Objective Cloning-sequencing is a common method to detect the number of trinucleotide repeats.The aim of the present study is to discuss its reliability.Methods One clinically diagnosed SCA1 patient was recruited in the study.The numbers of CAG repeats in ATXN1 gene were estimated via polymerase chain reaction (PCR) and denaturing polyacrylamide gel electrophoresis (DPAGE).To verify accuracy of CAG numbers estimated, the PCR products were electrophoresed on a 2.5% agarose ge] and separated bands were excised for direct sequencing.Also, the longer separated band underwent cloning-sequencing using a TA cloning kit.Results The patient was identified as SCA1 by DPAGE.After direct sequencing, the numbers of CAG repeats were 26 and 47 in the shorter and longer bands, respectively.However, after cloning-sequencing of the longer band, there are 10 different numbers of CAG repeats, including 50, 47, 46, 41,32, 28, 27, 26, 25 and 24.Furthermore, there are other kinds of trinucleotide repeats, such as CCG, CGG, CTG, CAA and TAT scattered among the CAG repeats.Conclusions It is not reliable to identify the number of trinucleotide repeats by cloning-sequencing alone.To improve the reliability, it is better to combine cloning-sequencing with other methods.

The objectives of this study are to prepare resveratrol loaded mixed micelles composed of poloxamer 403 and poloxamer 407, and optimize the formulation in order to achieve higher drug solubility and sustained drug release. Firstly, a thin-film hydration method was utilized to prepare the micelles. By using drug-loading, encapsulation yield and particle size of the micelles as criteria, influence of three variables, namely poloxamer 407 mass fraction, amount of water and feeding of resveratrol, on the quality of the micelles was optimized with a central composite design method. Steady fluorescence measurement was carried out to evaluate the critical micelle concentration of the carriers. Micelle stability upon dilution with simulated gastric fluid and simulated intestinal fluid was investigated. The in vitro release of resveratrol from the mixed micelles was monitored by dialysis method. It was observed that the particle size of the optimized micelle formulation was 24 nm, with drug-loading 11.78%, and encapsulation yield 82.51%. The mixed micelles increased the solubility of resveratrol for about 197 times. Moreover, the mixed micelles had a low critical micelle concentration of 0.05 mg · mL(-1) in water and no apparent changes in particle size and drug content were observed upon micelles dilution, indicating improved kinetic stability. Resveratrol was released from the micelles in a controlled manner for over 20 h, and the release process can be well described by Higuchi equation. Therefore, resveratrol-loaded poloxamer 403/407 mixed micelles could improve the solubility of resveratrol significantly and sustained drug release behavior can be achieved.

Objective To study the protective effect of hydrogen-rich saline (HRS) on apoptosis in skin flap after ischemia/reperfusion injury.Methods Total 18 Sprague-Dawley rats were randomly divided into three groups:a HRS treated group and two physiological saline treated groups (controls 1,2).The rats were anesthetized and an extended abdominal skin flap (6 cm × 9 cm) was elevated in each animal.Ischemia was induced by clamping the left right pedicle for 3 h,then HRS was administered intraperitoneally 10 min before reperfusion,and physiological saline was injected in control groups 1 and 2.In the control group 2,the flaps were elevated without occluding the artery and vein.Five days postoperation,apoptosis,TNF-α level in flap were measured with ELISA,NF-κB in nucleus was determined by Westernblot.Results Apoptotic rate represented (39.72±8.09) ％in HRS group and (69.43±13.27) ％ in control group 1,respectively.Treatment with HRS resulted in a marked reduction in apoptotic rate.TNF-α level was (516.408±38.674) pg/ml in the control group 1,a significant reduced TNF-α was measured in HRS group,accounting for (269.136 ±24.530) pg/ml.Moreover,NF-κB activation was significantly down-regulated by HRS.In control group 2,no significant apoptosis was observed because of non-blood occlusion,and there was no marked elevation of TNF-α and NF-κB.Conclusions HRS can protect skin from ischemia/reperfusion injury,attenuate apoptosis in flaps,which may be associated with the inhibition of TNF-α and NF-kb elevation.

Objective To investigate the correlation between survival motor neuron (SMN) gene deletion and Chinese patients with sporadic amyotrophic lateral sclerosis (SALS).Methods A total of 141SALS patients and 134 unrelated controls were recruited from the Chinese population.Polymerase chain reaction (PCR) and restriction fragment length polymorphisro (RFLP) analysis were performed to screen SMN gene deletion.Frequencies of deletion were coropared by Chi-square test.Results Four patients and 3 controls were detected to have horoozygous SMN2 deletion.The frequencies of SMN2 deletion were 2.84%(4/141) and 2.24% (3/134), respectively, which was not significantly different (χ2= 0.0001, P =1.000).No subjects were found to have homozygous SMN1 deletion.Condusion There is no correlation between SMN gene deletion and Chinese patients with SALS.

Using high pressure homogenization method combined with spray-drying, budesonide-loaded chitosan microparticles were prepared and the in vitro release profile was investigated. The microparticles were then blended with lactose using a vortex mixer, influence of mixing speed, mixing time on drug recovery rate and content homogeneity were investigated. Meanwhile, influence of lactose content on drug recovery rate, content homogeneity, powder flowability and in vitro deposition were studied. It turned out that budesonide was released from the microparicles in a sustained manner, with fine particle fraction as high as 46.0%, but the powder flowability was poor. After blending with 10 times of lactose, the drug recovery rate was 96.5%, with relative standard deviation of drug content 2.5%, and fine particle fraction of the formulation increased to 59.6% with good flowability. It's demonstrated that using a vortex mixer, budesonide sustained-release dry powder for inhalation with good recovery and content homogeneity could be prepared, the formulation had good flowability and was suitable for pulmonary inhaling.

Objective To investigate the correlation between setup error and couch position error in radiotherapy.Methods A total of 25 patients with thoracic and abdominal tumors who recently finished image-guided radiotherapy were randomly selected.The data on couch position during treatment were obtained through the record validation system, and then the couch position error was calculated.The Pearson correlation analysis was used to investigate the correlation between setup error and couch position error during treatment.Results In the ≥5 setup errors among the 25 patients, the correlation coefficient between random setup error and random couch position error was 0.83(P=0.00), and the correlation coefficient between systematic setup error and systematic couch position error was 0.36(P=0.11).Conclusions In radiotherapy, the random setup error is highly correlated with the random couch position error, while a moderate or low correlation exists between the systematic setup error and the systematic couch position error.

Objective:To evaluate the application of visual feedback coaching method, which is embedded in an optical surface monitoring system, in deep inspiration breath holding during the radiotherapy in left breast cancer patients after breast-conserving surgery.Methods:Thirty patients with left breast cancer, who were scheduled to receive the whole breast radiotherapy after breast-conserving surgery, met the requirements of deep inspiration breath holding after respiratory coaching with the visual feedback coaching module in the optical surface monitoring system. Active breathing control equipment was used to control breath-holding state and CT simulation was performed. During treatment, optical surface monitoring system was used to guide radiotherapy. All patients were randomly divided into two groups. In group A ( n=15), visual feedback respiratory training method was utilized and not employed in group B ( n=15). In group A, the visual feedback coaching bar of the optical surface monitoring system was implemented, while audio interactive method was employed to guide patients to hold their breath. Real-time data of optical body surface monitoring were used to compare the interfraction reproducibility and intrafraction stability of breath holding fraction between two groups. Besides, the number of breath holding and treatment time per fraction were also compared. GraphPad prism 6.0 software was used for data processing and mapping, and SPSS 21.0 software was used for analyzing mean value and normality testing. Results:Compared with the control group, the reproducibility in the experiment group was reduced from 1.5 mm to 0.7 mm, the stability was reduced from 1.1 mm to 0.8 mm, the mean number of breath holding required per fraction was decreased from 4.6 to 2.4, the mean beam-on time per fraction from 336 s to 235 s, and the treatment time per fraction was shortened from 847 s to 602 s (all P<0.05), respectively. Conclusions:The application of visual feedback coaching method can improve the reproducibility and stability of breath holding during radiotherapy for left breast cancer, and it can also effectively reduce the number of breath holding and shorten the treatment time per fraction.

Objective To explore the categories of dyadic mental health literacy among stroke patients and their caregivers,and to analyze the differences in the characteristics of different classes of stroke patients and their caregivers.Methods A convenient sampling method was used to select 287 dyads of stroke patients and their caregivers who were treated at a tertiary general hospital in Henan province from July to October 2020.The general information questionnaire,Multicomponent Mental Health Literacy and Social Support Rating Scale were used for investigation.Latent profile analysis was adopted to explore the categories of dyadic mental health literacy,and multiple logistic regression was used to analyze the influencing factors of each category.Results Stroke patients and their caregivers were divided into 4 categories based on scores of mental health literacy:dyadic low resource group(19.86％),dyadic low literacy group(54.36％),dyadic low belief group(11.15％),dyadic high literacy group(14.63％).The patient's age,average monthly household income,the caregiver's age,the caregiver's Social Support Rating Scale score,the caregiver's educational level,daily care time and total time length of care were the factors influencing the categories of dyadic mental health literacy among stroke patients and their caregivers(P＜0.05).Conclusion Stroke patients and their caregivers were divided into 4 categories based on scores of mental health literacy.Medical staff should carry out comprehensive psychological interventions for stroke patients and their caregivers with different dyadic mental health literacy characteristics,so as to improve dyadic mental health literacy.

Background::Few evidence is available in the early prediction models of behavioral and psychological symptoms of dementia (BPSD) in Alzheimer’s disease (AD). This study aimed to develop and validate a novel genetic-clinical-radiological nomogram for evaluating BPSD in patients with AD and explore its underlying nutritional mechanism.Methods::This retrospective study included 165 patients with AD from the Chinese Imaging, Biomarkers, and Lifestyle (CIBL) cohort between June 1, 2021, and March 31, 2022. Data on demographics, neuropsychological assessments, single-nucleotide polymorphisms of AD risk genes, and regional brain volumes were collected. A multivariate logistic regression model identified BPSD-associated factors, for subsequently constructing a diagnostic nomogram. This nomogram was internally validated through 1000-bootstrap resampling and externally validated using a time-series split based on the CIBL cohort data between June 1, 2022, and February 1, 2023. Area under receiver operating characteristic (ROC) curves, calibration curves, and decision curve analysis (DCA) were used to assess the discrimination, calibration, and clinical applicability of the nomogram.Results::Factors independently associated with BPSD were: CETP rs1800775 (odds ratio [OR] = 4.137, 95% confidence interval [CI]: 1.276-13.415, P = 0.018), decreased Mini Nutritional Assessment score (OR = 0.187, 95% CI: 0.086-0.405, P <0.001), increased caregiver burden inventory score (OR = 8.993, 95% CI: 3.830-21.119, P <0.001), and decreased brain stem volume (OR = 0.006, 95% CI: 0.001-0.191, P = 0.004). These variables were incorporated into the nomogram. The area under the ROC curve was 0.925 (95% CI: 0.884-0.967, P <0.001) in the internal validation and 0.791 (95% CI: 0.686-0.895, P <0.001) in the external validation. The calibration plots showed favorable consistency between the prediction of nomogram and actual observations, and the DCA showed that the model was clinically useful in both validations. Conclusion::A novel nomogram was established and validated based on lipid metabolism-related genes, nutritional status, and brain stem volumes, which may allow patients with AD to benefit from early triage and more intensive monitoring of BPSD.Registration::Chictr.org.cn, ChiCTR2100049131.

Objective To evaluate the effect of setup errors upon the target area and the organs at risk (OAR) during radiotherapy for prostate cancer.Methods Twelve prostate cancer patients receiving treatment in the recent 1 year were randomly recruited in this study.The position of each patient was verified by using cone beam CT (CBCT) for 6-10 times during the treatment.In treatment planning system (TPS),the isocenter position was moved along the setup errors with averaging error value (Plan_A) and each CBCT value (Plan_F).The dose distribution was recalculated without changing the beam setting,weight factors and monitor units (MUs).The dose difference was statistically compared between the simulation and original plans (Plan_O).Results For clinical target volume (CTV) D95,there was a significant difference between Plan_A and Plan_O (P =0.008),whereas no significant difference was observed between Plan_F and Plan_O.There were significant differences between Plan_F and Plan_O,Plan_A and Plan_O (P=0.004,and 0.041) for the planned target volume (PTV) D95.For OAR,rectal V60,Dmax,left femoral V20,Dmax and right femoral Dmax significantly differed between Plan_F and Plan_O (P=0.026,0.015,0.041,0.049,0.003).However,only left femoral Dmax significantly differed between Plan_A and Plan_O (P=0.045).The movement in the superior-inferior (SI) direction was significantly correlated with the changes in the rectal V40,V50 and V60 and PTV D95 (r=-0.785,-0.887,-0.833,0.682).The movement in the anterior-posterior (AP) direction was significantly associated with the variations in the bladder V20,V30,V40,V50 and V60(r=-0.945,-0.823,-0.853,-0.818,-0.774).The evaluation indexes of all normal tissues in the re-plan could meet the clinical requirements.However,the volume of target prescription volume had different levels of deficit,and the deficit of Plan_F was greater than that of Plan_A.Conclusions The simulation results of averaging into the TPS underestimates the effect of daily setup errors on the dose distribution.The effect of setup errors on the dose distribution in target area is greater than that of normal tissues.Y-direction errors are more likely to cause the variations of the rectal and PTV dose,and the errors in the z-direction are inclined to cause the changes in the bladder dose.