ObjectiveTo observe the clinical efficacy and safety of Tangning Tongluo tablets in the treatment of nonproliferative diabetic retinopathy （DR）. MethodsFourteen research centers participated in this study， which spanned a time interval from September 2021 to May 2023. A total of 240 patients with nonproliferative DR were included and randomly assigned into an observation group （120 cases） and a control group （120 cases）. The observation group was treated with Tangning Tongluo tablets， and the control group with calcium dobesilate capsules. Both groups were treated for 24 consecutive weeks. The vision， DR progression rate， retinal microhemangioma， hemorrhage area， exudation area， glycosylated hemoglobin （HbA1c） level， and TCM syndrome score were assessed before and after treatment， and the safety was observed. ResultsThe vision changed in both groups after treatment （P<0.05）， and the observation group showed higher best corrected visual acuity （BCVA） than the control group （P<0.05）. The DR progression was slow with similar rates in the two groups. The fundus hemorrhage area and exudation area did not change significantly after treatment in both groups， while the observation group outperformed the control group in reducing the fundus hemorrhage area and exudation area. There was no significant difference in the number of microhemangiomas between the two groups before treatment. After treatment， the number of microhemangiomas decreased in both the observation group （Z=-1.437， P<0.05） and the control group （Z=-2.238， P<0.05）， and it showed no significant difference between the two groups. As the treatment time prolonged， the number of microhemangiomas gradually decreased in both groups. There was no significant difference in the HbA1c level between the two groups before treatment. After treatment， the decline in the HbA1c level showed no significant difference between the two groups. The TCM syndrome score did not have a statistically significant difference between the two groups before treatment. After treatment， neither the TCM syndrome score nor the response rate had significant difference between the two groups. With the extension of the treatment time， both groups showed amelioration of TCM syndrome compared with the baseline. ConclusionTangning Tongluo tablets are safe and effective in the treatment of nonproliferative DR， being capable of improving vision and reducing hemorrhage and exudation in the fundus.

Guided by the concept of quality by design(QbD), this study optimizes the calcination and quenching process of calcined Magnetitum and establishes the XRD characteristic spectra of calcined Magnetitum, providing a scientific basis for the formulation of quality standards. Based on the processing methods and quality requirements of Magnetitum in the Chinese Pharmacopoeia, the critical process parameters(CPPs) identified were calcination temperature, calcination time, particle size, laying thickness, and the number of vinegar quenching cycles. The critical quality attributes(CQAs) included Fe mass fraction, Fe~(2+) dissolution, and surface color. The weight coefficients were determined by combining Analytic Hierarchy Process(AHP) and the criteria importance though intercrieria correlation(CRITIC) method, and the calcination process was optimized using orthogonal experimentation. Surface color was selected as a CQA, and based on the principle of color value, the surface color of calcined Magnetitum was objectively quantified. The vinegar quenching process was then optimized to determine the best processing conditions. X-ray diffraction(XRD) was used to establish the characteristic spectra of calcined Magnetitum, and methods such as similarity evaluation, cluster analysis, and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to evaluate the quality of the spectra. The optimized calcined Magnetitum preparation process was found to be calcination at 750 ℃ for 1 h, with a laying thickness of 4 cm, a particle size of 0.4-0.8 cm, and one vinegar quenching cycle(Magnetitum-vinegar ratio 10∶3), which was stable and feasible. The XRD characteristic spectra analysis method, featuring 9 common peaks as fingerprint information, was established. The average correlation coefficient ranged from 0.839 5-0.988 1, and the average angle cosine ranged from 0.914 4 to 0.995 6, indicating good similarity. Cluster analysis results showed that Magnetitum and calcined Magnetitum could be grouped together, with similar compositions. OPLS-DA discriminant analysis identified three key characteristic peaks, with Fe_2O_3 being the distinguishing component between the two. The final optimized processing method is stable and feasible, and the XRD characteristic spectra of calcined Magnetitum was initially established, providing a reference for subsequent quality control and the formulation of quality standards for calcined Magnetitum.
X-Ray Diffraction/methods* ; Drugs, Chinese Herbal/chemistry* ; Quality Control ; Particle Size

Objective To investigate the weight loss efficacy and mechanisms of oligopeptide-herbal medicine composite,and to provide new approaches for obesity treatment.Methods Twenty-three SPF female SD rats were randomly divided into control group(n=3)and modeling group(n=20).The control group was fed an ordinary diet for 6 consecutive weeks,and the modeling group was fed a high-fat diet for 6 consecutive weeks to establish a simple obesity rat model.After successful modeling,the modeling group was randomly divided into model group(n=10)and treatment group(n=10).The treatment group started anoint-and-massage therapy with oligopeptide-herbal medicine composite(3 g per rat per time,once a day,20 min each time),which was recorded as day 1.The control group and model group were not treated with oligopeptide-herbal medicine composite but received the same massage,and continued to be fed ordinary feed and high-fat feed respectively.Ten rats in the model group and ten in the treatment group were each divided into 3 groups,with 3,3,and 4 rats in each group.On days 3,6,and 9 after treatment,the rats in the three groups were weighed and cardiac blood collection was performed after isoflurane respiratory anesthesia.After euthanasia by cardiac bloodletting,abdominal subcutaneous adipose tissue(aSAT)and perirenal white adipose tissue(pWAT)samples were collected and weighed.Serum triglycerides(TG)and high-density lipoprotein cholesterol(HDL-C)levels were measured.Image J software was used to measure aSAT thickness and the diameter and area of perirenal white adipocytes and abdominal subcutaneous adipocytes.Immunofluorescence technique was used to observe the number of telocytes(TCs),cell junctions and exosomes per unit area of aSAT.Transmission electron microscopy was used to measure the length of telopodes(Tps)of dermal and subcutaneous TCs in abdominal skin,and to observe the distribution of exosome vesicles,rough endoplasmic reticulum,mitochondria,and cell junctions.Results Compared with the control group,the body weight of rats in the modeling group increased significantly(P＜0.05).Compared with the control group,rats in the model group showed a significant increase in serum TG level,a significant decrease in HDL-C level,and a significant increase in pWAT mass(all P＜0.05).Compared with the model group,the treatment group showed a significantly increased rate of body weight reduction,significantly decreased pWAT mass,significantly decreased serum TG level,and significantly increased HDL-C level(all P＜0.05).Compared with the model group,the treatment group showed significant reductions in aSAT thickness,as well as cell diameter and area in both aSAT and pWAT(all P＜0.05).Compared with the model group,the treatment group showed significantly increased number of TCs per unit area of aSAT,number of exosomes from TCs in aSAT,and Tps length(all P＜0.05).The treatment group showed an increasing trend in the numbers of mitochondria,rough endoplasmic reticulum,and cell junctions in TCs.Conclusion Oligopeptide-herbal medicine composite applied via anoint-and-massage therapy effectively improves obesity-related symptoms such as dyslipidemia and fat accumulation in obese rats by regulating TCs and their intercellular communication.

Objective To investigate the weight loss efficacy and mechanisms of oligopeptide-herbal medicine composite,and to provide new approaches for obesity treatment.Methods Twenty-three SPF female SD rats were randomly divided into control group(n=3)and modeling group(n=20).The control group was fed an ordinary diet for 6 consecutive weeks,and the modeling group was fed a high-fat diet for 6 consecutive weeks to establish a simple obesity rat model.After successful modeling,the modeling group was randomly divided into model group(n=10)and treatment group(n=10).The treatment group started anoint-and-massage therapy with oligopeptide-herbal medicine composite(3 g per rat per time,once a day,20 min each time),which was recorded as day 1.The control group and model group were not treated with oligopeptide-herbal medicine composite but received the same massage,and continued to be fed ordinary feed and high-fat feed respectively.Ten rats in the model group and ten in the treatment group were each divided into 3 groups,with 3,3,and 4 rats in each group.On days 3,6,and 9 after treatment,the rats in the three groups were weighed and cardiac blood collection was performed after isoflurane respiratory anesthesia.After euthanasia by cardiac bloodletting,abdominal subcutaneous adipose tissue(aSAT)and perirenal white adipose tissue(pWAT)samples were collected and weighed.Serum triglycerides(TG)and high-density lipoprotein cholesterol(HDL-C)levels were measured.Image J software was used to measure aSAT thickness and the diameter and area of perirenal white adipocytes and abdominal subcutaneous adipocytes.Immunofluorescence technique was used to observe the number of telocytes(TCs),cell junctions and exosomes per unit area of aSAT.Transmission electron microscopy was used to measure the length of telopodes(Tps)of dermal and subcutaneous TCs in abdominal skin,and to observe the distribution of exosome vesicles,rough endoplasmic reticulum,mitochondria,and cell junctions.Results Compared with the control group,the body weight of rats in the modeling group increased significantly(P＜0.05).Compared with the control group,rats in the model group showed a significant increase in serum TG level,a significant decrease in HDL-C level,and a significant increase in pWAT mass(all P＜0.05).Compared with the model group,the treatment group showed a significantly increased rate of body weight reduction,significantly decreased pWAT mass,significantly decreased serum TG level,and significantly increased HDL-C level(all P＜0.05).Compared with the model group,the treatment group showed significant reductions in aSAT thickness,as well as cell diameter and area in both aSAT and pWAT(all P＜0.05).Compared with the model group,the treatment group showed significantly increased number of TCs per unit area of aSAT,number of exosomes from TCs in aSAT,and Tps length(all P＜0.05).The treatment group showed an increasing trend in the numbers of mitochondria,rough endoplasmic reticulum,and cell junctions in TCs.Conclusion Oligopeptide-herbal medicine composite applied via anoint-and-massage therapy effectively improves obesity-related symptoms such as dyslipidemia and fat accumulation in obese rats by regulating TCs and their intercellular communication.

The anti-inflammatory efficacy of Callicarpa nudiflora extract were evaluated upon lipopolysaccharide (LPS)-induced infective inflammation in rats. Pathological changes of lung and colon tissues observed with hematoxylin-eosin (H&E) staining, together with inflammatory factors in serum, including nitric oxide (NO), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), were applied to assess the mitigating effects of C. nudiflora water extract on model rats. The experimental protocol strictly adhered to the guidelines of the Ethics Committee of Animal Research of Guangdong Pharmaceutical University (Approval: gdpulac2022132). Quality markers with anti-inflammatory activities were recognized by network pharmacology analysis. Subsequently, a reliable method for simultaneously quantifying six components was established. As a result, the pathological injury on lung and colon tissues of model rats induced by LPS were improved by C. nudiflora water extract. Compared with model group, C. nudiflora extract had decreased the release of proinflammatory factors in serum, including TNF-α and IL-6, and reduced the NO (P < 0.01). Network pharmacological analysis obtained 83 chemical components, 466 component targets, 584 disease targets and 129 action targets, which were mainly concentrated in 624 biological processes such as inflammatory response and positive regulation of nitric oxide biosynthesis, as well as 162 pathways such as phosphatidylinositol-3-hydroxykinase-protein kinase (PI3K-Akt) signal pathway and Janus kinase-signal transducer and activator of transcription 3 (JAK-STAT3) signal pathway. Upon analyzing their specificity, effectiveness, detectability, and quantity transfer rule from herb to extract, 6-hydroxyluteolin 7-glucoside, cynaroside, caffeic acid, acteoside, isoacteoside, and forsythoaside B were identified as the quality markers. Their anti-inflammatory activities were verified with LPS-induced inflammation model of RAW264.7 cells. Then contents of these 6 quality markers in 16 batches of C. nudiflora were determined. Thereby, the anti-inflammatory efficacy of C. nudiflora extract were confirmed in vivo, 6-hydroxyluteolin 7-glucoside, cynaroside, caffeic acid, acteoside, isoacteoside, and forsythoaside B were identified as anti-inflammatory ingredients, which can be used as quality control indicators for the herb and related formulation.

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.

The effect of porcine SIRT5 on replication of foot and mouth disease virus type O(FMDV-O)and the underlying regulatory mechanism were investigated.Western blot and RT-qPCR analyses were employed to monitor expression of endoge-nous SIRT5 in PK-15 cells infected with FMDV-O.Three pairs of SIRT5-specific siRNAs were synthesized.Changes to SIRT5 and FMDV-O protein and transcript levels,in addition to virus copy numbers,were measured by western blot and RT-qPCR analyses.PK-15 cells were transfected with a eukaryotic SIRT5 expression plasmid.Western blot and RT-qPCR analyses were used to explore the impact of SIRT5 overexpression on FMDV-O replication.Meanwhile,RT-qPCR analysis was used to detect the effect of SIRT5 overexpression on the mRNA expression levels of type I interferon-stimulated genes induced by SeV and FMDV-O.The results showed that expression of SIRT5 was up-regulated in PK-15 cells infected with FMDV-O and siRNA interfered with SIRT5 to inhibit FMDV-O replication.SIRT5 overexpression promoted FMDV-O replication.SIRT5 over-expression decreased mRNA expression levels of interferon-stimulated genes induced by SeV and FMDV-O.These results suggest that FMDV-O infection stimulated expression of SIRT5 in PK-15 cells,while SIRT5 promoted FMDV-O rep-lication by inhibiting production of type I interferon-stimula-ted genes.These findings provide a reference to further ex-plore the mechanism underlying the ability of porcine SIRT5 to promote FMDV-O replication.

Objective:To explore potential predictors of refractory Mycoplasma pneumoniae pneumonia (RMPP) in early stage. Methods:The prospective multicenter study was conducted in Zhejiang, China from May 1 st, 2019 to January 31 st, 2020. A total of 1 428 patients with fever >48 hours to <120 hours were studied. Their clinical data and oral pharyngeal swab samples were collected; Mycoplasma pneumoniae DNA in pharyngeal swab specimens was detected. Patients with positive Mycoplasma pneumoniae DNA results underwent a series of tests, including chest X-ray, complete blood count, C-reactive protein, lactate dehydrogenase (LDH), and procalcitonin. According to the occurrence of RMPP, the patients were divided into two groups, RMPP group and general Mycoplasma pneumoniae pneumonia (GMPP) group. Measurement data between the 2 groups were compared using Mann-Whitney U test. Logistic regression analyses were used to examine the associations between clinical data and RMPP. Receiver operating characteristic (ROC) curves were used to analyse the power of the markers for predicting RMPP. Results:A total of 1 428 patients finished the study, with 801 boys and 627 girls, aged 4.3 (2.7, 6.3) years. Mycoplasma pneumoniae DNA was positive in 534 cases (37.4%), of whom 446 cases (83.5%) were diagnosed with Mycoplasma pneumoniae pneumonia, including 251 boys and 195 girls, aged 5.2 (3.3, 6.9) years. Macrolides-resistant variation was positive in 410 cases (91.9%). Fifty-five cases were with RMPP, 391 cases with GMPP. The peak body temperature before the first visit and LDH levels in RMPP patients were higher than that in GMPP patients (39.6 (39.1, 40.0) vs. 39.2 (38.9, 39.7) ℃, 333 (279, 392) vs. 311 (259, 359) U/L, both P<0.05). Logistic regression showed the prediction probability π=exp (-29.7+0.667×Peak body temperature (℃)+0.004×LDH (U/L))/(1+exp (-29.7+0.667×Peak body temperature (℃)+0.004 × LDH (U/L))), the cut-off value to predict RMPP was 0.12, with a consensus of probability forecast of 0.89, sensitivity of 0.89, and specificity of 0.67; and the area under ROC curve was 0.682 (95% CI 0.593-0.771, P<0.01). Conclusion:In MPP patients with fever over 48 to <120 hours, a prediction probability π of RMPP can be calculated based on the peak body temperature and LDH level before the first visit, which can facilitate early identification of RMPP.

Electrochemiluminescence(ECL)refers to the process of luminescence triggered by high-energy electron transfer between intermediate products during the oxidation-reduction reaction on the electrode surface.In the co-reactants involved ECL process,the presence of co-reaction accelerators can effectively catalyze the decomposition of co-reactants,leading to the generation of abundant free radical intermediates,thereby significantly enhancing the ECL signals.This plays a critical role in constructing simple,sensitive,and efficient ECL sensing platforms.This review focused on the novel co-reaction accelerators developed in recent years.Based on different types of co-reaction accelerator materials,including single atom catalysts,metal-based nanomaterials,polymers and other materials,the ECL reaction process and signal enhancement mechanisms,as well as their relevant applications in constructing ECL sensing platforms,were elucidated.Furthermore,the current research challenges and development prospect of co-reaction accelerators were also discussed.