Single-Cell Transcriptome Landscape and Cell Fate Decoding in Human Brain Organoids after Transplantation.

Human stem cells and derivatives transplantation are widely used to treat nervous system diseases, while the fate determination of transplanted cells is not well elucidated. To explore cell fate changes of human brain organoids before and after transplantation, human brain organoids are transplanted into prefrontal cortex (PFC) and hippocampus (HIP), respectively. Single-cell sequencing is then performed. According to time-series sample comparison, transplanted cells mainly undergo neural development at 2 months post-transplantation (MPT) and then glial development at 4MPT, respectively. A different brain region sample comparison shows that organoids grafted to PFC have obtained cell fate close to those of host cells in PFC, other than HIP, which may be regulated by the abundant expression of dopamine (DA) and acetylcholine (Ach) in PFC. Meanwhile, morphological complexity of human astrocyte grafts is greater in PFC than in HIP. DA and Ach both activate the calcium activity and increase morphological complexity of astrocytes in vitro. This study demonstrates that human brain organoids receive host niche factor regulation after transplantation, resulting in the alignment of grafted cell fate with implanted brain regions, which may contribute to a better understanding of cell transplantation and regenerative medicine.

[Spatiotemporal Simulation and Prediction of Grassland Carbon Budget in Gansu Under Different Climate Scenarios].

Under the background of carbon peak and carbon neutrality, grassland carbon sinks are a key pathway to carbon neutrality. Based on the Coupled Model Intercomparison Project Phase 6 (CMIP6) climate scenario data, the Daycent model was used to simulate the carbon budget of Gansu grasslands from 2015 to 2100, and the trend analysis was used to study the spatial and temporal changes in grassland carbon budget in the next 78 years. The results revealed that, under the future climate scenario of SSP245, the net ecosystem productivity (NEP) indicated a non-significant fluctuating downward trend with a rate of -0.20 g·(m2·a)-1(in C, the same below), and the grassland carbon sink was in a declining state. Under the future climate scenario of SSP585, the grassland NEP indicated a significant fluctuating increase trend with a growth rate of 1.36 g·(m2·a)-1, and the grassland carbon sink gradually increased under this scenario; the spatial distribution of grassland carbon budget increased from northwest to southeast. The increase in temperature and precipitation under the SSP585 climate scenario was higher than that under the SSP245 climate scenario, and the grassland carbon budget strongly correlated positively with precipitation. However, a negative correlation was observed between grassland carbon budget and temperature. We identified the carbon sink intensity in Gansu grasslands under different climate conditions, which provides a reference for and contribution to effective carbon sequestration.

Latent class cluster analysis of knowledge on acute myocardial infarction in community residents: a cross-sectional study in Tianjin, China.

OBJECTIVE: Public knowledge of early onset symptoms and risk factors (RF) of acute myocardial infarction (AMI) is very important for prevention, recurrence and guide medical seeking behaviours. This study aimed to identify clusters of knowledge on symptoms and RFs of AMI, compare characteristics and the awareness of the need for prompt treatment. DESIGN: Multistage stratified sampling was used in this cross-sectional study. Latent GOLD Statistical Package was used to identify and classify the respondent subtypes of the knowledge on AMI symptoms or modifiable RFs. Multivariable logistic regression was performed to identify factors that predicted high knowledge membership. PARTICIPANTS: A structured questionnaire was used to interview 4200 community residents aged over 35 in China. 4122 valid questionnaires were recovered. RESULTS: For AMI symptoms and RFs, the knowledge levels were classified into two or three distinct clusters, respectively. 62.7% (Symptom High Knowledge Cluster) and 39.5% (RF High Knowledge Cluster) of the respondents were able to identify most of the symptoms and modifiable RFs. Respondents who were highly educated, had higher monthly household income, were insured, had regular physical examinations, had a disease history of AMI RFs, had AMI history in immediate family member or acquaintance or had received public education on AMI were observed to have higher probability of knowledge on symptoms and RFs. There was significant difference in awareness of the prompt treatment in case of AMI occurs among different clusters. 'Calling an ambulance' was the most popular option in response of seeing others presenting symptoms of AMI. CONCLUSIONS: A moderate or relatively low knowledge on AMI symptoms and modifiable RFs was observed in our study. Identification of Knowledge Clusters could be a way to detect specific targeted groups with low knowledge of AMI, which may facilitate health education, further reduce the prehospital delay in China and improve patient outcomes.

Lactobacillus levels and prognosis of patients with acute myocardial infarction.

OBJECTIVE: To explore the relationship between Lactobacillus and prognosis of acute myocardial infarction (AMI) patients treated by percutaneous coronary intervention (PCI) and its correlation with clinical parameters. METHODS: Consecutive patients with AMI in the coronary care unit of Tianjin Chest Hospital in China who received emergency PCI between July 2017 and December 2018 were enrolled. Subjects' fecal 16S rDNA gene sequencing data were analyzed and subjects were categorized into low, medium and high level groups according to stool Lactobacillus measurements. The primary endpoints were major adverse cardiac events. Cox regression analysis was used to analyze the relationship between Lactobacillus and prognosis. Spearman correlation analysis and trend tests were used to assess the relationship between Lactobacillus and the clinical indicators. RESULTS: The data of 254 patients were included in the analysis. Mean age was 65.90 ± 11.56 years, and 152 patients (59.84%) were male. Follow-up time was 652 (548.25-753.00) days. Multivariate Cox regression analysis showed a significantly lower risk of major adverse cardiac events in patients with Lactobacillus > 7.1 copies/g [adjusted hazard ratio (HR) = 0.216, 95% CI: 0.094-0.493,P < 0.001] compared to patients with Lactobacillus ≤ 3.6 copies/g. Statistically significant differences were shown in ST-segment elevation myocardial infarction (STEMI) (HR = 0.217, 95% CI: 0.085-0.551, P = 0.001). Lactobacillus was a protective factor for male smokers aged over 60 years whose brain natriuretic peptide was over 1,000 pg/mL. Spearman correlation analysis showed that Lactobacillus correlated negatively with white blood cells, neutrophils, high-sensitivity C-reactive protein, TroponinT, creatine kinase, creatine kinase-MB and brain natriuretic peptide (downward trend), and correlated positively with left ventricular ejection fraction (upward trend). CONCLUSIONS: This study is the first to reveal the correlation between Lactobacillus and inflammation and myocardial damage after STEMI. STEMI patients, especially male smokers aged over 60 years with severe impairment of cardiac function, have better outcomes with high levels of Lactobacillus, suggesting new therapeutic strategies for improving the prognosis and quality of life of AMI patients.

Human cerebral organoids establish subcortical projections in the mouse brain after transplantation.

Numerous studies have used human pluripotent stem cell-derived cerebral organoids to elucidate the mystery of human brain development and model neurological diseases in vitro, but the potential for grafted organoid-based therapy in vivo remains unknown. Here, we optimized a culturing protocol capable of efficiently generating small human cerebral organoids. After transplantation into the mouse medial prefrontal cortex, the grafted human cerebral organoids survived and extended projections over 4.5 mm in length to basal brain regions within 1 month. The transplanted cerebral organoids generated human glutamatergic neurons that acquired electrophysiological maturity in the mouse brain. Importantly, the grafted human cerebral organoids functionally integrated into pre-existing neural circuits by forming bidirectional synaptic connections with the mouse host neurons. Furthermore, compared to control mice, the mice transplanted with cerebral organoids showed an increase in freezing time in response to auditory conditioned stimuli, suggesting the potentiation of the startle fear response. Our study showed that subcortical projections can be established by microtransplantation and may provide crucial insights into the therapeutic potential of human cerebral organoids for neurological diseases.

Induction of human somatostatin and parvalbumin neurons by expressing a single transcription factor LIM homeobox 6.

Human GABAergic interneurons (GIN) are implicated in normal brain function and in numerous mental disorders. However, the generation of functional human GIN subtypes from human pluripotent stem cells (hPSCs) has not been established. By expressing LHX6, a transcriptional factor that is critical for GIN development, we induced hPSCs to form GINs, including somatostatin (SST, 29%) and parvalbumin (PV, 21%) neurons. Our RNAseq results also confirmed the alteration of GIN identity with the overexpression of LHX6. Five months after transplantation into the mouse brain, the human GABA precursors generated increased population of SST and PV neurons by overexpressing LHX6. Importantly, the grafted human GINs exhibited functional electrophysiological properties and even fast-spiking-like action potentials. Thus, expression of the single transcription factor LHX6 under our GIN differentiation condition is sufficient to robustly induce human PV and SST subtypes.

Protective effects of seahorse extracts in a rat castration and testosterone-induced benign prostatic hyperplasia model and mouse oligospermatism model.

This study investigated the effects of seahorse (Hippocampus spp.) extracts in a rat model of benign prostatic hyperplasia (BPH) and mouse model of oligospermatism. Compared to the sham operated group, castration and testosterone induced BPH, indicated by increased penile erection latency; decreased penis nitric oxide synthase (NOS) activity; reduced serum acid phosphatase (ACP) activity; increased prostate index; and epithelial thickening, increased glandular perimeter, increased proliferating cell nuclear antigen (PCNA) index and upregulation of basic fibroblast growth factor (bFGF) in the prostate. Seahorse extracts significantly ameliorated the histopathological changes associated with BPH, reduced the latency of penile erection and increased penile NOS activity. Administration of seahorse extracts also reversed epididymal sperm viability and motility in mice treated with cyclophosphamide (CP). Seahorse extracts have potential as a candidate marine drug for treating BPH without inducing the side effects of erectile dysfunction (ED) or oligospermatism associated with the BPH drug finasteride.

Zinc(II)-curcumin accelerates the healing of acetic acid-induced chronic gastric ulcers in rats by decreasing oxidative stress and downregulation of matrix metalloproteinase-9.

Gastric ulcers form as a result of a multifaceted process which includes acid secretion, reactive oxygen species generation and extracellular matrix (ECM) degradation. The aim of this study was to investigate the possible mechanisms underlying the anti-ulcerogenic effects of the Zn(II)-curcumin complex, a curcumin derivative, on the healing of acetic acid-induced gastric ulcers in rats. The severely ulcerated gastric mucosa of control animals had a lower glutathione level (GSH) and superoxide dismutase activity (SOD), and increased malondialdehyde (MDA) content compared to sham operated rats (P<0.001). Zn(II)-curcumin solid dispersions (equivalent to 12, 24 and 48 mg/kg) dose-dependently reduced the gastric ulcer index, significantly increased SOD activity and GSH levels, and reduced the MDA content and matrix metalloproteinase-9 (MMP-9) mRNA expression in the gastric mucosa (P<0.05, compared to control animals). Zn(II)-curcumin exerted a greater anti-ulcerogenic effect than curcumin at the same dose (24 mg/kg), leading to a reduced severity of gastric ulcers, lower MDA content, and increased SOD activity and GSH levels (P<0.05). In conclusion, these results confirm that the Zn(II)-curcumin complex possesses an enhanced mucosal barrier defense activity compared to curcumin alone, due to its synergistic ability to decrease oxidative stress and attenuate MMP-9-mediated inflammation.

[Studies on solubility enhancement of curcumin by Polyvinylpyrrolidione K30].

OBJECTIVE: To study solubility enhancement of curcumin by Polyvinylpyrrolidione K30 (PVP K30). METHODS: Solid dispersion systems (SDS) of curcumin in PVP K30 were prepared at various weight ratios by co-evaporation of curcumin and PVP K30 ethanol solution. The differential scanning calorimetry (DSC) and powder X-ray diffractometer method were used to describe the status of curcumin in carriers, the UV spectrometry method for determination of curcumin in mediums was established. RESULTS: The curcumin SDS was successfully prepared, the UV spectrometry method was accurate and reliable, and no interference occurred from carrier. The solubility rate in vitro of curcumin was significantly raised. Compared to curcumin, the solubility of curcumin in SDS increased at least 880 folds. CONCLUSION: PVP K30 improves the solubility of curcumin well.

[Effects of alligator Zhikegao on relieving cough dispelling phlegm and anti-inflammation].

OBJECTIVE: To research the effects of Alligator Zhikegao on relieving cough, dispelling phlegm and anti-inflammation. METHOD: The coughing tests in mice, the phenol red secreting tests in mice, ear edema tests in mice,and paw edema tests and subcutaneous cotton ball granuloma in rats were adopted for observing the related pharmacological effects of Alligator Zhikegao. RESULT: Alligator Zhikegao could obviously prolong the latent period and decrease the times of mouse coughing, and remarkably inhibit the mouse ear edema (P < 0.001), the rat paw edema and the hyperplasia of subcutaneous cotton ball granuloma in rats. Alligator Zhikegao 11.70 g x kg(-1) could significant improve the carbonic clearances of macrophages (P <0.05) and the hemolysin level in serum (P <0.01). CONCLUSION: Alligator Zhikegao has significant effects on relieving cough, dispelling phlegm, anti-inflammation and immunological regulation.

Determination of a new active steroid by high performance liquid chromatography with laser-induced fluorescence detection following the pre-column derivatization.

A sensitive analytical method for the determination of a new active steroid, butane acid-(5-androsten-17-one-3beta-ol)-diester (A1998), was developed by high performance liquid chromatography with laser-induced fluorescence detection following the pre-column derivatization with dansylhydrazine. The calibration curve for A1998 derivatization was found linear in the dynamic range from 0.025 to 5.0 microg/ml, with the precision less than 6% (CV) and the mean extraction efficiency greater than 92%. In 200 microl of plasma samples the limit of quantitation was as low as 0.025 microg/ml with a signal-to-noise ratio of 10. This assaying was further applied to the determination of the pharmacokinetic parameters of A1998 in rats with an intravenous injection of A1998. Values for clearance for elimination, volume of distribution at steady state and terminal half life in the above case were determined as 50.3+/-1.1 ml/min kg, 1329.0+/-111.0 ml/kg and 44.0+/-2.7 min, respectively.

Protective effects of 5,4'-dihydroxy-3',5'-dimethoxy-7-O-beta-D -glucopyranosyloxy-flavone on experimental hepatic injury.

AIM: To investigate the pharmacological effects of rice flavone (5,4'-dihydroxy-3',5'-dimethoxy-7-O-beta-D-glucopyranosyloxy-flavone, RF) separated from panicle-differentiating to flowing rice on rat experimental hepatic injury. METHODS: Models of rat acute hepatic injury induced by carbon tetrachloride (CCl(4)) administration, rat hepatic fibrosis induced by thioacetamide, injury of primary cultured rat hepatocytes induced by CCl(4), respectively, were established. After treated with RF, content of serum alanine transaminase (ALT), aspartate aminotransferase (AST) and albumin (Alb), hyaluronic acid (HA), the activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and hydroxyproline (Hyp) were measured and liver tissue was observed pathologically by hematoxylin-eosin (HE) staining. Effects of RF on pathological changes, function index, enzyme of scavenging free radicals and blood rheology were evaluated. RESULTS: In model of rat acute hepatic injury induced by CCl(4), RF can significantly decrease the contents of serum ALT, AST, increase the content of Alb, improve the dropsy and fat denaturalization of hepatocytes. In model of rat hepatic fibrosis induced by thioacetamide, RF can inhibit the increase of HA, Hyp and whole blood viscosity, and improve the activities of GSH-Px and SOD, and inauricular microcirculation. CONCLUSION: RF has apparent protective effects on hepatic injury by increasing activity of GSH-Px and SOD, scavenging free radicals produced by CCl(4), reducing blood viscosity, and improving microcirculation and blood supply.

[Protective effects of polysacchride of Spirulina platensis and Sargassum thunbeergii on vascular of alloxan induced diabetic rats].

OBJECTIVE: To study the protective effects of polysaccharide of Spirulina platensis and Sargassum thunbeergii on vascular of alloxan (ALX) induced diabetic rats. METHOD: With the doses of polysaccharide of Spirulina platensis (PSP) and Sargassum thunbeergii (PST) compound (1:1) 12.261, 36.783, 110.349 mg x kg(-1) by i.g. administration to alloxan induced diabetic rats respectively for 6 weeks. Then the blood glucose and the TC, HDL-C, TG, NO, ET in serum were detected. The contraction and relaxation response to NE and ACh in aortic rings of the alloxan induced diabetic rats has been studied. RESULT: The results showed the compound of PSP and PST could decrease the blood glucose and the TC, TG, NO, ET in serum and increase HDL-C than in the alloxan induced diabetic rats. The contraction responses to NE in aortic rings of the alloxan induced diabetic rats were significantly elevated in the normal rats, and the responses to ACh were significantly lower. PSP and PST compound could significantly lower the responses to NE and significantly elevate the responses to ACh in aortic rings of the alloxan induced diabetic rats. CONCLUSION: PSP and PST compound could decrease blood glucose and could protect the vascular of alloxan induced diabetic rats.

[Preparation and evaluation in vivo and in vitro of glimepiride gel-matrix controlled-release patch].

AIM: To study the pharmaceutical characterization, the pharmacokinetics and relative bioavailability of glimepiride gel-matrix controlled-release patch in rats. METHODS: An HPLC method was established for the determination of glimepiride in the permeation receptor and patch. The permeation rate and penetration mechanism of glimepiride-TDDS through rabbit skin in vitro was examined. The determination of drug content and the examination of weight difference and stability of the glimepiride-TDDS were carried out. Another HPLC method after pre-column derivatization was developed to determine the glimepiride serum concentration and then employed to study the pharmacokinetics and relative bioavailability of glimepiride after a single dose of oral or patch administration in rats. RESULTS: The permeation tests through excised rabbit skin demonstrated that the optimized glimepiride controlled-release patch exhibited zero-order kinetic characteristics that satisfied the demands of original design. The determination of glimepiride content and the quality control of weight difference of the patch accorded with Pharmacopoeia of the People's Republic of China of 2000 edition and the pharmaceutical characterization showed good stability. The HPLC method for the determination of serum glimepiride was shown to be a sensitive and simple one. The pharmacokinetic results showed that TDDS could decrease the maximum serum concentration, prolong the peak time, extend the MRT by 5.5 times compared with oral administration and maintain the serum concentration of glimepiride at a higher level even after 120 h of administration. The relative bioavailability of glimepiride-TDDS was 20.3% versus oral administration. CONCLUSION: The glimepiride-TDDS showed a slower, longer and smoother serum concentration-time profile, as compared with conventional oral administration in both absorption and elimination phase. As a result, it was evident that the patch exhibited good controlled-release properties.

[Anti-arrhythmic effect of starfish sterol].

AIM: To study the effect of modified starfish sterol [C03, succinic acid (5-epiandroene-17-one-3beta-ol) diester] on experimental arrhythmias. METHODS: Arrhythmias were induced by drugs (Aco, Oua, BaCl2 and adrenalin) i.v., ligating the left anterior descending coronary artery and electricity. RESULTS: C03 71.4 mg x kg(-1) (ig) was shown to increase the dose of Oua inducing VP, VT, VF and CA in guinea pigs (P < 0.01); C03 (26.8, 80.4 mg x kg(-1)) was found to increase the dose of Aco inducing VF and CA in rats (P < 0.01); C03 (8.9, 26.8, 80.4 mg x kg(-1)) increase the dose of barium chloride and delay the onset time of ventricular arrhythmias (P < 0.01); C03 (14.1, 42.3 mg x kg(-10) shorten time of recovering induced by adrenalin in rabbits (P < 0.01); C03 (80.4 mg x kg(-1)) was shown to reduce the number of ventricular arrhythmias induced by coronary artery ligation in rats (P < 0.05), C03 increase VFT induced by electricity in rabbits, VFT of C03 14.1 mg x kg(-1) increased from (5.1 +/- 2.5) V to (11.0 +/- 2.7) V (P < 0.01), 42.3 mg x kg(-1) increased from (6.1 +/- 1.7) V to (15 +/- 5) V (P < 0.01). CONCLUSION: Starfish sterol has anti-arrhythmic effect.

[The antitussive and de-sputum essential substance from the root of Gerbera piloselloides and its analytical compositions].

OBJECTIVE: To study the chemical constituents of the essential substance from the root of Gerbera piloselloides and its antitussive and de-sputum effects. METHOD: The essential substance (G4) was extracted from the root by alcohol and ethyl acetate, then it was separated by silica gel column eluted by the mixture of ethyl acetate and petroleum ether (5:95). Its chemical components were separated and identified by GC-MS. Its antitussive and de-sputum effect was tested by mice. RESULT: 4 main peaks were separated and identified by GS-MS. They are beta-caryophyllene (15.160%), caryophyllene oxide (21.140%), aristolenepoxide (2.673%) and 6-acetyl-2,2-dimethyl-8(3-methyl-2-butenyl)-2H-chromoene (60.077%) respectively. Its antitussive and de-sputum effect was prominent when the mice was given G4 2,000 mg.kg-1 ig. CONCLUSION: Itis the first time that the antitussive and de-sputum essential substance was separated from the root of Gerbera piloselloides and its main compositions were analyzed.