Objective In order to shorten the transparency time of clear,unobstructed brain imaging cocktails and computational analysis(CUBIC),improve the transparency efficiency,and explore the possibility of applying hydrophilic tissue transparency technique,this study was conducted to optimize the perfusion of CUBIC technique and compare it with four hydrophilic tissue clearing method in terms of tissue transparency effect,transparency time,area change,volume change and adeno-associated virus(AAV)fluorescence retention.Methods Brain,liver,spleen and kidney of 6 adult Institute of Cancer Research(ICR)mice were subjected to clearing treatment by SeeDB,FRUIT,ScaleS and CUBIC method,respectively.The area and gray value of the samples were measured by Image J 1.8.0,and the volume before and after transparency was measured by drainage method to compare the transparency effect,time and size deformation of each group.Perfusion optimization of the CUBIC was performed by improving the perfusion rate with the optimal perfusion dose,each group of the experimental sample size was 6.Fluorescence preservation by different techniques was evaluated by injecting AAV in the motor cortex of 16 adult mice and taking the cervical spinal segments for transparency treatment after four weeks,and the fluorescence photographs were measured by Image J 1.8.0 to measure the mean fluorescent intensity.Results The optimal perfusion rate and dose of CUBIC was 15 ml/min and 200 ml respectively.For transparency ability and speed,the perfusion CUBIC had the lowest mean gray value and took the shortest time,while CUBIC consumed the longest time,and SeeDB,FRUIT,and ScaleS did not show good transparency ability.In terms of area and volume changes,several techniques showed different degrees of expansion after transparency of tissues or organs.In terms of fluorescence retention,perfusion CUBIC showed the best retention of green fluorescent protein(GFP)fluorescence signal,followed by CUBIC,ScaleS,FRUIT,and SeeDB.Conclusion Perfusion CUBIC technique shows the best tissue transparency,the shortest transparency time,and the most AAV fluorescence retention compared with other techniques.

Diabetic retinopathy (DR) is a diabetic ocular complication that can lead to poor vision and blindness. This experiment aimed to investigate the ameliorative effect and its mechanism of Panax notoginseng saponins (PNS) eye drops on streptozotocin (STZ)-induced non-proliferative diabetic retinopathy (NPDR) in rats. All experiments were approved by the Animal Research Committee of Shanghai University of Traditional Chinese Medicine. Animal welfare and the animal experimental protocols were strictly consistent with related ethics regulations of Shanghai University of Traditional Chinese Medicine (No. PZSHUTCM 211115004). Diabetes mellitus was induced by a single intraperitoneal injection of STZ into rats. Two months later, PNS solution was dropped binocular twice per day at 6 h intervals at dose of 20, 40, and 80 mg·kg^-1 continuously for 1 month. The morphological structure and activation of microglia of the retina were observed by hematoxylin-eosin staining and immunohistochemical assay. The disruption of the blood-retina barrier (BRB) was conducted by the Evans blue dye leakage assay. The number of acellular capillaries in the retina was assessed by digesting and hematoxylin-eosin staining assay. The number of retinal leukocyte adhesion was observed by fluorescein isothiocyanate-coupled concanavalin A lectin labeling assay. The serum expression of inflammatory factors was measured using enzyme-linked immunosorbent assay. Western blot experiments were used to detect the expression of relevant proteins in retinal tissue and transcriptional activation of nuclear factor kappa-B (NF-κB). The results revealed that PNS eye drops significantly increased the thickness of the retina, decreased the number of acellular capillaries, and up-regulated the expression of the tight junction proteins claudin-1 and occludin, thereby alleviating BRB damage. In addition, PNS eye drops were also able to significantly reduce leukocyte adhesion and microglia activation, the expression of inflammatory factors in serum, and the nuclear translocation of retinal p65 proteins, effectively inhibiting the occurrence of retinal inflammation. The above results showed that PNS eye drops played a role in improving NPDR by inhibiting the activation of the NF-κB signaling pathway and reducing inflammation.

Objective To investigate the efficacy and safety of nusinersen sodium in the treatment of children with spinal muscular atrophy(SMA).Methods A retrospective analysis was conducted on the clinical data of 50 children with 5q SMA who received nusinersen sodium treatment and multidisciplinary treatment management in Shanxi Children's Hospital from February 2022 to February 2024.Results Compared with the baseline data,67%(8/12),74%(35/47),and 74%(35/47)of the SMA children had a clinically significant improvement in the scores of Philadelphia Infant Test of Neuromuscular Disorders,Hammersmith Functional Motor Scale Expanded,and Revised Upper Limb Module,respectively,and the distance of 6-minute walking test increased from 207.00(179.00,281.50)meters to 233.00(205.25,287.50)meters(P<0.05)after nusinersen sodium treatment.Of all 50 children with SMA,24(48%)showed good tolerability after administration,with no significant or persistent abnormalities observed in 2 034 laboratory test results,and furthermore,there were no serious or immunological adverse events related to the treatment.After treatment,there was a significant change in forced vital capacity as a percentage of the predicted value in 27 children with restrictive ventilatory dysfunction,as well as a significant change in the level of 25-(OH)vitamin D in 15 children with vitamin D deficiency(P<0.05).Conclusions For children with SMA,treatment with nusinersen sodium can continuously improve the response rates of motor function scales,with good tolerability and safety.

Cold agglutinins(CA),autoantibodies against the antigen I or i on the surface of red blood cells,are mainly of IgM class,and the majority have κ light chains.They can lead to red blood cell agglutination at decreased body temperature and are usually associated with infections,drug reactions,autoimmune diseases,and hematological malignancies.However,solid tumors with CA are rare.We reported two cases of CA in the peripheral blood of patients with solid tumors.Peripheral complete blood cell count of the patients at admission showed reduced erythrocyte count and hematocrit,mismatching between erythrocyte count and hemoglobin,abnormally elevated levels of mean corpuscular hemoglobin and mean cell hemoglobin concentration.Peripheral blood smear showed erythrocyte aggregation.After the sample was preheated at 37 ℃ for 30 min,the reversibility of red blood cell aggregation was observed,and the erythrocyte parameters were corrected.
Humans ; Autoantibodies/isolation & purification* ; Female ; Breast Neoplasms/immunology* ; Ovarian Neoplasms/immunology*

ObjectiveTo explore the influencing factors of different scores on predicting death risk of extremely low birth weight infants （ELBWI）. MethodsA total of 186 cases of ELBWI admitted by the Children's Hospital affiliated to Nanjing Medical University and the Lishui Branch of the Affiliated Zhongda Hospital of Southeast University were admitted from January 1， 2019 to January 1， 2021， and 125 ELBWIs were finally included after screening by inclusion and exclusion criteria. There were 47 cases in the death group and 78 cases in the survival group. General data and the items of score for neonatal acute physiology version Ⅱ （SNAP-Ⅱ）， simplified version of the score for neonatal acute physiology perinatal extension （SNAPPE-Ⅱ）， clinical risk index for babies （CRIB）， clinical risk index for babies Ⅱ （CRIB-Ⅱ） and the national critical illness score （NCIS） were collected. Univariate and multivariate analysis was performed and nomogram was evaluated using receiver operating characteristic curve （ROC）. ResultsIt was found that systolic blood pressure， maximum inhaled oxygen concentration， BE value and birth weight were important factors in ELBWI mortality risk assessment ［systolic blood pressure OR： 0.968， 95%CI： 0.938-0.999， P=0.043； maximum inhaled oxygen concentration OR： 1.020， 95%CI： 1.006-1.034， P=0.006； BE OR： 0.868， 95%CI： 0.786-0.959， P=0.005； birth weight OR： 0.994， 95%CI： 0.991-0.997， P=0.000］. ROC showed that the area under the curve of the above four variables is 0.71， and the 95% confidence interval is 0.610-0.799， which is better than CRIB score. ConclusionLower systolic blood pressure， higher inhaled oxygen concentration， higher BE and lower birthweight are important influencing factors to predict the death risk of ELBWI. The above four items should be included in the newly developed score assessment to obtain a more effective ELBWI prediction system.

Pregnancy ; Female ; Humans ; Milk, Human ; Overweight/genetics* ; Fatty Acids, Unsaturated ; Fatty Acids ; Adiponectin/genetics*

OBJECTIVE: To observe the clinical effect of acupuncture at sphenopalatine ganglion combined with conventional acupuncture for episodic cluster headache (CH). METHODS: One hundred and eighty patients with episodic CH were randomly divided into a combined group (60 cases, 3 cases dropped off)，an acupuncture group (60 cases, 2 cases dropped off) and a sphenopalatine ganglion group (60 cases, 2 cases dropped off and 1 case was removed). The patients in the acupuncture group were treated with conventional acupuncture at Touwei (ST 8), Yintang (GV 24⁺), Yangbai (GB 14), Hegu (LI 4), etc., once a day, 6 times a week. The patients in the sphenopalatine ganglion group were treated with acupuncture at sphenopalatine ganglion, once every other day, 3 times a week. On the basis of the conventional acupuncture, the combined group was treated with acupuncture at sphenopalatine ganglion once every other day. Two weeks were taken as a course of treatment, and 3 courses of treatment were required in the 3 groups. The score of visual analogue scale (VAS), the number of headache attacks per week, the duration of each headache attack and the score of migraine-specific quality of life questionnaire version 2.1 (MSQ) were observed before and after treatment and in follow-up of 3 months after treatment. The clinical efficacy of each group was compared. RESULTS: After treatment and in follow-up, the VAS score of headache, the number of headache attacks per week, the duration of each headache attack, and each various scores and the total score of MSQ of each group were lower than those before treatment (P<0.01). Except that the number of headache attacks per week in the combined group was lower than the sphenopalatine ganglion group (P<0.01), other indexes in the combined group were lower than the other two groups (P<0.05, P<0.01). The total effective rate in the combined group was 93.0% (53/57), which was higher than 75.9% (44/58) in the acupuncture group and 73.7% (42/57) in the sphenopalatine ganglion group（P<0.05, P<0.01). CONCLUSION Acupuncture at sphenopalatine ganglion combined with conventional acupuncture could reduce the degree of pain in patients with episodic CH, reduce the number and duration of headache attacks, and improve the quality of life of patients. It is more effective than simple conventional acupuncture or acupuncture at sphenopalatine ganglion alone.
Acupuncture Points ; Acupuncture Therapy ; Cluster Headache/therapy* ; Headache/therapy* ; Humans ; Quality of Life ; Treatment Outcome

ObjectiveTo explore effect of Huanglian Jiedutang （HLJDT） on autophagy-related protein expression in septic mice with liver injury induced by cecal ligation and puncture （CLP）. MethodSixty eight-week-old C57BL/6 mice were randomly divided into four groups， namely， the sham operation group， model group， and low- （1.44 g∙kg^-1） and high-dose （2.88 g∙kg^-1） HLJDT groups， with 15 in each group. The septic model was established by CLP after the last administration of HLJDT for three successive days. The survival rate of mice with 24 h was observed. The mice were sacrificed 12 h after operation for collecting the serum and liver tissue. The levels of serum interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， and interleukin-1β （IL-1β） were detected by enzyme-linked immunosorbent assay （ELISA）， and the levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum by biochemical method. The pathological changes in liver tissue were observed by hematoxylin-eosin （HE） staining， and the apoptosis index （AI） of hepatocytes by TdT-mediated dUTP-biotin nick end-labeling （TUNEL）. The expression levels of protein high-mobility group box 1 protein （HMGB1）， Beclin1， and microtubule-associated protein 1 light chain 3 （LC3）-Ⅱ/Ⅰ in the liver tissue were assayed by Western blot. ResultCompared with the sham operation group， the model group showed reduced survival rate at 12 and 24 h， elevated IL-6， TNF-α， and IL-1β levels， enhanced AST and ALT activities （P<0.05）， hepatocyte swelling， inflammatory cell infiltration， and apoptosis， and up-regulated HMGB1 （P<0.05）， Beclin1， and LC3-Ⅱ/Ⅰ （P<0.05）. Compared with the model group， each medication group exhibited increased survival rate at 12 and 24 h， lowered IL-6 and TNF-α levels， weakened AST and ALT activities （P<0.05）， alleviated liver injury and apoptosis （P<0.05）， down-regulated HMGB1 expression （ P<0.05）， and up-regulated Beclin1 and LC3-Ⅱ/Ⅰ （P<0.05）. ConclusionHLJDT alleviates the liver injury of septic mice possibly by inducing autophagy and inhibiting apoptosis.

Based on network pharmacology and in vivo experiment, this study explored the therapeutic effect of Tetrastigma hemsle-yanum(SYQ) on sepsis and the underlying mechanism. The common targets of SYQ and sepsis were screened out by network pharmacology, and the "SYQ-component-target-sepsis" network was constructed. The protein-protein interaction(PPI) network was established by STRING. Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment were performed based on DAVID to predict the anti-sepsis mechanism of SYQ. The prediction results of network pharmacology were verified by animal experiment. The network pharmacology results showed that the key anti-sepsis targets of SYQ were tumor necrosis factor(TNF), interleukin(IL)-6, IL-1β, IL-10, and cysteinyl asparate specific proteinase 3(caspase-3), which were mainly involved in Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor kappaB(NF-κB) signaling pathway. The results of animal experiment showed that SYQ can decrease the content of C-reactive protein(CRP), procalcitonin(PCT), lactate dehydrogenase(LDH), IL-6, TNF-α, and IL-1β, increase the content of IL-10, and down-regulate the protein levels of Bcl-2-associa-ted X(Bax)/B-cell lymphoma 2(Bcl2), cleaved caspase-3, TLR4, MyD88, and p-NF-κB p65/NF-κB p65. In summary, SYQ plays an anti-inflammatory role in the treatment of sepsis by acting on the key genes related to inflammation and apoptosis, such as TNF-α, IL-6, IL-lβ, IL-10, Bax, Bcl2, and cleaved caspase-3. The mechanism is the likelihood that it suppresses the TLR4/MyD88/NF-κB signaling pathway, which verifies relative prediction results of network pharmacology.
Animals ; Anti-Inflammatory Agents/therapeutic use* ; C-Reactive Protein ; Caspase 3/metabolism* ; Interleukin-10 ; Interleukin-6/metabolism* ; Lactate Dehydrogenases/metabolism* ; Myeloblastin/metabolism* ; Myeloid Differentiation Factor 88/metabolism* ; NF-kappa B/metabolism* ; Network Pharmacology ; Procalcitonin/therapeutic use* ; Sepsis/genetics* ; Toll-Like Receptor 4/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; bcl-2-Associated X Protein/metabolism*

Objective: To analyze the association between different types of obesity, BMI, and waist circumference (WC) and the risk of type 2 diabetes mellitus (T2DM) in Chinese adults. Methods: Based on the China Health and Nutrition Survey data in 2018, different types of obesity were defined across different BMI and WC combinations based on Criteria of weight for adults (WS/T 428-2013), including non-obesity, simple general obesity, simple central obesity, and complex obesity. The associations of different types of obesity, BMI, and WC with T2DM risk were explored using two-level mixed-effects logistic regression and restricted cubic spline models. Results: A total of 7 030 subjects aged 18-65 were included in this study. The prevalence of general obesity and central obesity were 16.29% (502/3 082), 42.28% (1 303/3 082) in males, and 14.41% (569/3 948), 37.87% (1 495/3 948) in females. The prevalence of complex obesity was 14.70% and 12.97% in males and females, respectively. The prevalence of T2DM was 11.28% in participants, and the prevalence in complex obesity (18.98%, 22.07%) was higher than in simple central obesity (16.24%, 15.26%) and non-obesity (9.65%, 5.18%) in males and females, respectively. Multilevel regressions showed that males with simple central obesity and complex obesity had 1.61 (95%CI: 1.24-2.08) and 2.11 (95%CI: 1.56-2.86) times the odds of T2DM, respectively, as compared with the non-obesity; and the odds of T2DM in females were 2.70 (95%CI: 1.16-6.28) times for simple general obesity, 2.62 (95%CI: 2.01-3.40) times for simple central obesity, and 4.47 (95%CI: 3.35-5.98) times for complex obesity. A nearly linear positive association was observed between BMI and T2DM risk. WC was also positively associated with T2DM risk and a non-linearly increased risk in females (P for non-linear=0.024). The risk of T2DM increased when BMI ≥22.5 kg/m² and 23.0 kg/m², WC ≥85.0 cm and 80.0 cm in males and females, respectively. Conclusions: Complex obesity adults are more likely to suffer from T2DM. The risk of T2DM increases significantly when BMI is at the normal high values and waist circumference at the stage of pre-central obesity.
Adult ; Female ; Male ; Humans ; Diabetes Mellitus, Type 2/epidemiology* ; Prevalence ; Obesity, Abdominal/epidemiology* ; Obesity/epidemiology* ; China/epidemiology*