Application of 18F-FDG PET metabolic parameters in evaluating histopathologic grading of soft tissue sarcoma / 中华核医学与分子影像杂志

Objective:To evaluate the value of 18F-FDG PET metabolic parameters in predicting histopathological grade of soft tissue sarcoma (STS). Methods:From December 2012 to December 2021, 51 patients (26 males, 25 females, age range: 32-84 years) who underwent 18F-FDG PET/CT imaging before treatment and confirmed STS pathologically in the First Affiliated Hospital of Dalian Medical University were retrospectively collected. 18F-FDG PET metabolic parameters SUV max, metabolic tumor volume (MTV), total lesion glycolysis (TLG) and intertumoral FDG uptake heterogeneity (IFH) were measured. Kruskal-Wallis rank sum test was used to analyze the differences in metabolic parameters among different groups and Spearman rank correlation analysis was used to analyze the correlation of each metabolic parameter and histological grade. Logistic regression was used to screen and construct the prediction model for high-grade STS. ROC curve was plotted and Delong test was used to analyze the differences among AUCs. Results:The metabolic parameters SUV max, MTV, TLG and IFH were significantly different among French Federation of Cancer Centers Sarcoma Group (FNCLCC)Ⅰ( n=8), Ⅱ( n=10) and Ⅲ ( n=33) grade groups ( H values: 16.24, 10.52, 19.29 and 16.99, all P<0.05), and each metabolic parameter was positively correlated with histological grade ( rs values: 0.58, 0.45, 0.52, and 0.62, all P<0.05). Multivariate logistic regression analysis showed that SUV max(odds ratio ( OR)=1.27, 95% CI: 1.06-1.51, P=0.009) and IFH ( OR=6.83, 95% CI: 1.44-32.27, P=0.015) were independent risk indicators for high-grade STS. The prediction model constructed by combining SUV max and IFH had better diagnostic efficacy for differentiating high-grade STS with the AUC of 0.93, and the sensitivity of 93.9%(31/33) and the specificity of 16/18, respectively. The AUC of prediction model was significant different from SUV max, MTV, TLG and IFH (AUCs: 0.81, 0.78, 0.86 and 0.85; z values: 2.69, 2.53, 1.94 and 1.97, all P<0.05). Conclusions:The metabolic parameters SUV max, MTV, TLG and IFH are valuable predictors for histological grade of STS. The combination of SUV max and IFH may be a more meaningful method than using each of the above metabolic parameters alone.

Expression of alcohol dehydrogenase 1A and vascular endothelial growth factor-A in hepatocellular carcinoma / 安徽医科大学学报

Objective @#To investigate the expression , synergistic relationship and clinical significance of alcohol dehydrogenase (ADH1A) and vascular endothelial growth factor-A (VEGFA) in hepatocellular carcinoma (HCC) . @*Methods @#The expression and correlation of ADH1A and VEGFA in HCC and adjacent normal tissues were ana lyzed by GEPIA . TCGA and GSEA were used to analyze the pathway of ADH1A in HCC . The clinical and patho logical data of 84 patients with HCC were collected , and 54 patients with paracancer normal tissue samples were se lected as controls to analyze the correlation between ADH1A and VEGFA and clinicopathological parameters of HCC . Immunohistochemistry was used to detect the protein expression of ADH1A and VEGFA in cases and con trols , and the correlation between the expression of ADH1A and VEGFA and the clinical progression and prognosis of patients with HCC was analyzed based on clinical pathological parameters and Kaplan Meier.@*Results @#Bioinfor matics analysis found that ADH1A was low expressed in HCC and VEGFA was highly expressed in HCC , and there was a negative correlation between the two ( P < 0.001) ; immunohistochemical detection results showed that the expression of ADH1A in HCC tissue was lower than that in normal tissue adjacent to cancer (P < 0.01) while the expression rate of VEGFA in HCC tissue was significantly higher than that of normal tissue adjacent to cancer (P < 0.01) ; The recurrence rate of vascular thrombus and HCC patients in HCC group with high expression of ADH1A was lower (P < 0.05) . The proportion of tumor diameter > 5 cm , high TNM stage , microsatellite and G2 G3 dif ferentiation in HCC tissues in VEGFA high expression group was higher (P < 0.05) . Kaplan Meier survival analy sis showed that patients with high ADH1A expression and low VEGFA expression had a higher five year survival rate .@*Conclusion @#Low expression of ADH1A and high expression of VEGFA in tumor tissues of patients with HCC indicate tumor progression and can be used as one of the prognostic evaluation indicators for patients with HCC .

Targeting Ferroptosis to Enhance Radiosensitivity of Glioblastoma / 生物化学与生物物理进展

Glioblastoma (GBM), one of the most common malignant tumors in the central nervous system (CNS), is characterized by diffuse and invasive growth as well as resistance to various combination therapies. GBM is the most prevalent type with the highest degree of malignancy and the worst prognosis. While current clinical treatments include surgical resection, radiotherapy, temozolomide chemotherapy, novel molecular targeted therapy, and immunotherapy, the median survival time of GBM patients is only about one year. Radiotherapy is one of the important treatment modalities for GBM, which relies on ionizing radiation to eradicate tumor cells. Approximately 60% to 70% of patients need to receive radiotherapy as postoperative radiotherapy or neoadjuvant radiotherapy during the treatment process. However, during radiotherapy, the radioresistant effect caused by DNA repair activation and cell apoptosis inhibition impedes the therapeutic effect of malignant glioblastoma.Ferroptosis was first proposed by Dr. Brent R. Stockwell in 2012. It is an iron-dependent mode of cell death induced by excessive lipid peroxidation. Although the application of ferroptosis in tumor therapy is still in the exploratory stage, it provides a completely new idea for tumor therapy as a novel form of cell death. Ferroptosis has played a significant role in the treatment of GBM. Specifically, research has revealed the key processes of ferroptosis occurrence, including intracellular iron accumulation, reactive oxygen species (ROS) generation, lipid peroxidation, and a decrease in the activity of the antioxidant system. Among them, glutathione peroxidase 4(GPX4) in the cytoplasm and mitochondria, ferroptosis suppressor protein 1 (FSP1) on the plasma membrane, and dihydroorotate dehydrogenase (DHODH) in the mitochondria constitute an antioxidant protection system against ferroptosis. In iron metabolism, nuclear receptor coactivator 4 (NCOA4) can mediate ferritin autophagy to regulate intracellular iron balance based on intracellular iron content. Heme oxygenase1 (HMOX1) catalyzes heme degradation to release iron and regulate ferroptosis. Radiation can trigger ferroptosis by generating ROS, inhibiting the signaling axis of the antioxidant system, depleting glutathione, upregulating acyl-CoA synthase long chain family member 4 (ACSL4), and inducing autophagy. Interestingly, some articles has documented that exposure to low doses of radiation (6 Gy for 24 h or 8 Gy for 4-12 h) can induce the expression of SLC7A11 and GPX4 in breast cancer and lung cancer cells, leading to radiation resistance, while radiation-induced ferroptosis occurs after 48 h. In contrast, high doses of ionizing radiation (20 Gy and 50 Gy) increase lipid peroxidation after 24 h. This suggests that radiation-induced oxidative stress is a double-edged sword that can regulate ferroptosis in both directions, and the ultimate fate of cells after radiation exposure——developing resistance and achieving homeostasis or undergoing ferroptosis——depends on the degree and duration of membrane lipid damage caused by the radiation dose. In addition, during the process of radiotherapy, methods such as inducing iron overload, damaging the antioxidant system, and disrupting mitochondrial function are used to target ferroptosis, thereby enhancing the radiosensitivity of glioblastoma. By promoting the occurrence of ferroptosis in tumor cells as a strategy to improve radiotherapy sensitivity, we can enhance the killing effect of ionizing radiation on tumor cells, thus providing more treatment options for patients with glioblastoma. In this paper, we reviewed ferroptosis and its mechanism, analyzed the molecular mechanism of radiation-induced ferroptosis, and discussed the effective strategies to regulate ferroptosis in enhancing the sensitivity of radiotherapy, with a view to providing an important reference value for improving the current status of glioblastoma treatment.

Comparison of intraoperative balanced salt solution injection or postoperative air filling for superior bullous retinal detachment / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To observe the outcome of intravitreal balanced salt solution(BSS)injection to increase intraocular pressure(IOP)after extrascleral subretinal fluid drainage, then scleral buckling(SB)to treat superior bullous retinal detachment(SBRD), and compare it with the effect of conventional surgery(without any intravitreal filling)and postoperative air filling.METHODS: Retrospective case-control study. A total of 72 patients(73 eyes)who underwent SB for SBRD from January 2018 to December 2022 in ophthalmology department of Xijing Hospital were included. The extrascleral subretinal fluid drainage was performed in all eyes. According to whether intravitreal injection was performed and different injections, patients were divided into three groups: with 24 cases(24 eyes)in the conventional group(no intravitreal injection), 23 cases(23 eyes)in the air group(sterile air was injected after surgery), and 25 cases(26 eyes)in the BSS group(BSS was injected during extrascleral subretinal fluid drainage). All patients were followed up until subretinal fluid was absorbed completely. The average surgery time, postoperative IOP, retinal reattachment rate, subretinal fluid absorption, visual acuity(LogMAR)and major complications were compared.RESULTS: All surgeries were completed successfully. The average surgery time of the conventional group, air group and BSS group were 63.17±13.22, 61.65±15.55 and 57.30±11.70 min, respectively. There had no significant difference among these groups(F=0.825, P=0.443). On the first post-operative day, the average IOP of the conventional group, air group and BSS group were 13.69±2.69, 16.40±2.86 and 18.35±2.88 mmHg, respectively. The average IOP of the air group and the BSS group were significant higher than that of the conventional group(F=17.18, P&#x003C;0.001). Primary reattachment rates were 88%, 96%, and 100%, respectively. The postoperative BCVA was 0.71±0.42, 0.59±0.44, and 0.91±0.50, respectively, which were significantly higher than those before operation(all P&#x003C;0.05), but there was no significant difference among groups(F=3.046, P&#x003E;0.05). The main complications included subretinal hemorrhage in 1 eye from the conventional group and 1 eye from the air group, and a new retinal tear in 1 eye from the air group, resulting in localized retinal detachment.CONCLUSIONS: For SBRD patients with hypotony during SB surgery, intravitreal injection of BSS to properly increase the IOP and then complete the surgery can improve the reattachment rate and reduce postoperative complications. This method is safe and effective for selected SBRD patients.

Silk fibroin methacryloyl hydrogel loaded with Scriptaid regulates polarization of microglia cells / 中国组织工程研究

BACKGROUND:Microglia cells play a major role in maintaining the balance as well as the development and function reconstruction of the central nervous system.As a histone deacetylase inhibitor,Scriptaid can inhibit neuroinflammation and enhance neuroprotection. OBJECTIVE:To investigate the effect of silk fibroin methacryloyl hydrogel loaded with Scriptaid on the behaviors of microglia cells. METHODS:Microglia(BV2 cells)were cultured in medium containing different concentrations of Scriptaid(0,0.1,0.5,1,2,5,10 μmol/L).The optimal concentration of Scriptaid was screened by the CCK-8 assay and live/dead cell staining.Silk fibroin methacryloyl hydrogels loaded with or without Scriptaid were prepared using photocuring.The micromorphology,swelling properties,mechanical properties,slow release properties,and hydrophilicity of the hydrogels were characterized.Microglia(BV2 cells)were inoculated in the subventricular region of 24-well Transwell and cultured in five groups.In the control group,the cell culture medium was added to the lower chamber.In the lipopolysaccharide group,Scriptaid group,hydrogel group,and drug-loaded hydrogel group,cell culture media containing lipopolysaccharide were added into the lower chamber.After 24 hours of lipopolysaccharide intervention,in the Scriptaid group,hydrogel group and drug-loaded hydrogel group,Scriptaid,silk fibroin methacryloyl hydrogel,and silk fibroin methacryloyl hydrogel loaded with Scriptaid were added to the upper chamber,respectively.The culture medium was replaced with ordinary culture medium and continued to culture for 24 hours.The cell viability was detected by CCK-8 assay,and the cell phenotype was detected by immunofluorescence staining of induced nitric oxide synthase and arginase 1. RESULTS AND CONCLUSION:(1)Compared with the group without Scriptaid,the viability and number of BV2 cells were decreased after Scriptaid added.When Scriptaid 2 μmol/L or above was added,the cell viability was lower than the standardized cell viability(70%),and the number of BV2 cells was significantly reduced.Therefore,1 μmol/L Scriptaid was selected to be loaded into the hydrogel.(2)Characterization experiments showed that the addition of Scriptaid did not affect the microscopic morphology,swelling rate of water absorption,compression modulus and hydrophilicity of silk fibroin methacryloyl hydrogel,and silk fibroin methacryloyl hydrogel had slow release performance.(3)The result of CCK-8 assay showed that compared with the control group,silk fibroin methacryloyl hydrogel significantly increased the viability of BV2 cells(P<0.001).(4)Immunofluorescence staining showed that compared with the control group,the expression of inducible nitric oxide synthase was increased in the lipopolysaccharide group(P<0.01);the expression of inducible nitric oxide synthase was decreased(P<0.01)and the expression of arginase 1 was increased(P<0.001)in the drug-loaded hydrogel group;the expression of arginase 1 was increased in the Scriptaid group(P<0.01).(5)The results indicate that Scriptaid-loaded silk fibroin methacryloyl hydrogel is able to promote polarization of microglia to the M2 type after lipopolysaccharide induction.

Comparison study of clinical features between persistent hyperplastic primary vitreous and congenital fibrovascular pupillary membrane / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To analyze the similarities and differences of the clinical features between persistent hyperplastic primary vitreous(PHPV)and congenital fibrovascular pupillary membrane(CFPM).METHODS: Retrospectively analyze the ocular biometric parameters, clinical features and morphological changes in children with PHPV(PHPV group)and CFPM(CFPM group)who received surgery at the department of ophthalmology, Xijing Hospital from March 2006 to December 2021.RESULTS: The study included 56 cases(61 eyes)of PHPV and 24 cases(25 eyes)of CFPM. There were no differences on the gender and age of onset between PHPV and CFPM, and both of them were mainly unilaterally affected, with the ratio of 91% and 96%. Children with PHPV and cataract combined with other complications and ocular developmental abnormalities. CFPM was mainly presented different degrees of blockage and morphological abnormalities of pupillary area. In unilateral cases of PHPV and CFPM, the anterior chamber depth(ACD)of affected eyes was smaller than that of the fellow eyes, and in subgroups with age of operation ≤24mo, the axial length(AL)of affected eyes was smaller than that of the fellow eyes(P&#x003C;0.05). The corneal diameter(CD)of the affected eyes in PHPV group was smaller and the intraocular pressure(IOP)was higher than those of the fellow eyes(all P&#x003C;0.05); while there were no significant differences on CD and IOP between affected eyes and the fellow eyes in CFPM group(P&#x0026;#x003E;0.05). The ACD of affected eyes in PHPV group was significantly smaller than that of CFPM group(P&#x003C;0.05). The fibrovascular membrane in PHPV group was located in the posterior part of the lens and vitreous cavity; while the fibrovascular membrane in CFPM group was located between the iris and the anterior capsule of the lens, rarely involving the lens.CONCLUSION: PHPV and CFPM had the similar clinical features, suggesting that they may belong to the different variants of persistent fetal vasculature(PFV). However, PHPV had a wider range of lesions and more complex conditions.

Progress in research of risk prediction of non-syndromic oral clefts using genetic information / 中华流行病学杂志

Non-syndromic oral cleft (NSOC), a common birth defect, remains to be a critical public health problem in China. In the context of adjustment of childbearing policy for two times in China and the increase of pregnancy at older childbearing age, NSOC risk prediction will provide evidence for high-risk population identification and prenatal counseling. Genome-wide association study and second generation sequencing have identified multiple loci associated with NSOC, facilitating the development of genetic risk prediction of NSOC. Despite the marked progress, risk prediction models of NSOC still faces multiple challenges. This paper summarizes the recent progress in research of NSOC risk prediction models based on the results of extensive literature retrieval to provide some insights for the model development regarding research design, variable selection, model-build strategy and evaluation methods.

Discussion on the relationship between yuan-source points of yang meridians and diseases of fu- organs / 中国针灸

Huangdi Neijing (Yellow Emperor 's Canon of Medicine) establishes the theory that yuan-source points are indicated in zangfu diseases. However, compared with the yuan-source points of yin meridians for the treatment of zang-organ diseases, there is less attention to the indication of yuan-source points of yang meridians for fu-organ diseases, and this statement is even questioned. Collating the early literature and associating with the researches of medical experts, it is found that Nanjing (Classic of Difficult Questions) should be the theoretic origin that yuan-source points of yang meridians are indicated in fu-organ diseases. Regarding the reasons why this theory has not received clinical attention, the three aspects are involved, i.e. the theoretic completion of "he-sea points of three-foot-yang meridians for the diseases of six fu-organs", the limitation of the theory itself, and the lack of literature materials. It is proposed that the exploration on this theory should be deepened in views of the essence of yuan-source points relevant with the characteristic of the wrist-ankle pulse palpation region, acupoint combination, modern technology, etc.

Comparison of HPLC fingerprints and determination of main components of Viticis Fructus from different species / 中国中药杂志

In order to comprehensively evaluate the quality of Viticis Fructus, this study established HPLC fingerprints and evaluated the quality of 24 batches of Viticis Fructus samples from different species by similarity evaluation and multivariate statistical analysis(PCA, HCA, PLS-DA). On this basis, an HPLC method was established to compare the content differences of the main components, including casticin, agnuside, homoorientin, and p-hydroxybenzoic acid. The analysis was performed on the chromatographic column(Waters Symmetry C_(18)) with a gradient mobile phase of acetonitrile(A)-0.05% phosphoric acid solution(B) at the flow rate of 1 mL·min~(-1) and detection wavelength of 258 nm. The column temperature was 30 ℃ and the injection volume was 10 μL. The HPLC fingerprint of 24 batches of Viticis Fructus samples was established with 21 common peaks, and nine peaks were identified. Similarity analysis was carried out based on chromatographic data of 24 batches of chromatographic data of Viticis Fructus, and the results showed that except for DYMJ-16, the similarity of Vitex trifolia var. simplicifolia was ≥0.900, while that of V. trifolia was ≤0.864. In addition, the similarity analysis of two different species showed that the similarity of 16 batches of V. trifolia var. simplicifolia was 0.894-0.997 and that of the eight batches of V. trifolia was between 0.990 and 0.997. The results showed that the similarity of fingerprints of these two species was different, but the similarity between the same species was good. The results of the three multivariate statistical analyses were consistent, which could distinguish the two different species. The VIP analysis results of PLS-DA showed that casticin and agnuside contributed the most to the distinction. The content determination results showed that there was no significant difference in the content of homoorientin and p-hydroxybenzoic acid in Viticis Fructus from different species, but the content of casticin and agnuside was significantly different in different species(P<0.01). The content of casticin was higher in V. trifolia var. simplicifolia, while agnuside was higher in V. trifolia. The findings of this study show that there are differences in fingerprint similarity and component content of Viticis Fructus from different species, which can provide references for the in-depth study of the quality and clinical application of Viticis Fructus.

Effects of Xihuang Pills on proliferation and apoptosis of prostate cancer LNCaP cells based on AR/m TOR signaling pathway / 中国中药杂志

Based on the androgen receptor(AR)/mammalian target of rapamycin(mTOR)signaling pathway, the effects of Xihuang Pills-medicated serum on the proliferation and apoptosis of prostate cancer LNCaP cells were investigated. The drug-containing serum of SD rats was prepared by intragastric administration of Xihuang Pills suspension. The effects of low-, medium-, and high-dose Xihuang Pills-containing serum on the in vitro proliferation of LNCaP cells were detected by cell counting kit-8(CCK-8). Flow cytometry was used to detect the apoptosis level of LNCaP cells after intervention with different concentrations of Xihuang Pills. Protein expression of cleaved cysteinyl aspartate-specific proteinase caspase-3(cleaved caspase-3), B-cell lymphoma-2(Bcl-2), and AR as well as the phosphorylation level of mTOR protein were detected by Western blot. The results showed that compared with the blank serum, the drug-medicated serum could blunt the activity of LNCaP cells. Low-, medium-, and high-dose Xihuang Pills-containing serum could significantly increase the cell apoptosis rate, increase the expression of cleaved caspase-3 protein, decrease the expression of Bcl-2 protein, reduce the expression of AR protein, and down-regulate the level of phosphorylated mTOR(p-mTOR). To study the effect of Xihuang Pills on the growth of LNCaP cells in vivo, different doses of Xihuang Pills were used to intervene in the subcutaneous graft model in nude mice inoculated with LNCaP cells. The expression levels of AR, mTOR, p-mTOR, Bcl-2, and cleaved caspase-3 were detected by Western blot. The results showed that the volumes of subcutaneous graft tumor in the low-dose, medium-dose, and high-dose Xihuang Pills groups significantly decreased compared with that in the model group. The weight of subcutaneous transplanted tumor in each group with drug intervention was significantly lower than that in the model group. Compared with the model group, the low-dose, medium-dose, and high-dose Xihuang Pills groups showed increased cleaved caspase-3 protein expression, decreased Bcl-2 and AR protein expression, and reduced p-mTOR protein expression. Further experiments showed that AR agonist R1881 could block the anti-proliferation and pro-apoptotic effects of Xihuang Pills. The mechanism of Xihuang Pills against prostate cancer is related to the inhibition of the AR/mTOR signaling pathway, inhibition of LNCaP cell proliferation, and induction of apoptosis in cancer cells.

Second allogeneic hematopoietic stem cell transplantation with reduced-intensity conditioning and donor changes in relapsed hematological malignancies after the first allogeneic transplant / 中华血液学杂志

Objective: The purpose of this study was to assess the safety and efficacy of a second allogeneic hematopoietic stem cell transplantation (allo-HSCT) with reduced-intensity conditioning (RIC) in patients with hematological malignancies who had relapsed after the first allo-HSCT. Methods: Between April 2018 and June 2021, 44 patients with hematological malignancies (B-ALL 23, T-ALL/T-LBL 4, AML15, and MDS 2) were enrolled and retrospectively examined. Unrelated donors (n=12) or haploidentical donors (n=32) were used. Donors were replaced in all patients for the second allo-HSCT. Hematological and immunological germline predisposition genes and hematopoietic and immune function tests were used to select the best-related donor. Total body irradiation (TBI) /fludarabine (FLU) -based (n=38), busulfan (BU) /FLU-based (n=4), total marrow irradiation (TMI) /FLU-based (n=1), and BU/cladribine-based (n=1) were the RIC regimens used. For graft versus host disease (GVHD) prevention, cyclosporine, mycophenolate mofetil, short-term methotrexate, and ATG were used. Eighteen (40.9%) of 44 patients with gene variations for which targeted medications are available underwent post-transplant maintenance therapy. Results: The median age was 25 years old (range: 7-55). The median interval between the first and second HSCT was 19.5 months (range: 6-77). Before the second allo-HSCT, 33 (75%) of the patients were in complete remission (CR), whereas 11 (25%) were not. All patients had long-term engraftment. The grade Ⅱ-Ⅳ GVHD and severe acute GVHD rates were 20.5% and 9.1%, respectively. Chronic GVHD was found in 20.5% of limited patterns and 22.7% of severe patterns. CMV and EBV reactivation rates were 29.5% and 6.8%, respectively. Hemorrhage cystitis occurred in 15.9% of cases, grade Ⅰ or Ⅱ. The 1-yr disease-free survival (DFS), overall survival (OS), and cumulative recurrence incidence (RI) rates of all patients were 72.5% (95% CI, 54.5%-84.3%), 80.6% (95% CI, 63.4%-90.3%), and 25.1% (95% CI, 13.7%-43.2%), respectively, with a median follow-up of 14 (2-39) months. There were eight deaths (seven relapses and one infection). The rate of non-relapse mortality (NRM) was only 2.3%. The CR patients' 1-yr RI rate was significantly lower than the NR patients (16.8% vs 48.1%, P=0.026). The DFS rate in CR patients was greater than in NR patients, although there was no statistical difference (79.9% vs 51.9%, P=0.072). Univariate analysis revealed that CR before the second allo-HSCT was an important prognostic factor. Conclusion: With our RIC regimens, donor change, and post-transplant maintenance therapy, the second allo-HSCT in relapsed hematological malignancies after the first allo-HSCT is a safe and effective treatment with high OS and DFS and low NRM and relapse rate. The most important factor influencing the prognosis of the second allo-HSCT is the patient's illness condition before the transplant.

Mechanistic study of Cyclosporin A in alleviating cardiac injury in model mice with acute pancreatitis / 中国免疫学杂志

Objective:To observe whether Cyclosporin A(CSA)treatment may protect acute pancreatitis(AP)mice and related cardiac injury by consequent attenuation of systemic inflammation via regulating macrophage polarization.Methods:RAW264.7 cells were used for in vitro experiments,stimulated with 0,5,10 and 20 nmol/L CSA for 24 hours,and M2 markers were detected by flow cytometry.C57BL/6J mice were used for experiments in vivo.The mice were randomly divided into 3 groups(n=15):control group,AP model group(intraperitoneal injection of L-arginine)and AP+CSA(20 mg/kg)group,CSA was administered in a pre-treated manner.ELISA kit was used to detect the indexes of pancreatic and myocardial injury in mice;HE staining was used to detect the pathological changes of pancreas and heart tissue;TUNEL method was used to detect apoptosis in tissue sections;CETSA was used to determine the relationship between CSA and PKM;The expressions of PKM2,HIF1α,p-STAT1 and p-STAT6 were detected by Western blot.Results:CSA increased the number of M2 macrophages and decreased the number of M1 macrophages in a dose-dependent manner in vitro.CSA pretreatment significantly improved pancreatic structure and myocardial injury in AP mice,decreased pancreatic histopathological score and serum amylase,lipase,TNF-α,CK-MB,LDH and cTnT levels.CSA pretreatment significantly reduced the number of TUNEL positive cells in myocardium of AP mice.Flow cytometry analysis showed that CSA pretreatment significantly inhibited the proportion of CD11c+F4/80+ and promoted the ratio of CD206+F4/80+ in AP induced myocardial macrophages.CETSA analysis showed that PKM2 was the target of CSA.Conclusion:CSA can significantly improve the severity of L-arginine-induced cardiac damage in AP model mice,and its mechanism of action is related to increasing the number of M2 macro-phages and inhibiting the production of proinflammatory cytokines.

Progress in clinical diagnosis and treatment of tracheoesophageal fistulas / 解放军医学杂志

Tracheoesophageal fistulas(TEF)is a kind of pathological fistula between trachea or bronchus and esophagus,including tracheoesophageal fistula and bronchoesophageal fistula.In clinical practice,tracheoesophageal-fistula is more often seen.There are many pathogenesis of TEF,which could cause serious complications.It is a disease which has serious impact on the quality of life,difficult to treat and high mortality.There are many methods to treat TEF,but the therapeutic effect is poor.There is an urgent need for new treatment methods to TEF.The detection of nasogastric tube retention by chest and abdominal X-ray is the gold standard for diagnosis.The main treatment methods include surgical treatment,stent implantation,local biological glue blocking,stem cell therapy and so on.In order to provide reference for relevant clinical diagnosis and treatment of TEF,this article reviews the main research progress in the diagnosis and treatment of TEF in recent years.

Role of sclerostin in the occurrence and development of uveal melanoma and its biological mechanism / 中华实验眼科杂志

Objective:To investigate the effects of sclerostin (SOST) and WNT/CTNNB1 signaling pathway on the cell cycle, migration and invasion of human uveal melanoma (UM) cells and its related mechanism.Methods:UM tissues from 20 cases of epithelioid UM and 16 cases of spindle cell type UM were collected.The contents of SOST, Wnt-1 and Catenin beta-1 proteins in the collected tissues were detected by immunohistochemical staining.Three human UM tissue derived cell lines OCM-1 (primary spindle cell type), Mum-2B (metastatic epithelioid) and Mum-2C (metastatic spindle cell type) were selected and divided into three groups, blank control group not transfected, empty vector group transfected with SOST negative control vector and SOST siRNA group transfected with SOST siRNA.After 24-hour transfection, the mRNA and protein expression levels of SOST, CTNNB1, WNT protein family 1 (WNT1), CCND1, matrix metalloproteinase (MMP)2 and MMP9 were detected by real-time fluorescence quantitative PCR and Western blot, respectively.The invasion and migration ability of the transfected cells were measured by transwell method, and the cell cycle distribution was detected by flow cytometry.Another 9 female BALB/c nude mice were selected and randomized into OCM-1 group, OCM-1 empty vector group and SOST shRNA group, inoculated with OCM-1 without lentivirus infection, OCM-1 with blank lentivirus infection and OCM-1 with SOST shRNA lentivirus infection, respectively.Six weeks after inoculation, the in situ formation of tumor was observed.The interaction between SOST and low density lipoprotein receptor related protein(LRP)-5/6 in OCM-1 cells was explored by co-immunoprecipitation assay.The study protocol was approved by the Ethics Committee of Tianjin Medical University Eye Hospital (2018KY[L]-20).Results:Immunohistochemical staining results showed that the SOST expression level was higher and the expression levels of Wnt-1 and Catenin beta-1 were lower in spindle cell type UM tissues than in epithelioid UM tissues, and the differences were all statistically significant (all at P<0.01). The real-time fluorescence quantitative PCR results showed that the relative expression of SOST mRNA was significantly lower and the relative expressions of CCND1, WNT1 and MMP9 mRNA were significantly higher in SOST siRNA groups than in corresponding empty vector groups in the three cell lines (all at P<0.05). In OCM-1 and Mum-2C cell lines, the relative expressions of CTNNB1 mRNA were significantly higher in SOST siRNA groups than in empty vector groups (all at P<0.01). Western blot results showed that the relative expression of SOST protein was significantly lower and the relative expressions of Wnt-1, Catenin beta-1, cyclin-D1, MMP2 and MMP9 proteins were significantly higher in SOST siRNA groups than in empty vector groups (all at P<0.01). Transwell assay showed that the cell invasion and migration ability of SOST siRNA group was significantly higher than that of blank control group and empty vector group in the three cell lines (all at P<0.01). Flow cytometry showed that the proportion of G1-phase cells and the G1/S-phase ratio were significantly lower in SOST siRNA group than in blank control groups and empty vector groups (all at P<0.01). The eyeball volume of OCM-1 group, OCM-1 empty vector group and SOST shRNA group was (42.7±4.6), (49.0±22.9) and (135.2±32.7)mm 3, respectively, showing a significant overall difference ( F=19.963, P<0.01). The eyeball volume of SOST shRNA group was larger than that of OCM-1 group and OCM-1 empty vector group, and the differences were statistically significant (both at P<0.05). Co-immunoprecipitation results showed that SOST could interact with LRP-5 and LRP-6 by binding to them, respectively. Conclusions:Silencing SOST can promote the invasion and migration of UM cells, and increase the proportion of UM cells in the division phase.Silencing SOST can promote tumor growth in eyes of nude mice.SOST may play this function by interacting with the membrane receptor LRP-5/LRP-6 and then regulating the WNT/CTNNB1 signal pathway.

Prediction of Ki-67 expression level in mass breast cancers using multi-modal ultrasound features / 中华超声影像学杂志

Objective:To predict the Ki-67 expression grading in patients with mass breast cancer based on multimodal ultrasound features to aid clinical diagnosis and treatment.Methods:Ninety-three female patients (93 masses in total) with breast cancers confirmed by pathological examination were retrospectively included in the Second Affiliated Hospital of Harbin Medical University from September 2017 to September 2020. According to the immunohistochemical results, the patients were divided into Ki-67 high expression group (55 cases) and Ki-67 low expression group (38 cases). The qualitative and quantitative features from two-dimensional gray-scale ultrasound, color Doppler flow imaging (CDFI), shear wave elastography (SWE) and contrast-enhanced ultrasound (CEUS) images of all breast masses were retrospectively analyzed, differential features were analyzed based on logistic regression algorithm. ROC curves and Kappa test were used for the evaluation of diagnosis.Results:The univariate analysis revealed statistically significant differences between the two groups for conventional ultrasound features (size, shape, margins), SWE features (stiff rim sign, Eratio), and CEUS features (perfusion defect, IMAX) (all P<0.05). In the multiple logistic regression analysis, the margins, stiff rim sign, and perfusion defect were the independent factors for predicting the Ki-67 expression (all P<0.05). The performance of the predictive model was 0.882 (95%confidence interval of 0.798-0.940, P<0.05) with the sensitivity of 0.818 and specificity of 0.790. Conclusions:A preliminary analysis of the relationship between multi-modal ultrasound features and Ki-67 expression grading in mass breast cancers was performed based on logistic regression algorithm to provide more imaging information for clinical treatment and prognosis assessment.

Combining bi-modal imaging and immunohistochemical indicators to predict axillary lymph nodes status in cN 1 breast cancer patients receiving neoadjuvant chemotherapy / 中华超声影像学杂志

Objective:To investigate whether ultrasound features, mammographic features and immunohistochemical indicators show any association with rates of axillary pathologic complete response(pCR) in cN 1 breast cancer patients receiving neoadjuvant chemotherapy(NAC), and to construct prediction models of axillary pCR to predict axillary lymph nodes (ALN) status, so as to select suitable patients for less invasive axillary surgery after NAC. Methods:This retrospective study evaluated 134 consecutive cN 1 breast cancer patients with ALN metastasis who underwent NAC in the Second Affiliated Hospital and Tumor Hospital of Harbin Medical University from July 2020 to July 2022. According to the pathological results of ALN surgery after NAC, the cases were divided into pCR and non pathologic complete respose(npCR) groups. The ultrasound images, mammographic images and immunohistochemical indicators of the two groups were compared. In terms of logistic regression algorithm, the model A(the ultrasound model), the model B(the ultrasound combined with mammography model), the model C(the ultrasound combined with immunohistochemistry model) and the model D(the ultrasound combined with mammography and immunohistochemistry model) were respectively established for predicting the pathological state of axillary lymph nodes in breast cancer patients, ROC curves were plotted to evaluate the performance of the models, and the diagnostic efficiency of different models was compared by Delong′s test. The model with the best predictive performance was shown in a nomogram. Results:①The P values between two groups of the short diameter of ALN, the ratio of long/short diameter of ALN, fatty hilum and central hilar vascularity, mammographic spiculation, estrogen receptor(ER), progesterone receptor(PR), human epidermal growth factor receptor 2(HER2) were <0.05 by the t test and χ 2 test analysis. ②The ratio of long/short diameter and fatty hilum in the model A were independent factors for predicting the pathological status of ALN after NAC. The independent predictors of model B and Model C were respectively added with mammographic spiculation and immunohistochemical indicators (ER, PR) on the basis of model A. In the model D, the ratio of long/short diameter, short diameter, fatty hilum, mammographic spiculation, and immunohistochemical indicators (ER, PR) remained significant independent predictors associated with axillary pCR. ③The area under ROC curve (AUC) of the model A, B, C, D was 0.78, 0.84, 0.84 and 0.89, respectively. The sensitivity was 0.71, 0.80, 0.78 and 0.86, the specificity was 0.76, 0.74, 0.76 and 0.80, and the accuracy was 0.73, 0.76, 0.77 and 0.83, respectively. ④Delong′s test showed the model D had an improved AUC of 0.89(0.89 vs 0.78, 0.84, 0.84, all P<0.05). Conclusions:The prediction models combining bi-modal imaging and immunohistochemical indicators show good prediction ability and can provide reference for selecting suitable patients for less invasive axillary surgery after NAC.

Assessment of conventional magnetic resonance morphological measurements in diagnosis and differential diagnosis of progressive supranuclear paralysis / 中华老年医学杂志

Objective:To study the features as well as the diagnosis and differential diagnosis values by conventional MRI morphometrics in different clinical subtypes of progressive supranuclear palsy(PSP).Methods:Forty five patients with PSP were included, comprising three PSP subtypes: 15 cases of Richardson's syndrome(PSP-RS), 15 cases of Parkinson's syndrome(PSP-P)and 15 cases of progressive frozen gait(PSP-PFG). In addition, three control groups were established: 15 cases of multiple system atrophy-Parkinson's syndrome(MSA-P), 30 cases of primary Parkinson's disease(PD)and 40 healthy controls(HC). Midbrain area-to-Pons area ratio(M/P), Magnetic Resonance Parkinsonism Index(MRPI, MRPI2.0), width ratio of middle cerebellar peduncle to superior cerebellar peduncle(MCP/SCP), Midbrain-to-Pons ratio(MTPR), Angle of cerebral peduncle(A cp), third ventricle width/frontal horns width ratio(V 3rd/FH), and Humming bird sign rating scale(HBS-RS)scores were calculated.Diagnostic sensitivity and specificity were performed by ROC curve to assess the accuracy of these imaging indicators in the diagnosis and differential diagnosis of PSP and its subtypes. Results:The MRPI, MRPI2.0, MCP/SCP and HBS-RS scores were significantly higher in PSP group than in other control groups( H=69.351, 66.776, 33.926 and 84.694, all P<0.05), while M/P and MTPR were significantly lower in PSP group than in other control groups(H=60.101 and 77.276, all P<0.05). PSP group also had higher V 3rd/FH compared with PD or HC group( F=17.168, P<0.05), but not with MSA-P group( Z=-1.602, P>0.05). The above differences also existed between each PSP subgroup and control groups.Among PSP subgroups, PSP-PFG subgroup had a larger A cp than did PSP-RS( Z=-2.510, P<0.05), and had higher HBS-RS score than did PSP-P group( Z=-2.380, P<0.05). No significant differences in other MRI morphometric indexes were identified among PSP subtypes.The M/P, MRPI, MTPR, MRPI2.0, HBS-RS score showed good accuracy in diagnosing PSP and its each subgroup, with HBS-RS score being the most accurate indicator, when the cutoff value was 2, the AUC values were all higher than 0.99, and the sensitivity and specificity were all above 90%.PSP and its subtypes were best distinguished from MSA-P by MRPI, when the cutoff value was 9.94, the AUC values were all higher than 0.90, with the sensitivity of 100% and specificity of 86.67%.PSP and its subtypes were best distinguished from PD by MTPR, AUC values were all above 0.95, with slightly different cutoff values.Almost all the morphological measurement parameters failed to show significant sensitivity and specificity in discriminating subtypes of PSP.The sensitivity and specificity of almost all MRI morphometry indicators in differentiating different subtypes of PSP are not high. Conclusions:MRI morphometrics have a high value both in the diagnosis of PSP and its subtypes, and also in specific application fields.MRI morphometrics have a limited value in discriminating PSP subtypes.

Herbal Textual Research on Gei Herba and Its Pharmacological Effect on Cardiovascular and Cerebrovascular Diseases / 中国实验方剂学杂志

Gei Herba is a traditional folk herbal medicine with a variety of functions such as replenishing Qi and invigorating spleen， tonifying blood and nourishing Yin， moistening lung and resolving phlegm， activating blood and alleviating edema， moving Qi， and activating blood. The reports about the pharmacological effects of this herbal medicine have been increasing in recent years. By reviewing the ancient and modern literature about Gei Herba， we systematically organized the name， original plants， nature， taste， and functions of this herbal medicine， and summarized the modern pharmacological studies and clinical applications of Gei Herba in cardiovascular and cerebrovascular diseases. Gei Herba was first recorded in the name of "Dijiao" in the Geng Xin Yu Ce（《庚辛玉册》） written in the Ming Dynasty. It is derived from Geum japonicum var. chinense （Rosaceae） and sometimes confused with Adina rubella （Rubiaceae）. This medicine had numerous synonyms in the local materia medica books. Gei Herba is widely distributed and harvested in summer and autumn， with the dried whole grass used as medicine. The historical records of the nature， taste， meridian tropism， main functions， and indications of Gei Herba are not consistent. It is generally believed that Gei Herba is pungent， bitter， sweet， cool， and has tropism to the liver， spleen， and lung meridians. Based on the effects of tonifying Qi， activating blood， and nourishing Yin， modern pharmacological studies have reported that the extracts of Gei Herba and the tannin phenolic acid compounds and triterpenoids isolated from Gei Herba have therapeutic effects on cardiovascular and cerebrovascular diseases such as hypertension， myocardial ischemia， cerebral ischemia， and vascular dementia. This study provides a reference for discovering the clinical advantages of Gei Herba and developing new drugs.

Construction and optimization of a novel plasma membrane model for membrane affinity assay based on polydiacetylene vesicles / 药学学报

Most drugs need to interact with cell membrane to reach the biological target, so that membrane affinity assay is an important early screening step in drug discovery. However, at present, the traditional oil-water distribution method is still used, a new, simple and accurate method for membrane affinity assay is urgently needed. In this study, according to the colorimetric principle, a new assay model based on polydiacetylene vesicles was optimized through a series of experiments including different concentrations of vesicle solution, temperature, or pH reaction environment. On this basis, tetracaine hydrochloride, 2-methylimidazole and histamine were used as model drugs to measure the membrane affinity constants and verify the between-batch precision of the optimized assay model (relative standard deviation less than 5%). In addition, polydiacetylene vesicles were stable for up to 180 days, demonstrating the potential application of the assay model. This strategy is simple, stable, reliable, with high reproducibility, low cost and easy to promote, which provided a new tool and a new direction for the high-throughput assay of membrane affinity.

Effect of obesity induced by high-fat diet and adipose exosomes on knee post-traumatic osteoarthritis caused by destabilization of medial meniscus surgery in young mice / 药学学报

Obesity is an important risk factor related to osteoarthritis, but it′s role in post-traumatic osteoarthritis on young people need to further study. The internal mechanism except the mechanical loading may be associated with adipose exosomes. To examine the effect of obesity induced by high fat diet and adipose exosomes on knee post-traumatic osteoarthritis caused by destabilization of medial meniscus (DMM) surgery in young mice, 20 6-week-old C57BL/6J mice were randomly assigned to the control diet group (CD, n = 5), the DMM group (n = 5), the high fat diet group (HFD, n = 5) and the HFD plus DMM group (HFD+DMM, n = 5). The CD and DMM group were fed with a control diet, and the HFD and HFD+DMM group were fed with a high fat diet. We did the DMM surgery and the sham surgery on the mice when it was 10 weeks old. Extract obese and normal adipose exosomes, identify exosomes in vitro, and proceed fluorescence imaging in vivo using DiR staining. DMM+HFD-Exo group and DMM+CD-Exo group were injected the exosomes from the tail vain once a week (100 μL per shot with a concentration of 1 μg·μL-1). Second, 15 6-week-old C57BL/6J mice were randomly assigned to the DMM group (n = 5), the DMM plus obese adipose exosomes group (DMM+HFD-Exo, n = 5), and the DMM plus control diet adipose exosomes group (DMM+CD-Exo, n = 5). Animal welfare and experimental process are in accordance with the regulations of the Experimental Animal Ethics Committee of Nanjing University (IACUC-D2204005). All mice were sacrificed at the age of 18 weeks, the knee joints of the mice were harvested and fixed. We used micro CT to examine the samples and measured the bone volume/tissue volume, trabecular thickness, trabecular number and trabecular separation. Then the samples were decalcified and embedded in paraffin, and 4 μm thickness sections were stained with H&E and safranin O/fast green to observe the histological changes of the knee joint. The results showed compared with the control diet group, high fat diet induced obesity can aggravate the pathological changes of the post-traumatic osteoarthritis caused by DMM surgery, which shows in having a higher Mankin score. The surface of knee articular cartilage in the HFD+DMM group was rough, and the subchondral bone has an increase in bone sclerosis. Compared with the DMM group, obese adipose exosomes can exacerbate the pathological changes of the knee articular cartilage, while not influencing the subchondral bone. In conclusion, high fat diet induced obesity can aggravate the post-traumatic osteoarthritis caused by DMM surgery in young mice. The obese adipose exosomes mainly affect the surface of the knee articular cartilage.