Objective To investigate the relationship between acute radiation proctitis and radiation dose,volume as well as radiation time,in the process of intensity-modulated radiation therapy (IMRT) for the cervical cancer patients.Methods A total of 51 patients with locally advanced cervical cancer were enrolled from January 2011 to December 2013.Those patients were then classified into grade 1 to 4 groups,according to the RTOG/EORTCtoxicity grading standard.The exposure dose volume and the average dose of rectum under the standard plan were evaluated with dose-volume histogram (DVH).The ANOVA test was used for analyzing Dmax,D mean,D1 cm3,D2cm3,D40 and V40 values of rectum and the average exposure dose of rectum.Results The average time of acute radiation proctitis with clinical symptoms was (23.06 ± 12.01) d after radiotherapy.Dmaxvalues of rectum in grade 2 group was lower than those in grade 3 and 4 groups (F =5.268,P ＜ 0.05).Moreover,D1 cm3 and D2 cm3 values of rectum in grade 1 and 2 groups were also lower than those in grade 3 and 4 groups (F =4.893,4.406,P ＜ 0.05).There was no statistically significant difference between D40 and V40 values.Conclusions The acute radiation proctitis could be frequently found around 20 days during the IMRT for cervical cancer patients.Mild and moderate acute radiation proctitis are more common,while severe acute radiation proctitis is rare.Minimizing Dmax,D1 cm3 and D2 cm3 values of rectum might reduce the incidence of severe acute radiation proctitis in cervical cancer patients receiving IMRT.

Objective: To evaluate the efficacy, toxicity and cosmetic outcomes of hypofractionated simultaneous integrated boost intensity-modulated radiotherapy (IMRT-SIB) after breast conservative surgery (BCS) for early breast cancer patients. Methods: A total of 76 patients with stage TisT_1-2N₀M₀ breast cancer treated with BCS were enrolled in the analysis. The patients who underwent breast radiotherapy without regional lymph node irridiation and hypo fractionated IMRT/VMAT were used. All patients received whole breast IMRT/VMAT with tumor bed SIB. The doses delivered to the whole breast was 42.4 Gy in 16 fractions, and the dose delivered to tumor bed for SIB was 49.6 Gy in 16 fractions. Cosmetic evaluation was based on the Harvard system. Acute and late toxicities were scored according to CACAT version 3.0. Survival and recurrence rates were calculated by Kaplan-Meier method. The univariate and multivariate analysis were conducted with logistic regression. Results: The median follow-up was 29 months (range 16-40 months). The follow-up rate was 100%. The 1-, 2-and 3-year overall survival rates were 100%. No recurrence or metastasis was observed in this study. The incidence of grade 1 acute skin toxicity was 68.4%, grade 2 was 7.9%. The late skin toxicity of grade 1 was 13.1%, grade 2 was 2.6%.In all, 82.4% of patients had excellent and good cosmetic outcome. The Mean dose of the tumor bed was predictive factor for grade 2 dermatitis. Conclusion: The efficacy, cosmetic effect, the acute and late treatment-related toxicity of hypofractionated IMRT/VMAT-SIB in patients with early breast cancer following BCS might be acceptable. A longer follow-up is needed to define the efficacy on outcomes. Trial registration Chinese clinical trial registry, ChiCTR1800016287

Ischemia reperfusion (I/R) injury is caused by ischemic shock, cardiac arrest, resection and transplantation, and its mechanism is closely related to calcium overload. Mitochondrial calcium uniporter (MCU) is a highly selective calcium channel located in the mitochondrial inner membrane (IMM), mediating calcium ions into the mitochondrial matrix. The MCU complex with the MCU as the core plays an important role in maintaining calcium ion homeostasis and alleviating I/R injury in the heart, kidney, brain, liver and other organs. The mitochondria associated endoplasmic reticulum membranes (MAMs) is a key protein in this process.

BACKGROUNDThe present experimental data have showed that the function of kinase suppressor of Ras (KSR) is mainly as a scaffold protein that coordinates the assembly of a multiprotein complex containing MAPK and its upstream regulators. But whether KSR has kinase activity is still the point of argument until now. The aim of this study is to construct eukaryotic expression vectors of carboxyl terminus and amino terminus of KSR and to detect their expression in 293T cell line.

METHODSN-KSR and C-KSR were amplified by PCR. The eukaryotic expression vectors of pCMV-Tag2b-N-KSR and pCMV-Tag2b-C-KSR were constructed by gene recombination technique and the recombinant plasmids were verified by restriction enzyme analysis and sequencing. Then positive clones were transfected into 293T cell line. Expression of target proteins was analyzed by Western blot.

RESULTSThe sequences and open read frames of the two vectors were both completely concordant with experiment design. The target proteins could be observed in transfected 293T cells by Western blot.

CONCLUSIONSEukaryotic expression vectors of pCMV-Tag2b-N-KSR and pCMV-Tag2b-C-KSR are successfully constructed, and they can be expressed in 293T cells. It provides an experimental base for further research work.

This study was performed to investigate the features of HBV-specific CD8+T cell reactivity in patients with chronic hepatitis B(CHB),HBV-induced liver cirrhosis(LC)or hepatocellular carcinoma(HCC).A total of 124 CHB patients,36 LC patients,and 114 HCC patients were enrolled in this study.The reactive HBV-specific CD8+T cells in peripheral blood were enumerated using an innovative ELISPOT system.In addition,19 CHB patients and 20 HCC patients were longitudinally monitored with an interval of 3-5 months.Data showed that the numbers of reactive HBV-specific CD8+T cells in CHB group were not significantly different from that in LC group,but obviously lower than that in HCC group(P=0.009 9),especially HBsAg-,HBpol-and HBe/cAg-specific CD8+T cells.In CHB group,the patients with normal ALT level,AST level,or low HBV-DNA load showed significantly more reactive HBV-specific CD8+T cells than the patients with abnormal ALT level,abnormal AST level,or high HBV-DNA load.Furthermore,the duration of NUCs treatment had an impact on the HBV-specific CD8+T cell reactivity in CHB patients,while different NUCs at the same treatment duration did not bring different reactivity of HBV-specific T cells.In LC group,the HBeAg-positive patients presented much more reactive HBV-specific CD8+T cells than the HBeAg-negative patients did.In HCC group,the numbers of reactive HBV-specific CD8+T cells in the patients with normal AFP level or normal DCP level were significantly higher than that in the patients with abnormal AFP level or abnormal DCP level.Longitudinal monitoring results showed that HBV-specific CD8+T cell reactivity displayed a slow upward trend in the CHB patients undergoing NUCs treatment,and an obvious increasing in the HCC patients undergoing combined treatment of targeted drugs and immunotherapy.Taken together,the features of HBV-specific CD8+T cell reactivity are distinct among the CHB,LC and HCC patients,and are influenced by virological indicators,tumor markers and treatment regimens.Therefore,more attention should be paid to the changes of HBV-specific CD8+T cell reactivity during clinical treatment.

Objective:To establish HPLC chromatogram for Aspongopus; To determine 8 nucleoside components of uracil, adenine, uridine, uric acid, hypoxanthine, adenosine, xanthine and canine quinolinic acid; To provide reference for quality control and evaluation.Methods:The Agilent ZORBAX SB-Aq chromatographic column (4.6 mm×250 mm, 5 μm) was used for gradient elution with mobile phases consisting of a methanol (A) and 0.05% phosphoric acid (B). The column temperature was 25 ℃, the flow rate was 0.8 ml/min, and the detection wavelength was 254 nm. HPLC chromatograms for Aspongopus were established and the contents of 8 components were determined.Results:The characteristic chromatogram of 15 batches aspongopus herbs was established. A total of 10 common characteristic peaks were identified and 8 were identified. The similarity between the characteristic chromatogram of samples and the control chromatogram was 0.969-0.997. The content determination showed that the linear range of uracil, adenine, urin, uric acid, hypoxanthine, adenosine, xanthine and xanuric acid was among 0.002 0-0.644 0, 0.001 4-0.448 0, 0.001 0-1.257 0, 0.005 4-6.221 0, 0.001 0-0.724 0, 0.001 0-0.644 0, 0.002 0-1.113 0, 0.003 8-2.059 0 μg, respectively, with a good linear relationship ( r≥0.999); the repeatability and stability of RSD were <2.0%, and the average sampling recovery rate was between 99.36% and 103.40%. Conclusion:The characteristic chromatogram and content determination method established in this study are simple, reliable, reproducible and accurate, and can be used for the qualitative and quantitative analysis of Aspongopus and can provide a reference for the quality evaluation method of the Aspongopus.

Objective To investigate the efficacy and safety of sofosbuvir/velpatasivr/voxilaprevir(SOF/VEL/VOX)in patients with HCV infection experiencing failure in previous direct-acting antiviral agent(DAA)therapy.Methods A retrospective analysis was performed for the chronic hepatitis C patients who experienced failure in previous DAA antiviral therapy and were treated with SOF/VEL/VOX(400 mg/100 mg/100 mg/tablet,1 tablet/day)for 12 weeks in Nanjing Second Hospital,Wuxi Fifth People's Hospital,and The Third People's Hospital of Zhenjiang from June 2020 to June 2023.Sustained virological response at 12 weeks(SVR12)was observed after the end of treatment,and the changes in biochemical parameters and the incidence rate of adverse reactions were assessed to evaluate drug safety.The paired t-test was used for comparison of continuous data between two groups.Results A total of 36 patients were enrolled,among whom there were 27 non-liver cirrhosis patients and 9 patients with compensated liver cirrhosis,and 4 patients experienced failure in the previous two or more sessions of DAA therapy.Two patients were lost to follow-up after treatment,and the remaining 34 patients(34/36,94.4%)achieved SVR12.Among the 36 patients enrolled,the most common adverse events were pruritus,nausea,fatigue,and headache,and one patient(2.78%)experienced serious adverse events;there were no adverse events that resulted in the discontinuation of therapeutic agents or the death of patients.Conclusion For chronic hepatitis C patients who experience failure in previous DAA therapy,SOF/VEL/VOX salvage therapy has a relatively high rate of SVR12,with good tolerability and safety.

Objective:To explore the curative effects of Nice knot fixation on tuberosity healing in hemiarthroplasty for complex proximal humeral fractures.Methods:A retrospective analysis was conducted of the eligible 32 complex proximal humeral fractures which had been treated at Department of Trauma and Orthopedics, Peking University People's Hospital between May 1, 2016 and May 1, 2019. Nice knot fixation was used to repair greater and lesser tuberosities in hemiarthroplasty for all the patients. There were 6 males and 26 females, aged from 60 to 90 years (mean, 74.9 years). By the Neer classification, there were 4 three-part fractures combined with dislocation, 20 four-part fractures, and 8 four-part fractures combined with dislocation. Shoulder joint X-rays were taken at postoperative 1, 2, 3, 6, and 12 months at the outpatient clinic to evaluate the patients' shoulder joint mobility, visual analog scale (VAS) pain score and Constant-Murley shoulder score. Tuberosity healing was assessed based on the X-rays and related complications were recorded.Results:The 32 patients received complete follow-up for 12 to 25 months (average, 17.82 months). At the 12-month follow-up, their shoulder flexion averaged 131.3° (from 80° to 155°), abduction 126.9° (from 80° to 155°), external rotation 48.4° (from 30° to 60°), internal rotation the L2 level, VAS pain score 0.9 (from 0 to 5), and Constant-Murley score 83.4 (from 58 to 96). The rate of patient satisfaction was 87.5%(28/32). Tuberosity-related complications were observed in 6 cases with an incidence of 18.8%. Complications like infection and prosthetic loosening were found in none of the patients.Conclusion:In hemiarthroplasty for complex proximal humeral fractures, application of Nice knot to fixate greater and lesser tuberosities can lead to rigid fixation, definite curative effects and a low incidence of tuberosity-related complications.

Objective This study aimed to identify PAX9 variants in non-syndromic tooth agenesis families of Chi-na,as well as to analyze the genotype-phenotype of non-syndromic tooth agenesis caused by PAX9 variants,which can provide a basis for the genetic diagnosis of tooth agenesis.Methods We collected the data of 44 patients with non-syn-dromic oligodontia who underwent treatment at Stomatological Hospital of Hebei Medical University between 2018 and 2023.Whole-exome sequencing was performed on the peripheral blood of the proband and its core family members,and the variants were verified by Sanger sequencing.Pathogenicity analysis and function prediction of the variants were per-formed using bioinformatics tools.The correlation between the genotype of PAX9 variant and its corresponding pheno-type was examined by reviewing 55 publications retrieved from PubMed.The studies involved 232 tooth agenesis pa-tients with PAX9 variants.Results A novel PAX9 c.447delG(p.Pro150Argfs*62)and a reported PAX9 c.406C>T(p.Gln136*)were identified in two Chinese families.Through bioinformatics analysis and three-dimensional structural mod-eling,we postulated that the frameshift variant was pathogenic.The outcome was the premature cessation of PAX9 pro-tein,which caused severe structural and functional deficiencies.Summarizing the PAX9 genotype-phenotype relationship revealed that patients carrying the PAX9 variant commonly led to loss of the second molars.Conclusion We identified the novel PAX9 c.447delG(p.Pro150Argfs*62)in a Chinese family of non-syndromic oligodontia,expanding the known variant spectrum of PAX9.The most susceptible tooth position for PAX9 variants of tooth agenesis was the second mo-lars and the deciduous molars during the deciduous dentition.

OBJECTIVE： To study the anti-inflammatory effect and its mechanism of the extract of Dcsmodium microphyllum， so as to provide experiment reference for further study of D. microphyllum. METHODS： Acute inflammatory model was established by xylene，glacial acetic acid and carrageenan. Using dexamethasone as positive control （0.005 g/kg）， inhibitory effects of intragastric different doses of the extract of D. microphyllum （50， 30， 15 g/kg） on xylene-induced ear swelling in normal mice and adrenalectomized mice， glacial acetic acid-induced permeability increasing of abdominal capillaries in normal mice， carrageenan-   induced paw swelling in normal mice and adrenalectomized mice were investigated. The levels of MDA， SOD and NO in the inflammatory tissue of toes of adrenalectomized mice were detected in carrageenan-induced inflammation model. Blank group was set for control （ig. equal volumn of water）. RESULTS： Compared with blank group， ear swelling degree of normal mice and adrenalectomized mice were decreased significantly in D. microphyllum extract high-dose and medium-dose groups while inhibitory rate of ear swelling was increased significantly； the permeability of abdominal capillaries of normal mice was significantly decreased in D. microphyllum extract groups； the swelling degree of toes in normal mice of D. microphyllum extract high-dose and middle-dose groups and adrenalectomized mice were significantly decreased while inhibitory rate of toe swelling was increased significantly （P＜0.05 or P＜0.01）. The levels of MDA and NO in the toe inflammatory site of adrenalectomized mice were decreased significantly in D. microphyllum extract high-dose and medium-dose groups， while the level of SOD was increased significantly （P＜0.05）. CONCLUSIONS： D. microphyllum extract can inhibit acute inflammation in mice significantly. Its anti-inflammatory mechanism is associated with decreasing MDA and NO while increasing SOD levels， and the anti-inflammatory effect does not depend on the hypothalamus-pituitary-adrenal axis system.