism.@*Methods@#(1) From January 2012 to April 2018, 105 patients conforming to the inclusion criteria, with cicatricial facial depression and atrophy deformity who needed facial fat transplantation, were admitted to our unit. Their medical records were analyzed retrospectively. According to the patients′ wishes, 54 patients (12 males and 42 females) aged 10-59 years received traditional transplantation of pure autologous granule fat collected from abdomen/thigh and were included in simple transplantation group, while the other 51 patients (14 males and 37 females) aged 7-63 years received transplantation of autologous nano-fat mixed granule fat collected from abdomen/thigh and were included in mixed transplantation group. The treatment satisfaction of patients 3 and 6 months after operation was assessed by the facial fullness, symmetry, scar, and complications using self-made scales and photo data before and after operation. Six months after operation, the patients were assessed whether they needed to undergo a second operation, and the rate of second operation was calculated. During the second operation, the fat of patients transplanted in the first operation was collected, and the morphology of adipocytes and microangiogenesis was observed under a scanning electron microscope. (2) Adipose-derived stem cells (ADSCs) were isolated and cultured from abdominal fat of a 4-week-old male Sprague-Dawley (SD) rat. The 5th passage of cells were selected to observe cell morphology after cultured for 14 days, observe expression of vimentin and cytokeratin-18 by immunofluorescence method, identify osteogenic and adipogenic differentiation, and detect rates of CD29 and CD44 positive cells by flow cytometer (n=3). Eighteen 4-week-old male SD rats were divided into ADSCs transplantation group, simple scar group, and blank control group according to the random number table, with 6 rats in each group. Rats in ADSCs transplantation group and simple scar group were subcutaneously injected with 1 mL bleomycin which was dissolved in phosphate buffered saline (PBS) with a mass concentration of 1 mg/mL at the back to establish scar models. After 3 hours, rats in ADSCs transplantation group were injected with 1×106 ADSCs suspended in 0.1 mL PBS at the same injection site, while rats in simple scar group were injected with 0.1 mL PBS. Rats in blank control group were injected with the same doses of PBS in the same place at the same two time points mentioned above. After continuous injection for 28 days in each group, the full-thickness skin tissue of the injected area of all rats was collected to observe the collagen fibers by Masson staining and expressions of α-smooth muscle actin (α-SMA) and transforming growth factor β1 (TGF-β1) by immunohistochemistry, and the positive cells were counted. Data were processed with Mann-Whitney U test, χ2 test, one-way analysis of variance, and least significant difference test.@*Results@#(1) Compared with the preoperative condition, the facial fullness and symmetry of patients in simple transplantation group were better in 3 months after operation, with scar color closer to the surrounding skin, and the filling volume of patients in this group decreased in 6 months after operation as compared with that in 3 months after operation. In mixed transplantation group, the facial fullness and symmetry of patients were better in 3 and 6 months after operation as compared with the preoperative condition, with scar color and texture closer to the surrounding skin, and the filling volume in 6 months after operation was not obviously reduced as compared with that in 3 months after operation. Fat liquefaction and subcutaneous nodule formation occurred respectively in 1 patient in simple transplantation group within 3 months after operation. The treatment satisfaction of patients in mixed transplantation group was significantly higher than that in simple transplantation group in 3 and 6 months after operation (Z=-2.566, -3.084, P<0.05 or P<0.01). Six months after operation, the second operation rate of patients in mixed transplantation group was 7.84% (4/51), which was significantly lower than 22.22% (12/54) in simple transplantation group (χ2=4.199, P<0.05). At the second operation, compared with those of simple transplantation group, the cells of fat transplanted in the first operation of patients in mixed transplantation group were more plump, without collapse or dryness, and the cells were closely arranged, with smaller gap; the tubular and the cord-like microvascular structure on the cell surface were more abundant, and the cell gap was full of network-like microvascular structure that grew into the adipose tissue. (2) The fifth passage of cells isolated and cultured from rat fat grew adherently to the wall, with long fusiform or spindle shape, showing shoal-of-fish-like growth. Vimentin and cytokeratin-18 were highly expressed in the cells. Cells showed osteogenic and adipogenic differentiation ability by induction. The positive expression rates of CD29 and CD44 were higher than 90.00%. The cells were identified as ADSCs. After 28 days of injection, the collagen fibers in the dermis of skin tissue at the injection area of rats in blank control group were finely arranged. In simple scar group, a large amount of collagen was deposited in the dermis of skin tissue at the injection area of rats, the fiber bundles were thick and loosely unevenly arranged, and a large number of inflammatory infiltration and scattered muscle fibers were observed. In ADSCs transplantation group, the collagen fibers in the dermis of skin tissue at the injection area of rats were thicker than those of blank control group, with still neat arrangement, and a small amount of scattered muscle fiber and inflammatory infiltration was observed. After 28 days of injection, the expression of α-SMA in ADSCs transplantation group was mainly in microvessels in the dermis of skin tissue at the injected area of rats, and the number of α-SMA and TGF-β1 positive cells was (49±12) and (63±10) cells per 20-fold field of view, respectively, which was similar to (35±16) and (44±17) cells per 20-fold field of view of blank control group (P>0.05), all significantly less than (135±13) and (121±23) cells per 20-fold field of view of simple scar group (P<0.05).@*Conclusions@#Compared with those of autologous simple granule fat transplantation, autologous nano-fat mixed granule fat transplantation has better filling fullness in the treatment of patients with scar facial depression and atrophy. The filling effect lasts longer, and the improvement of scar texture is more obvious. As showed in the rat scar model experiment, the mechanism may be that ADSCs inhibit the expressions of α-SMA and TGF-β1, thus inhibiting the formation of scar.

Reaction of Chlorogenic Acid with Sulfite and Its Application in Identification of Sulfur-fumigated Herbs / 中国实验方剂学杂志

Objective: To explore the structure and reaction sites of the reaction products of chlorogenic acid and sodium sulfite,and the chemical changes of chlorogenic acid in Lonicerae Japonicae Flos after sulfur fumigating. Method: Chlorogenic acid was reacted with sodium sulfite under mild conditions. Liquid chromatography-mass spectrometry-ion trap-time-of-flight (LC-MS-IT-TOF) and 1H nuclear magnetic resonance spectroscopy (1H-NMR) detection techniques were used to detect the reaction products,and the sulfur-fumigated and unsulfurized Lonicerae Japonicae Flos water extract was detected by LC-MS-IT-TOF. Result: After analyzing the mass spectrometry data of fragment ion,molecular cleavage and accurate molecular weight,according to the results of nuclear magnetic signals of chemical shift,peak intensity and peak splitting, the products of chlorogenic acid and sodium sulfite were preliminarily identified as chlorogenic acid α,β-unsaturated carbonyl addition product:3-((3-(3,4-dihydroxyphenyl)-2-sulfopropyl)oxy)-1,4,5-trihydroxycyclohexane-1-carboxylic acid or 3-((3-(3,4-dihy droxyphenyl)-3-sulfopropyl)oxy)-1,4,5-trihydroxycyclohexane-1-carboxylic acid,and the same characteristic fragments were detected as the addition product in the sulfur fumigated Lonicerae Japonicae Flos,but not found in the unsulfurized. Conclusion: It is the first time to demonstrate the structure and reaction sites of chlorogenic acid and sulfurous acid reaction products,and detect the chlorogenic acid sulfite addition product in sulfur-fumigated Lonicerae Japonicae Flos. Although it is still unclear how the sulfite addition compound produced by sulphur Lonicerae Japonicae Flos affects the efficacy and toxicological activity of Lonicerae Japonicae Flos,we shall still pay attention to the changes of active ingredients in sulphuric medicinal materials. Besides,this study can also provide reference for the studies of chemical composition changes after sulfuration of traditional Chinese medicine containing α,β-unsaturated carbonyl structure.