ObjectiveTo investigate the therapeutic effect of sequential administration of Dihuang Baoyuan granules （DHBY， the prescription for consolidating body resistance） and Fuling Yunhua granules （FLYH， the prescription for treating symptoms） on spontaneous type 2 diabetes mellitus （T2DM） in mice. MethodsAccording to the fasting blood glucose （FBG） level， 12-week-old db/db mice were randomized into six groups： model， DHBY （18.02 g·kg^-1）， FLYH （14.80 g·kg^-1）， sequential administration 1 （SEQ-1， DHBY 18.02 g·kg^-1+FLYH 14.80 g·kg^-1）， sequential administration 2 （SEQ-2， FLYH 14.80 g·kg^-1+DHBY 18.02 g·kg^-1）， and dapagliflozin （Dapa， 1.3 mg·kg^-1）. The m/m mice in the same litter were selected as the normal group. The mice were administrated with corresponding drugs by gavage for 8 consecutive weeks. During the 8 weeks of drug administration and 2 weeks after withdrawal， the retinal thickness， FBG， hemoglobin A1c （HbA1c）， and insulin were determined， and histopathological changes of the pancreas， liver， kidney， and retina were observed by hematoxylin-eosin （HE） staining. ResultsCompared with the model group， SEQ-1 for 4 weeks lowered the FBG level （P<0.05）， raised the insulin level， decreased the triglyceride （TG） level （P<0.05）， increased the number of optic ganglion cells and diminished vacuolar degeneration of pancreatic islet and liver. SEQ-2 lowered FBG and HbA1c levels （P<0.05）， rose the insulin level， increased the retinal thickness and the number of optic ganglion cells （P<0.05）， and alleviated vacuolar degeneration of pancreatic islet and liver. Two weeks after drug withdrawal， Dapa tended to increase FBG and HbA1c compared with those at the time of drug withdrawal. However， the levels of FBG and HbA1c in the SEQ-2 group remained decreasing （P<0.05）. ConclusionSEQ-1 and SEQ-2 can lower the blood glucose level and ameliorate diabetic retinopathy， and SEQ-2 outperformed DHBY and FLYH in lowering the blood glucose level. Moreover， SEQ-2 can maintain the blood glucose-lowering effect after drug withdrawal.

ObjectiveTo investigate the therapeutic effect of sequential administration of Dihuang Baoyuan granules （DHBY， the prescription for consolidating body resistance） and Fuling Yunhua granules （FLYH， the prescription for treating symptoms） on spontaneous type 2 diabetes mellitus （T2DM） in mice. MethodsAccording to the fasting blood glucose （FBG） level， 12-week-old db/db mice were randomized into six groups： model， DHBY （18.02 g·kg^-1）， FLYH （14.80 g·kg^-1）， sequential administration 1 （SEQ-1， DHBY 18.02 g·kg^-1+FLYH 14.80 g·kg^-1）， sequential administration 2 （SEQ-2， FLYH 14.80 g·kg^-1+DHBY 18.02 g·kg^-1）， and dapagliflozin （Dapa， 1.3 mg·kg^-1）. The m/m mice in the same litter were selected as the normal group. The mice were administrated with corresponding drugs by gavage for 8 consecutive weeks. During the 8 weeks of drug administration and 2 weeks after withdrawal， the retinal thickness， FBG， hemoglobin A1c （HbA1c）， and insulin were determined， and histopathological changes of the pancreas， liver， kidney， and retina were observed by hematoxylin-eosin （HE） staining. ResultsCompared with the model group， SEQ-1 for 4 weeks lowered the FBG level （P<0.05）， raised the insulin level， decreased the triglyceride （TG） level （P<0.05）， increased the number of optic ganglion cells and diminished vacuolar degeneration of pancreatic islet and liver. SEQ-2 lowered FBG and HbA1c levels （P<0.05）， rose the insulin level， increased the retinal thickness and the number of optic ganglion cells （P<0.05）， and alleviated vacuolar degeneration of pancreatic islet and liver. Two weeks after drug withdrawal， Dapa tended to increase FBG and HbA1c compared with those at the time of drug withdrawal. However， the levels of FBG and HbA1c in the SEQ-2 group remained decreasing （P<0.05）. ConclusionSEQ-1 and SEQ-2 can lower the blood glucose level and ameliorate diabetic retinopathy， and SEQ-2 outperformed DHBY and FLYH in lowering the blood glucose level. Moreover， SEQ-2 can maintain the blood glucose-lowering effect after drug withdrawal.

Objective:To explore the characteristics and therapeutic strategies of Pott's puffy tumor（PPT）. Methods:The clinical data of two patients with PPT were retrospectively analyzed and combined with the literature, focusing on the comprehensive analysis of perioperative diagnosis and treatment strategies. Both patients underwent muti-disciplinary treatment, including timely administration of sufficient antibiotics capable of penetrating the blood-brain barrier. Early removal of PPT lesions was performed using a combined internal and external approach under nasal endoscopic guidance. Results:After standardized perioperative management, the symptoms of the two patients were completely relieved, with no recurrence after one=year follow=up. Postoperative complications such as frontal pain, numbness, local depression, or scar hyperplasiawere not present. Conclusion:PPT, being relatively rare and severe, requires careful attention. Key strategies for standardized perioperative management include multi-disciplinary consultation, timely and adequate antibiotic administration, and surgical intervention using a combined intranasal and extranasal endoscopic approach for lesion removal.
Humans ; Pott Puffy Tumor/complications* ; Retrospective Studies ; Tomography, X-Ray Computed ; Endoscopy/adverse effects* ; Postoperative Complications ; Anti-Bacterial Agents/therapeutic use* ; Frontal Sinusitis/complications*

OBJECTIVE To investigate the effect and mechanism of miR-152-3p on the resistance to paclitaxel(PTX)of PTX-resistant ovarian cancer cells(A2780T cells).METHODS ① Ovarian cancer parent cells(A2780 cells)and A2780T cells were treated with PTX(1.875,3.75,7.5,17 and 23 μmol·L-1)for 48 h.Cell viability was evaluated by MTT assay,and the 50%inhibitory concentration(IC50)and drug resistance index of A2780T cells were calculated.Western blotting was used to detect the expres-sions of resistance protein P-glycoprotein(P-gp),multidrug resistance related protein 1(MRP1)and adenosine triphosphate binding transporter G superfamily member 2(ABCG2).② Real-time fluorescent quantita-tive PCR(RT-qPCR)was used to detect the expressions of miR-152-3p in A2780 and A2780T cells.The lipid-mediated transient transfection technique was employed to transfect the miR-152-3p inhibitor to reduce miR-152-3p expression in A2780T cells(miR-152-3p inhibitor group),while the negative control(miR-152-3p NC)group was established.RT-qPCR was used to detect transfection efficiency,and the MTT method,scratch experiment,and flow cytometry were used to investigate the effects of the trans-fecting miR-152-3p inhibitor on survival,migration and apoptosis of A2780T cells.Western blotting was used to detect the protein expressions of Bax and Bcl-2 in A2780T cells.③ Bioinformatics analysis of databases including miRDB,Targetscan,miRWalk,and Starbase predicted the target genes of miR-152-3p that were verified by Western blotting to detect the protein expression of PTEN in A2780T cells of the miR-152-3p inhibitor and miR-152-3p NC groups,and RT-qPCR to detect the PTEN mRNA expression in A2780 and A2780T cells.Then,the lipid-mediated transient transfection technique was used to transfect PTEN siRNA to silence PTEN expression in A2780T cells(PTEN siRNA group).The siRNA negative control(siRNA NC)group was established.RT-qPCR was used to detect transfection efficiency,the MTT method was employed to measure the survival rate and IC50 value,and Western blotting was used to assess the protein expressions of P-gp,MRP1,and ABCG2 in A2780T cells after silencing PTEN expression.RESULTS ①After treatment with PTX,the cell survival rates were decreased in A2780 and A2780T cells(P<0.05),and the resistance index of A2780T cells was 2.8.Compared with A2780 cells,the protein expressions of P-gp and MRP1 and ABCG2 were highly expressed in A2780T cells(P<0.05,P<0.01).② RT-qPCR showed that the expression of miR-152-3p in A2780T cells was higher than that of A2780 cells(P<0.01).Compared with the miR-152-3p NC group,A2780T cell viability(P<0.05,P<0.01)and cell migration capability(P<0.05)were significantly inhibited,while the apoptosis rate increased(P<0.01)in miR-152-3p inhibitor group.Moreover,the protein expression of Bax was increased(P<0.01),but Bcl-2 decreased(P<0.05).③ Bioinformatics analysis suggested that PTEN was a target gene of the miR-152-3p,and the verified results showed that the PTEN protein expression in A2780T cells of the miR-152-3p inhibitor group was lower than that of the miR-152-3p NC group(P<0.05),and PTEN mRNA expression in A2780T cells was higher than that in A2780 cells(P<0.01).After silencing the expression of PTEN in A2780T cells,the cell viability was significantly reduced(P<0.05,P<0.01),while the IC50 value was reduced(P<0.01)compared with the siRNA NC group.In addition,the protein expressions of P-gp,MRP1 and ABCG2 were decreased(P<0.05,P<0.01).CONCLUSION miR-152-3p is highly expressed in A2780T cells,and down-regulation of its expression may inhibit proliferation and migration,prompt apoptosis and reduce the resistance to PTX of A2780T cells,which is made possible by inhibiting expression of its target gene PTEN.

Objective:To discuss the clinical characteristics,diagnosis,and treatment of Shwachman-Diamond syndrome(SDS),and to enhance the clinicians'awareness of the disease.Methods:The clinical materials of one patient diagnosed with SDS,primarily presented with neutropenia and elevated transaminase levels,confirmed by genetic testing were retrospectively analyzed.The clinical manifestations,genetic features,diagnosis,and treatment methods of SDS were analyzed complemented with the relevant literatures.Results:This patient was a male child,aged 27 months.His initial clinical presentations were neutropenia and elevated transaminase levels.The patient had previously experienced diarrhea when the patient was 3 months old,which improved after treated with oral pancreatic enzyme dispersion.Over the past six months,the patient had recurrent respiratory infections.Upon admission,the examination results showed there was dental enamel hypoplasia,and the imaging results showed the abnormal bone density in the long bones of the limbs.The genetic sequencing results showed a homozygous mutation in the Shwachman-Bodian-Diamond syndrome(SBDS)gene(c.258+2T>C).During hospitalization,the patient received the hepatoprotective care and granulocyte augmentation supportive treatment,leading to an improvement in his condition,and the patient was discharged.During a one-year follow-up,the patient's condition was stable.Conclusion:The typical presentation of the SDS patient includes diarrhea,liver function abnormalities,hematologic abnormalities,and skeletal anomalies,particularly neutropenia;there may also be developmental delays and involvement of the heart,liver,central nervous system,skeleton,and immune system.The genetic testing of suspected children is crucial,and it can aid in the early diagnosis and treatment of SDS patients.

To ensure proper dosage of a drug,analytical quantification of it in biofluid is necessary.Liquid chro-matography mass spectrometry(LC-MS)is the conventional method of choice as it permits accurate identification and quantification.However,it requires expensive instrumentation and is not appropriate for bedside use.Using soluble epoxide hydrolase(sEH)inhibitors(EC5026 and TPPU)as examples,we report development of a nanobody-based enzyme-linked immunosorbent assay(ELISA)for such small molecules and its use to accurately quantify the drug chemicals in human samples.Under optimized conditions,two nanobody-based ELISAs were successfully established for EC5026 and TPPU with low limits of detection of 0.085 ng/mL and 0.31 ng/mL,respectively,and two order of magnitude linear ranges with high precision and accuracy.The assay was designed to detect parent and two biologically active metabolites in the investigation of a new drug candidate EC5026.In addition,the ELISAs displayed excellent correlation with LC-MS analysis and evaluation of inhibitory potency.The results indicate that nanobody-based ELISA methods can efficiently analyze drug like compounds.These methods could be easily implemented by the bedside,in the field in remote areas or in veterinary practice.This work il-lustrates that nanobody based assays offer alternative and supplementary analytical tools to mass spectrometry for monitoring small molecule medicines during clinical development and therapy.At-tributes of nanobody based pharmaceutical assays are discussed.

OBJECTIVE: To compare the performance of Clear Cell Likelihood Score (ccLS) v1.0 and v2.0 in diagnosing clear cell renal cell carcinoma (ccRCC) from small renal masses (SRM). METHODS: We retrospectively analyzed the clinical data and MR images of patients with pathologically confirmed solid SRM from the First Medical Center of the Chinese PLA General Hospital between January 1, 2018, and December 31, 2021, and from Beijing Friendship Hospital of Capital Medical University and Peking University First Hospital between January 1, 2019 and May 17, 2021. Six abdominal radiologists were trained for use of the ccLS algorithm and scored independently using ccLS v1.0 and ccLS v2.0. Random- effects logistic regression modeling was used to generate plot receiver operating characteristic curves (ROC) to evaluate the diagnostic performance of ccLS v1.0 and ccLS v2.0 for ccRCC, and the area under curve (AUC) of these two scoring systems were compared using the DeLong's test. Weighted Kappa test was used to evaluate the interobserver agreement of the ccLS score, and differences in the weighted Kappa coefficients was compared using the Gwet consistency coefficient. RESULTS: In total, 691 patients (491 males, 200 females; mean age, 54 ± 12 years) with 700 renal masses were included in this study. The pooled accuracy, sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of ccLS v1.0 for diagnosing ccRCC were 77.1%, 76.8%, 77.7%, 90.2%, and 55.7%, as compared with 80.9%, 79.3%, 85.1%, 93.4%, 60.6% with ccLS v2.0, respectively. The AUC of ccLS v2.0 was significantly higher than that of ccLS v1.0 for diagnosis of ccRCC (0.897 vs 0.859; P < 0.01). The interobserver agreement did not differ significantly between ccLS v1.0 and ccLS v2.0 (0.56 vs 0.60; P > 0.05). CONCLUSION ccLS v2.0 has better performance for diagnosing ccRCC than ccLS v1.0 and can be considered for use to assist radiologists with their routine diagnostic tasks.
Female ; Male ; Humans ; Adult ; Middle Aged ; Aged ; Carcinoma, Renal Cell/diagnosis* ; Retrospective Studies ; Kidney ; Carcinoma ; Kidney Neoplasms/diagnosis*

Objective:To evaluate systematically the association between the c. 415>C polymorphism of NUDT15 gene and the toxicity of 6-mercaptopurine(6-MP)in children with acute lymphoblastic leukemia(ALL).Methods:The literatures in domestic and foreign databases were retrieved: PubMed, EmBase, Cochrane Library, CNKI, CBM, VIP Chinese Sci-tech Journal Database, and Wanfang Database.The language was limited to Chinese or English.A case-control study or cohort study of 6-MP treatment in pediatric ALL related to the toxicity of the NUDT15 gene c. 415>C polymorphism was included.The time of search was set from the establishment of the database to October 1st, 2020.Two researchers screened the literature independently, extracted data from the literature that met the inclusion criteria, and evaluated the quality of the included studies.The association between locus polymorphism and toxicity during 6-MP chemotherapy was analyzed by Meta analysis with Rev Man 5.3 and Stata 12.0 software.Results:Nine studies were finally included, eight of which were cohort studies and one was a case-control study, with a total of 1 068 patients.The results showed that under the five genetic models, the mutation at c. 415>C of NUDT15 gene was significantly associated with the risk of leukopenia and neutropenia( P<0.01), while hepatotoxicity was with no significant association between the occurrence risk of damage( P>0.05). Conclusion:The mutation at c. 415>C of NUDT15 gene significantly increased the incidence of leukopenia and neutropenia during 6-MP chemotherapy, while there was no significant effect on the occurrence of hepatotoxicity.

OBJECTIVE To establish a method for simultaneous determi nation of six components in Melastoma dodecandrum and investigate its correlation with antioxidant activity. METHODS Ultra high-performance liquid chromatography （UPLC）method was adopted. The contents of gallic acid ，protocatechuic acid ，isovitexin，rutin and ellagic acid in 23 batches of M. dodecandrum were determined by quantitative analysis of multi-components by single-marker （QAMS）method，using vitexin as the internal reference. Then 1，1-diphenyl-2-picrylhydrazyl（DPPH）free radical method ，2，2′-azino-bis（3-ethylbenzothiazoline-6-sulfonic acid ） diammonium salt （ABTS）method and ferric ion reduction/antioxidant power （FRAP）method were applied to determine the antioxidant activity of 23 batches of M. dodecandrum . Grey correlation analysis and bivariate correlation analysis were used to evaluate the correlation between six components and antioxidant activity. RESULTS The content of vitexin were 0.021％-0.182％. The contents of gallic acid ，protocatechuic acid ，isovitexin，rutin and ellagic acid by QAMS method were 0.008％-0.042％， 0.003％-0.023％，0.071％-0.283％，0.013％-0.140％ and 0.006％-0.021％，respectively. Compared with the results of external standard method ，P was greater than 0.05. Grey correlation analysis showed that the grey correlation coefficients between the contents of six components an d antioxidant activity was 0.727 6- 0.866 9. Bivariate correlation analysis showed that the contents of vitexin ，isovitexin and rutin were positively correlated with antioxidant activity （P＜0.05 or P＜0.01），the content of gallic acid and protocatechuic acid were negatively correlated with antioxidant activity （P＜0.05）. There was no significant correlation between ellagic acid and antioxidant activity.CONCLUSIONS QAMS method is successfully established for the simultaneous determination of six components in M. dodecandrum. The six components in M. dodecandrum are highly correlated with antioxidant activity.

Objective: To explore the value of CT texture analysis (CTTA) in differentiating the pathological grade of urothelial carcinoma of the bladder (UCB). Methods: A total of 53 lesions from 43 patients with bladder cancer confirmed by postoperative pathology were retrospectively analyzed, including 27 cases of high-grade urothelial carcinoma (HGUC) and 26 cases of low-grade urothelial carcinoma (LGUC). All the patients took pelvic CT and enhanced scanning in the same CT scanner with same scanning parameters. Lesions on both plain and enhanced CT images were delineated on software by two radiologists to extract the corresponding volumes of interest (VOI) and then 92 parameters based on feature classes were generated. The average values of two radiologists were obtained. The difference parameters between HGUC group and LGUC group were screened by nonparametric test, and the receiver operating characteristic (ROC) was drawn. The corresponding optimal thresholds were determined and diagnostic effect was assessed. Results: Nine difference texture parameters between HGUC group and LGUC group were selected, including 5 parameters on unenhanced images, namely, skewness, root mean squared, cluster shade, zone percentage and large area high gray level emphasis. There were 4 parameters on enhanced images, namely, skewness, kurtosis, cluster shade and zone percentage. The largest area under curve of 0.840±0.058 (95% CI 0.726-0.955) was obtained from skewness generated by VOI of unenhanced images. The cut-off value of skewness was 0.186 5, which permitted the diagnosis of HGUC with sensitivity of 92.59%, specificity of 73.08%, positive predictive value of 78.13%, negative predictive value of 90.48% and accuracy of 83.02%. Conclusion CTTA can effectively distinguish between LGUC and HGUC. Skewness from unenhanced CT images had the optimal diagnostic performance.