ObjectiveTo evaluate the effects of Tongnaoyin on the blood-brain barrier status and neurological impairment in acute ischemic stroke （AIS） patients with the syndrome of phlegm-stasis blocking collaterals by dynamic contrast-enhanced magnetic resonance imaging （DCE-MRI）. MethodsA total of 63 patients diagnosed with AIS in the Jiangsu Province Hospital of Chinese Medicine from October 2022 to December 2023 were enrolled in this study. According to random number table method，the patients were assigned into a control group （32 cases） and an observation group （31 cases）. The control group received conventional Western medical treatment，and the observation group took 200 mL Tongnaoyin after meals，twice a day from day 2 of admission on the basis of the treatment in the control group. After 7 days of treatment，the patients were examined by DCE-MRI. The baseline data for two groups of patients before treatment were compared. The National Institute of Health Stroke Scale （NIHSS） score and modified Rankin Scale （mRS） score were recorded before treatment and after 90 days of treatment for both groups. The rKtrans，rKep，and rVe values were obtained from the region of interest （ROI） of the infarct zone/mirror area and compared between the two groups. ResultsThere was no significant difference in the NIHSS or mRS score between the two groups before treatment. After 90 days of treatment，the NIHSS and mRS scores declined in both groups，and the observation group had lower scores than the control group （P<0.05）. After treatment，the rKtrans and rVe in the observation group were lower than those in the control group （P<0.01）. ConclusionCompared with conventional Western medical treatment alone，conventional Western medical treatment combined with Tongnaoyin accelerates the repair of the blood-brain barrier in AIS patients，thereby ameliorating neurological impairment after AIS to improve the prognosis.

ObjectiveTo investigate the changes in cerebral blood perfusion in patients with acute cerebral infarction after taking Tongnaoyin， a traditional Chinese medicine， based on head and neck computed tomography （CT） angiography （CTA） combined with brain CT perfusion imaging （CTP）. MethodA total of 240 patients with cerebral infarction of phlegm and blood stasis syndrome treated in Jiangsu Province Hospital of Traditional Chinese Medicine from March 2018 to September 2023 were randomly divided into a control group （99 cases） and a Tongnaoyin group （141 cases）. Based on the guidelines， the control group was treated with conventional treatment such as anti-aggregation， anticoagulation， lipid-lowering and plaque stabilization， brain protection， and supportive treatment. The Tongnaoyin group was treated with Tongnaoyin of 200 mL in warm conditions in the morning and evening on the basis of the control group. Both groups underwent CTA combined with CTP within 24 hours after admission， and they were reexamined by CTA and CTP in the sixth month after admission. The degree of intracranial artery stenosis was determined according to the North American Symptomatic Carotid Endarterectomy Trial （NASCET） method. The relative cerebral blood volume （rCBV）， relative cerebral blood flow （rCBF）， mean transit time （MTT）， and time to peak （TTP） of the lesion area before and after treatment were compared. The adverse outcomes of the two groups within six months after discharge were compared. ResultCompared with the group before treatment， the degree of vascular stenosis in the Tongnaoyin group was significantly reduced， and the difference was statistically significant （Z=105.369，P<0.05）. Compared with the control group after treatment， the improvement rate of vascular stenosis in the Tongnaoyin group was higher （χ²=84.179，P<0.01）， and the curative effect was better.After treatment， the rCBV and rCBF of patients in the Tongnaoyin group were significantly increased， and the difference was statistically significant （P<0.01）. MTT and TTP showed a trend of shortening， but the difference was not statistically significant. There was no statistically significant difference in rCBV， rCBF， MTT， and TTP in the control group. Compared with those in the control group after treatment， the rCBV and rCBF in the Tongnaoyin group were significantly increased， while MTT and TTP were significantly reduced （P<0.01）. After six months of discharge， the risk of poor prognosis in the Tongnaoyin group was significantly reduced compared with the control group （P<0.05）. ConclusionTongnaoyin has a good effect on improving cerebral blood perfusion in patients with acute cerebral infarction. It can be used as an effective supplement for the conventional treatment of ischemic stroke to improve clinical efficacy.

Aim To explore the mechanism of oxiracetam promoting neurogenesis and migration in rats with cer-ebral infarction through stromal cell-derived factor-1α(SDF-1α)/C-X-C chemokine receptor 4(CXCR4)pathway.Methods 100 SD rats were randomly divided into control group,cerebral ischemia(CI)group,oxiracetam(200 mg/kg)group,and oxiracetam(200 mg/kg)+AMD3100(5 mg/kg)group,with 25 rats in each group.Electrocoagulation was used to create rat model of local permanent cerebral infarction.After 1,7 and 14 days of modeling,neurological deficits were scored,TTC staining was used to detect the volume of cerebral infarction,Nissl staining was used to detect cell surviv-al in the infarcted area,Western blot was used to detect SDF-1α and CXCR4 protein levels in ischemic zone.After 1～7 days of modeling,BrdU(50 mg/kg)was continuously injected intraperitoneally.After 14 days,immunofluorescence double staining was used to detect the number of BrdU+Nestin+and BrdU+DCX+cells in the SVZ region.5 days before modeling,retroviruses carrying GFP were injected into the SVZ region.After 14 days,immunofluorescence double stai-ning was used to detect the number of GFP+DCX+,GFP+MAP-2+and GFP+GFAP+cells in infarction area.C17.2 cells were divided into control group,oxygen-glucose deprivation(OGD)group,oxiracetam(final concentration:200 mg/L)group,and oxiracetam(final concentration:200 mg/L)+AMD3100(final concentration:100 μmol/L)group.OGD was used to create cell CI model.After 12 hours,immunofluorescence double staining was used to detect the number of Br-dU+/Nestin+and BrdU+/MAP-2+cells,Transwell experiment was used to detect cell migration,Western blot was used to detect SDF-1α and CXCR4 protein levels in cell culture supernatant.Results Animal experiment results showed:compared with control group,mNSS score in CI group was increased,cerebral infarction volume was increased,the number of surviving cells in infarcted area was decreased,SDF-1α and CXCR4 protein levels were increased,the number of GFP+DCX+,GFP+MAP-2+and GFP+GFAP+cells in SVZ region were increased(P＜0.05);compared with CI group,mNSS score in oxiracetam group was decreased,cerebral infarction volume was decreased,the number of surviving cells in infarc-ted area was increased,SDF-1α and CXCR4 protein levels were increased,the number of GFP+DCX+,GFP+MAP-2+and GFP+GFAP+cells in SVZ region were increased,the number of GFP+DCX+,GFP+MAP-2+and GFP+GFAP+cells in in-farcted area were increased(P＜0.05);compared with oxiracetam group,mNSS score in oxiracetam+AMD3100 group was increased,cerebral infarction volume was increased,the number of surviving cells in infarcted area was decreased,CXCR4 protein level was decreased,the number of GFP+DCX+,GFP+MAP-2+and GFP+GFAP+cells in the SVZ region were de-creased(P＜0.05).Cell experiment results showed:compared with control group,the number of BrdU+/Nestin+and Br-dU+/MAP-2+cells in OGD group were increased,the number of cell migration,SDF-1α and CXCR4 protein levels in cell culture supernatant were increased(P＜0.05);compared with OGD group,the number of BrdU+/Nestin+and BrdU+/MAP-2+cells in oxiracetam group were increased,the number of cell migration,SDF-1α and CXCR4 protein levels in cell culture supernatant were increased(P＜0.05);compared with oxiracetam group,the number of BrdU+/Nestin+and BrdU+/MAP-2+cells in oxiracetam+AMD3100 group were decreased,the number of cell migration,CXCR4 protein level in cell culture supernatant were decreased(P＜0.05).Conclusion Oxiracetam may promote the migration of neural stem cells from the SVZ region to the ischemic zone,promoting neurogenesis and functional recovery in rats with cerebral infarction by activating SDF-1α/CXCR4 pathway.

Objective:To investigate the effect of empagliflozin on diabetic kidney disease in db/db mice and the potential mechanisms.Methods:db/db mice were randomly divided into db/db group and Empa group. C57BL/6J mice were used as normal control group. We measured the level of serum biochemistry and inflammatory cytokines. Pathological changes of kidney were observed by pathological staining. The protein expression levels of NLRP3, cleaved caspase-1 and GSDMD were detected.Results:Compared with db/db group, the level of fasting blood glucose, blood lipids, serum biochemistry, and urinary protein-to-creatinine ratio in Empa group were significantly decreased ( P<0.05). HE staining and Masson staining showed that empagliflozin could significantly improve glomerular pyknosis and renal interstitial fibrosis in db/db mice. Meanwhile, the expressions of NLRP3, cleaved caspase-1, and GSDMD protein were down-regulated ( P<0.05). Conclusion:Empagliflozin improves kidney damage in diabetic model mice, and the possible mechanism is to inhibit the cell pyroptosis signal pathway of NLRP3/caspase-1/GSDMD.

COVID-19 has swept globally and Pakistan is no exception.To investigate the initial introductions and transmissions of the SARS-CoV-2 in Pakistan,we performed the largest genomic epidemiology study of COVID-19 in Pakistan and generated 150 complete SARS-CoV-2 genome sequences from samples collected from March 16 to June 1,2020.We identified a total of 347 mutated positions,31 of which were over-represented in Pakistan.Meanwhile,we found over 1000 intra-host single-nucleotide variants(iSNVs).Several of them occurred concurrently,indicating possible interactions among them or coevolution.Some of the high-frequency iSNVs in Pakistan were not observed in the global population,suggesting strong purifying selections.The genomic epidemiology revealed five distinctive spreading clusters.The largest cluster consisted of 74 viruses which were derived from different geographic locations of Pakistan and formed a deep hierarchical structure,indicating an extensive and persistent nation-wide transmission of the virus that was probably attributed to a signature mutation(G8371T in ORF 1ab)of this cluster.Further-more,28 putative international introductions were identified,several of which are consistent with the epidemiological investigations.In all,this study has inferred the possible pathways of introduc-tions and transmissions of SARS-CoV-2 in Pakistan,which could aid ongoing and future viral surveillance and COVID-19 control.

Objective:To investigate the expression of costimulatory molecule OX40 in peripheral CD4 +CD25 +CD127 low regulatory T (Treg) cells and its clinical significance in patients with acute cerebral infarction (ACI). Methods:Seventy-five patients with first-onset ACI, admitted to our hospital from April 2019 to December 2019, and 36 age- and gender-matched volunteers (control group) were selected in this study. OX40 expression on CD4 +CD25 +CD127 low Treg cells in peripheral blood samples in the two groups were analyzed by immunofluorescent labeling and flow cytometry. Correlations of OX40 +Treg cell percentage with National Institute of Health stroke scale (NIHSS) scores, ischemic penumbra volume, core infarct volume, and infarct volume in the patient group were analyzed. The changes of OX40 +Treg cell percentage in the patient group before and after endovascular treatment or intravenous thrombolysis were compared. Results:As compared with that in the control group, the Treg cell percentage in peripheral blood samples of the patient group was significantly decreased, while OX40 +Treg cell percentage was significantly increased ( P<0.05). OX40 +Treg cell percentage was positively correlated with NIHSS scores in ACI patients ( r s=0.271, P=0.018). Meanwhile, OX40 +Treg percentage was significantly correlated with ischemic penumbra volume, core infarct volume, and infarct volume in the patient group ( r s=0.435, P=0.000; r s=0.343, P=0.003; r s=0.245, P=0.034). OX40 +Treg cell percentage in ACI patients 7 d after endovascular treatment was significantly decreased as compared with that before treatment ( P<0.05); OX40 +Treg percentage in ACI patients 3 and 7 d after intravenous thrombolysis was significantly decreased as compared with that before treatment ( P<0.05). Conclusion:OX40 is abnormally expressed on peripheral Treg cells in ACI patients, and is closely correlated with neurological deficits, imaging features and reperfusion therapy.

On January 22, 2020, China National Center for Bioinformation (CNCB) released the 2019 Novel Coronavirus Resource (2019nCoVR), an open-access information resource for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). 2019nCoVR features a comprehensive integra-tion of sequence and clinical information for all publicly available SARS-CoV-2 isolates, which are manually curated with value-added annotations and quality evaluated by an automated in-house pipeline. Of particular note, 2019nCoVR offers systematic analyses to generate a dynamic landscape of SARS-CoV-2 genomic variations at a global scale. It provides all identified variants and their detailed statistics for each virus isolate, and congregates the quality score, functional annotation,and population frequency for each variant. Spatiotemporal change for each variant can be visualized and historical viral haplotype network maps for the course of the outbreak are also generated based on all complete and high-quality genomes available. Moreover, 2019nCoVR provides a full collection of SARS-CoV-2 relevant literature on the coronavirus disease 2019 (COVID-19), including published papers from PubMed as well as preprints from services such as bioRxiv and medRxiv through Europe PMC. Furthermore, by linking with relevant databases in CNCB, 2019nCoVR offers data submission services for raw sequence reads and assembled genomes, and data sharing with NCBI. Collectively, SARS-CoV-2 is updated daily to collect the latest information on genome sequences, variants, hap-lotypes, and literature for a timely reflection, making 2019nCoVR a valuable resource for the global research community. 2019nCoVR is accessible at https://bigd.big.ac.cn/ncov/.

Objective: To prepare manganese-doped carbon quantum dots (Mn-CDs) dual-modal nanoprobe for fluorescent-magnetic imaging, and evaluate its characteristics and potential on fluorescence imaging and MRI. Methods: Mn-CDs were synthesized at 150 ℃. The form, diameter, component, fluorescent capability, T₁ relaxation rate, stability and cytotoxicity of Mn-CDs in vivo were verified. The fluorescence imaging of HO-8910 tumor-bearing mice was performed on small animal imager, and the whole-body enhanced imaging was performed on 3.0 T MRI scanner. One-way analysis of variance was used to analyze the data. Results: The Mn-CDs with the diameter of (4.64±0.85) nm showed a well-defined spherical morphology. The fluorescent spectra of Mn-CDs exhibited that the excitation maximum was at 360 nm and the emission maximum was at 440 nm. The T₁ relaxation rate was (3.26±0.04) mmol·L^-1·s^-1. The Mn-CDs had good stability of fluorescent and magnetic imaging capability at 0, 0.25, 0.5, 0.75, 1.0 and 2 months at room temperature with no significant differences of fluorescent and magnetic signals (F=1.566 and 0.987, both P>0.05). After injection of 200 μl Mn-CDs (15 g/L), mice were all alive and had no viscera damage. The tumor could be observed obviously on fluorescence imaging at 5 min. Enhanced MRI showed that the tumor was unevenly enhanced and Mn-CDs were mainly cleared away through urinary system. Conclusion Mn-CDs are stable and have good potential on fluorescence imaging and MRI, which provides a promising multimodal imaging method for tumor detection and monitoring.

Objective To explore the expressions of costimulatory molecules OX40 and OX40L in peripheral blood (PB) samples of multiple sclerosis (MS) patients,and reveal their clinical significance.Methods PB samples of 39 MS patients and 37 health control subjects (HC) were collected from June 2013 to October 2018.OX40 expressions on CD4+ T cells,OX40L expressions on CD14+ monocytes and CD19+ B cells were detected by immunofluorescent labeling and flow cytometer.The correlations between expanded disability status scale (EDSS) scores and OX40 and OX40L expressions were analyzed,and the influences of different clinical typing and gender in OX40 and OX40L expressions were analyzed.The changes of OX40 and OX40L expressions before and after methylprednisolone shock therapy in 8 patients with recurrent MS were observed.Results (1) As compared with those in the HC group,the expressions of CD4+OX40+ and CD 14+OX40L+ in the peripheral blood of patients in the MS group were significantly increased (P＜0.05).(2) The OX40 expression on the surface of CD4+ T cells was positively correlated with EDSS scores of MS patients (r=0.684,P=0.000).(3) As compared with that in the MS patients at remission phase,the OX40 expression in the CD4+ T cells of MS patients at recurrence phase was significantly increased (P＜0.05);there were no significant differences in the expressions of OX40 and OX40L between male and female MS patients (P＞0.05).(4) EDSS scores of 8 patients with relapsed MS after intravenous infusion of methylprednisolone were significantly lower than those before treatment (P＜0.05);the OX40 expression on the surface of CD4+ T cells after intravenous infusion of methylprednisolone was significantly decreased as compared with that before treatment (P＜0.05).Conclusion Abnormal OX40 expression is closely correlated to neurologic impairment and clinical characteristics,and distinctly responded to corticosteroid therapy in MS patients,which suggests that OX40 may be a promising biomarker and is involved in the pathogenesis of MS.

Objective To investigate the relationship between abnormal bone mineral density (BMD) and subclinical thyroid dysfunction. Methods Thyroid function, biochemical indicators of bone metabolism and BMD were reviewed retrospectively in the subjects who received health checkups from July 1, 2009 to January 31, 2017 in the Health Check-up Department of Peking Union Medical College Hospital. People who had thyroid dysfunction, recognized risk factors for osteoporosis, and medication history were excluded. A cross-sectional analysis of thyroid status and biochemical indicators of bone metabolism was performed by the standard methods. BMD at the lumbar spine and femoral neck was measured using dual energy X-ray absorptiometry. Results A total of 6884 subjects (3726 women and 3158 men) were enrolled in the study, with an average age of (50.74 ± 10.41) years. They were divided into three groups:subclinical hyperthyroid, subclinical hypothyroid, and euthyroidism. The alkaline phosphatase in subclinical hyperthyroid group was higher than that in the euthyroidism group[ (67.95±20.64)U/L vs. (63.88±18.99)U/L]. Calcium and phosphorus in blood were higher in both subclinical hyperthyroid and subclinical hypothyroid groups. The rate of abnormal BMD in male euthyroidism, subclinical hyperthyroid and subclinical hypothyroid groups were 36.10％(1049/2906), 29.27％(12/41) and 27.01％(57/211), respectively. The rate of abnormal BMD showed no difference between subclinical hyperthyroid group and euthyroidism group (P>0.05). The rate of abnormal BMD was lower in subclinical hypothyroid group than in euthyroidism group (χ2=7.0901, P<0.01), especially in the males aged 40-49 years (χ2=10.4618, P<0.01). The rate of abnormal BMD in female euthyroidism, subclinical hyperthyroid and subclinical hypothyroid groups was 38.81％(1286/3314), 45.83％ (33/72) and 40.88％ (139/340), respectively. The rate of abnormal BMD showed no difference among the three groups (P>0.05). Conclusion There is no significant difference in the rate of abnormal BMD between subclinical thyroid dysfunction group and euthyroidism group, possibly because abnormal serum biochemical indicators preceded the presence of low BMD. More sensitive methods used to determine the status of bone metabolism await to be developed.