Objective:To investigate neuroprotective effect of alpinetin on ischemic stroke(IS)rats by regulating Shh/Gli1 signaling pathway.Methods:Fifteen rats were randomly collected as control group,remaining rats were used to construct an IS model.Rats that successfully modeling were randomly grouped into model group,alpinetin group(5 mg/kg),Shh/Gli1 signaling pathway inhibitor cycloparamide group(15 mg/kg)and alpinetin+cycloparamide group(5 mg/kg alpinetin+15 mg/kg cycloparamide),with 15 rats in each group,and administered once a day for two consecutive weeks,control group and model group were given equal amounts of physiological saline.Zea-Longa scoring method was applied to evaluate neural function;ELISA was applied to detect inflammatory factors levels;mass of brain tissue was weighed and water content of brain tissue was calculated;TTC staining was applied to measure volume of cerebral infarction;HE staining was applied to observe nerve cell damage;TUNEL staining was applied to detect neuronal apoptosis;qRT-PCR and Western blot were used to detect mRNA and protein expressions of Shh and Gli1.Results:There were no abnormalities in hippocampal tissue of control group,while hippocampal tissue structure of model group rats was abnor-mal,with disordered cell arrangement and nuclear pyknosis of nerve cells,cell damage rate,Zea-Longa score,infarct volume,IL-1β,IL-6,TNF-α,brain tissue water content,and cell apoptosis rate in model group were obviously higher than control group(P<0.05),mRNA and protein levels of Shh and Gli1 were obviously decreased(P<0.05);compared with model group,cells in alpinetin group were arranged more neatly,and phenomenon of neuronal cell nucleus pyknosis was improved,cell damage rate,Zea-Longa score,infarct volume,IL-1β,IL-6,TNF-α,brain tissue water content,and cell apoptosis rate were obviously decreased(P<0.05),mRNA and protein levels of Shh and Gli1 were obviously increased(P<0.05),trend of above indicators in cyclophosphamide group was opposite,ciclopramide reversed neuroprotective effect of alpinetin on IS rats.Conclusion:Alpinetin may exert neuroprotective effects on IS rats by activating Shh/Gli1 signaling pathway.

Since the approval of gemtuzumab ozogamicin,an antibody-drug conjugate(ADC)targeting CD33 in 2000,13 ADC drugs have been approved by the FDA.Although these drugs have clearly improved the survival of patients with various types of advanced cancers,their significant toxicity has compromised their therapeutic benefits.The adverse reactions of ADC drugs are complex and include on-target and off-target toxicities,where the payload drug is a determining factor.Antibody and linker may also affect the degree of toxicity.Combination therapy becomes an important strategy in anticancer treatment because of its increased efficiency,but treatment-related adverse reactions also increase accordingly.This review comprehensively analyzes the toxicity mechanisms of current ADC drugs and proposes various optimization strategies,including but not limited to optimizing linker molecules,upgrading antibody design,and changing drug administration strategies,to improve the overall safety profile of ADC drugs.

Objective:To evaluate the diagnostic efficacy and practicality of the Jaundice color card (JCard) as a screening tool for neonatal jaundice.Methods:Following the standards for reporting of diagnostic accuracy studies (STARD) statement, a multicenter prospective study was conducted in 9 hospitals in China from October 2019 to September 2021. A total of 845 newborns who were admitted to the hospital or outpatient department for liver function testing due to their own diseases. The inclusion criteria were a gestational age of ≥35 weeks, a birth weight of ≥2 000 g, and an age of ≤28 days. The neonate′s parents used the JCard to measure jaundice at the neonate′s cheek. Within 2 hours of the JCard measurement, transcutaneous bilirubin (TcB) was measured with a JH20-1B device and total serum bilirubin (TSB) was detected. The Pearson′s correlation analysis, Bland-Altman plots and the receiver operating characteristic (ROC) curve were used for statistic analysis.Results:Out of the 854 newborns, 445 were male and 409 were female; 46 were born at 35-36 weeks of gestational age and 808 were born at ≥37 weeks of gestational age. Additionally, 432 cases were aged 0-3 days, 236 cases were aged 4-7 days, and 186 cases were aged 8-28 days. The TSB level was (227.4±89.6) μmol/L, with a range of 23.7-717.0 μmol/L. The JCard level was (221.4±77.0) μmol/L and the TcB level was (252.5±76.0) μmol/L. Both the JCard and TcB values showed good correlation ( r=0.77 and 0.80, respectively) and agreements (96.0% (820/854) and 95.2% (813/854) of samples fell within the 95% limits of agreement, respectively) with TSB. The JCard value of 12 had a sensitivity of 0.93 and specificity of 0.75 for identifying a TSB ≥205.2?μmol/L, and a sensitivity of 1.00 and specificity of 0.35 for identifying a TSB ≥342.0?μmol/L. The TcB value of 205.2?μmol/L had a sensitivity of 0.97 and specificity of 0.60 for identifying TSB levels of 205.2 μmol/L, and a sensitivity of 1.00 and specificity of 0.26 for identifying TSB levels of 342.0 μmol/L. The areas under the ROC curve (AUC) of JCard for identifying TSB levels of 153.9, 205.2, 256.5, and 342.0 μmol/L were 0.96, 0.92, 0.83, and 0.83, respectively. The AUC of TcB were 0.94, 0.91, 0.86, and 0.87, respectively. There were both no significant differences between the AUC of JCard and TcB in identifying TSB levels of 153.9 and 205.2 μmol/L (both P>0.05). However, the AUC of JCard were both lower than those of TcB in identifying TSB levels of 256.5 and 342.0 μmol/L (both P<0.05). Conclusions:JCard can be used to classify different levels of bilirubin, but its diagnostic efficacy decreases with increasing bilirubin levels. When TSB level are ≤205.2 μmol/L, its diagnostic efficacy is equivalent to that of the JH20-1B. To prevent the misdiagnosis of severe jaundice, it is recommended that parents use a low JCard score, such as 12, to identify severe hyperbilirubinemia (TSB ≥342.0 μmol/L).

Objective:To estimate the influence of metabolic parameters in 18F-FDG PET/CT and clinically relevant indicators on the prognosis of patients with cervical cancer. Methods:A total of 174 patients with cervical cancer (age (53.6±11.1) years) who underwent baseline 18F-FDG PET/CT examination in the Affiliated Hospital of Qingdao University from May 2011 to December 2020 were retrospectively collected. Metabolic parameters (metabolic tumor volume of primary lesion (MTV p), total lesion glycolysis of primary lesion (TLG p), MTV sum of total lesions (MTV total) in the whole body, TLG sum of total lesions (TLG total)) and clinical parameters (International Federation of Gynecology and Obstetrics (FIGO) stage, tumor maximum diameter ( Dmax), et al) were collected. Cox regression and Kaplan-Meier method were performed to evaluate the prognostic and predictive values of those parameters. Results:The follow-up time was 6-120 months, during which 52 patients (29.9%, 52/174) developed progression. The 5-year overall survival (OS), progression-free survival (PFS), local control (LC) and distant metastasis-free survival (DMFS) rates were 83.3%(145/174), 70.1%(122/174), 75.3%(131/174) and 82.8%(144/174), respectively. Cox regression showed that FIGO stage and MTV total were independent factors for predicting PFS, OS and LC (hazard ratio ( HR): 1.005-11.605, all P<0.05). FIGO stage and TLG total were independent factors for predicting DMFS ( HR: 1.002-12.258, all P<0.05). Conclusion:MTV total and FIGO stage are effective predictors of patients with cervical squamous cell carcinoma.

The syndrome of dampness stagnancy due to spleen deficiency (DSSD) is relatively common globally. Although the pathogenesis of DSSD remains unclear, evidence has suggested that the gut microbiota might play a significant role. Radix Astragali, used as both medicine and food, exerts the effects of tonifying spleen and qi. Astragalus polysaccharide (APS) comprises a macromolecule substance extracted from the dried root of Radix Astragali, which has many pharmacological functions. However, whether APS mitigates the immune disorders underlying the DSSD syndrome via regulating gut microbiota and the relevant mechanism remains unknown. Here, we used DSSD rats induced by high-fat and low-protein (HFLP) diet plus exhaustive swimming, and found that APS of moderate molecular weight increased the body weight gain and immune organ indexes, decreased the levels of interleukin-1β (IL-1β), IL-6, and endotoxin, and suppressed the Toll-like receptor 4/nuclear factor-‍κB (TLR4/NF-‍κB) pathway. Moreover, a total of 27 critical genera were significantly enriched according to the linear discriminant analysis effect size (LEfSe). APS increased the diversity of the gut microbiota and changed its composition, such as reducing the relative abundance of Pseudoflavonifractor and Paraprevotella, and increasing that of Parasutterella, Parabacteroides, Clostridium XIVb, Oscillibacter, Butyricicoccus, and Dorea. APS also elevated the contents of short-chain fatty acids (SCFAs). Furthermore, the correlation analysis indicated that 12 critical bacteria were related to the body weight gain and immune organ indexes. In general, our study demonstrated that APS ameliorated the immune disorders in DSSD rats via modulating their gut microbiota, especially for some bacteria involving immune and inflammatory response and SCFA production, as well as the TLR4/NF-κB pathway. This study provides an insight into the function of APS as a unique potential prebiotic through exerting systemic activities in treating DSSD.
Rats ; Animals ; NF-kappa B/metabolism* ; Spleen ; Gastrointestinal Microbiome ; Toll-Like Receptor 4 ; Polysaccharides/pharmacology* ; Astragalus Plant/metabolism* ; Immune System Diseases/drug therapy* ; Body Weight

Detailed characterizations of genomic alterations have not identified subtype-specific vulnerabilities in adult gliomas. Mapping gliomas into developmental programs may uncover new vulnerabilities that are not strictly related to genomic alterations. After identifying conserved gene modules co-expressed with EGFR or PDGFRA (EM or PM), we recently proposed an EM/PM classification scheme for adult gliomas in a histological subtype- and grade-independent manner. By using cohorts of bulk samples, paired primary and recurrent samples, multi-region samples from the same glioma, single-cell RNA-seq samples, and clinical samples, we here demonstrate the temporal and spatial stability of the EM and PM subtypes. The EM and PM subtypes, which progress in a subtype-specific mode, are robustly maintained in paired longitudinal samples. Elevated activities of cell proliferation, genomic instability and microenvironment, rather than subtype switching, mark recurrent gliomas. Within individual gliomas, the EM/PM subtype was preserved across regions and single cells. Malignant cells in the EM and PM gliomas were correlated to neural stem cell and oligodendrocyte progenitor cell compartment, respectively. Thus, while genetic makeup may change during progression and/or within different tumor areas, adult gliomas evolve within a neurodevelopmental framework of the EM and PM molecular subtypes. The dysregulated developmental pathways embedded in these molecular subtypes may contain subtype-specific vulnerabilities.
Humans ; Brain Neoplasms/pathology* ; Neoplasm Recurrence, Local/metabolism* ; Glioma/pathology* ; Neural Stem Cells/pathology* ; Oligodendrocyte Precursor Cells/pathology* ; Tumor Microenvironment

The biological samples of oral genetic diseases and rare diseases are extremely precious. Collecting and preserving these biological samples are helpful to elucidate the mechanisms and improve the level of diagnose and treatment of oral genetic diseases and rare diseases. The standardized construction of biobanks for oral genetic diseases and rare diseases is important for achieving these goals. At present, there is very little information on the construction of these biobanks, and the standards or suggestions for the classification and coding of biological samples from oral and maxillofacial sources, and this is not conducive to the standardization and information construction of biobanks for special oral diseases. This consensus summarizes the background, necessity, principles, and key points of constructing the biobank for oral genetic diseases and rare diseases. On the base of the group standard "Classification and Coding for Human Biomaterial" (GB/T 39768-2021) issued by the National Technical Committee for Standardization of Biological Samples, we suggest 76 new coding numbers for different of biological samples from oral and maxillofacial sources. We hope the consensus may promote the standardization, and smartization on the biobank construction as well as the overall research level of oral genetic diseases and rare diseases in China.

Objective:To investigate the value of 18F-FDG PET/CT metabolic parameters in predicting perineural invasion (PNI) in patients with non-metastatic rectal cancer. Methods:From August 2012 to April 2020, 81 patients (51 males, 30 females, median age: 63 years) who received PET/CT examination and pathologically confirmed as rectal cancer in the Affiliated Hospital of Qingdao University were retrospectively analyzed. The 18F-FDG PET/CT metabolic parameters including SUV max, metabolic tumor volume (MTV) and total lesion glycolysis (TLG), and clinicopathological factors including gender, age, carcinoembryonic antigen (CEA), carbohydrate antigen (CA)19-9, maximum tumor diameter, degree of differentiation, T stage, lymph node metastasis, and TNM stage were recorded. Mann-Whitney U test and χ2 test were used to compare the differences of each parameter between PNI positive group and PNI negative group. Multivariate logistic regression was used to analyze the independent predictor of positive PNI. ROC curve was used to analyze its predictive efficacy. Results:Of 81 patients, 32(39.51%) were PNI positive and 49(60.49%) were PNI negative. There were significant differences of T stage ( χ2=10.73, P=0.010), lymph node metastasis ( χ2=6.21, P=0.013), TNM stage ( χ2=7.61, P=0.022), MTV (14.6(10.4, 24.7)and 9.0(5.4, 14.5) cm 3; U=-3.48, P=0.001) and TLG (108.588(72.749, 182.707) and 65.365(35.593, 117.682) g; U=-2.79, P=0.005) between PNI positive group and PNI negative group. Multivariate logistic regression analysis showed that MTV was the independent predictor of positive PNI in non-metastatic rectal cancer patients (odds ratio ( OR)=1.130, 95% CI: 1.025-1.245, P=0.014). The optimal threshold of MTV was 9.53 cm 3 and AUC was 0.73 with the sensitivity of 81.82%(27/33) and the specificity of 59.18%(29/49). Conclusion:18F-FDG PET/CT metabolic parameter MTV can predict PNI in non-metastatic rectal cancer with high sensitivity.

Objective:To explore the effects of miRNA-373-3p (miR-373-3p) on the proliferation of nephroblastoma G401 cells through targeted regulation of CD44 expression.Methods:Bioinformatic method was used to predict the possible targeted genes of miR-373-3p based on bioinformatic databases including miRDB, miRanda, PITA and DIANA-microT. G401 cells were taken and transfected with miR-373-3p mimic, mimic negative control, miR-373-3p inhibitor or inhibitor negative control, respectively. Cell proliferation ability was detected by using CCK-8 assay. The number of clones was detected by using clone formation assay. The relative expression level of CD44 mRNA was detected by using real-time fluorescent quantitative polymerase chain reaction (qRT-PCR), and the expression level of CD44 protein was detected by using Western blotting. The dual luciferase gene reporter assay was carried out in HEK-293T cells to vertify the target gene of miR-373-3p.Results:Bioinformatic analysis indicated that CD44 was a targeted gene of miR-373-3p. After 24 h transfection, the proliferation activity of G401 cells in miR-373-3p mimic group was decreased compared with that in mimic negative control group (all P < 0.05). After 48 h transfection, the proliferation activity of tumor cells in miR-373-3p inhibitor group was increased compared with that inhibitor negative control group (all P < 0.05). The formed number of clones in miR-373-3p mimic group was reduced compared with that in the mimic negative control group (55.3±2.5 vs. 90.7±2.9), and the difference was statistically significant ( t = 14.57, P < 0.01). The formed number of clones in miR-373-3p inhibitor group was more than that in inhibitor negative control group (115.0±2.7 vs. 92.0±2.4), and the difference was statistically significant ( t = 8.86, P < 0.01). The dual-luciferase gene reporter assay showed that CD44 was a direct targeted gene of miR-373-3p. The relative expression levels of CD44 mRNA in miR-373-3P mimic and mimic negative control group were 0.62±0.03 and 1.00±0.01, respectively, and the difference was statistically significant ( t = 11.28, P < 0.01). The relative expression levels of CD44 mRNA in miR-373-3p inhibitor and inhibitor negative control group were 1.31±0.02 and 1.00±0.00, respectively, and the difference was statistically significant ( t = 12.65, P < 0.01). The CD44 protein expression was decreased in miR-373-3p mimic group, while increased in miR-373-3p inhibitor group. Conclusion:miR-373-3p can inhibit tumor cell proliferation by targeting CD44 in nephroblastoma.

Objective: To evaluate the efficacy and safety of different positions external physical vibration lithecbole (EPVL) therapy for ureteral calculi related renal colic. Methods: This study was a prospective multicenter randomized controlled trial. The inclusion criteria was that patients volunteered to participate in the trial and signed informed consent, patients’age ranged from 18-65 years old, ureteral calculi related with renal colic, stone diameter was less than 7 mm, patients were not treated with analgesia, antispasmodic drugs. The exclusion criteria was that combination of severe urinary tract infection, severe hydronephrosis, urinary malformation, severe hypertension, history of cerebrovascular disease, vital organ dysfunction, obesity (BMI>35 kg/m²), history of ureteral calculi exceeded 2 months, abnormal blood coagulation. Patients were randomized into observation group and control group using random number table method. The observation group and the control group were placed on the physical vibration stone arranging machine with head low foot high position and head high foot low position respectively. The inclination angle was 24°. The secondary vibrator vibrated for 6 minutes, then the patient took the prone position and opened the main, the secondary vibrator. The treatment is completed after 6 minutes of vibration. The analgesic effect, stone removal, follow-up effects and adverse reactions in the two groups was compared. We defined the pain relief rate as(VAS score before treatment-VAS score after treatment)/VAS score before treatment×100%. Results: A total of 100 patients were included in the study, 50 in the observation group and 50 in the control group. There were no statistical difference in the age of the two groups [(41.8±11.7)years and (46.6±13.9 years)], gender distribution [37(male)/13(female) and 42(male)/ 8(female)], location of stones (in the observation group, 19 cases in upper ureter, 7 cases in the middle ureter and 24 cases in the lower ureter; in the control group, 12 cases in the upper ureter, 3 cases in the middle ureter, and 35 in the lower ureter), left and right distribution of stones [21(right)/ 29 (left) and 22 (right)/ 28(left)], long diameter of stones [(5.2±0.9)mm and(5.1±1.1)mm], VAS scores before treatment (7.5±1.4 and 7.6±1.5), and readmission rate [22%(11/50)With 18%(9/50)], 1 week stone removal rate [70%(35/50) and 64%(32/50)]. The incidence of adverse reactions was 8%(4/50) in the observation group including 3 cases of nausea, 1 case of vomiting. The incidence of adverse reactions was 4% in the control group (2/50), which 2 cases showed nausea. The number of patients who chose EPVL, ESWL or surgery for the subsequent treatment in observation group was 35 cases, 9 cases, and 6 cases respectively. The number of patients who chose EPVL, ESWL or surgery for the subsequent treatment in the control group was 35 cases, 10 cases and 5 cases respectively. There was no significant difference between the two groups (P>0.05). The VAS score after treatment of the observation group was lower than that of the control group (2.4±1.3 and 3.7±1.5, P<0.01). The total effective rate of the observation group was higher than that of the control group [94%(47/50) and 76%(38/50), P<0.01]. 46 patients underwent urinary ultrasonography to check the degree of hydronephrosis. The reduction rate of hydronephrosis was higher in the observation group than in the control group [54.5%(18/33) and 30.8%(4/13)], but there was no significant difference between the two groups (P=0.146). In the upper ureteral calculi, the VAS score of the observation group was lower than that of the control group (2.4±0.3 and 3.9±0.4, P<0.01). There was no significant difference in the total effective rate between the two groups. In the lower ureteral calculi, the observation group had low VAS score after treatment. In the control group (2.4±0.2 and 3.5±0.2, P<0.01), there was no significant difference in the total effective rate between the two groups; the middle segment stones were less included (10 in total) and were not discussed. Conclusions The external physical vibration lithecbole adopts " head high and low position" has better effect than "head low foot high position" in treatment of ureteral calculi with renal colic, and security is acceptable.