ObjectiveTo establish a vaginal self-sampling HPV cervical cancer screening model in Hainan Province, to analyze the application of p16 protein detection in HPV positive and non-HPV16 /18 shunt screening. MethodsFrom January 2019 to September 2022, a total of 200 women from the targeted population was randomly selected for vaginal self-sampling HPV typing test to screen cervical cancer using randomized numeric table method, followed by cervical cytology sampling for cytology p16 protein detection. Postoperative pathological examination was used as the gold standard. Multivariate logistic regression analysis was used to analyze the influencing factors of HPV positive detection rate in cervical lesions, and the nomogram model was constructed simultaneously. The receiver operating characteristic(ROC) curve and calibration curve were used for evaluating the accuracy of the nomogram model. Differences in the distribution of self-sampled HPV-positive and HPV infected genotypes were recorded, and the application of p16 protein detection in HPV-positive and non-HPV16/18 shunt screening was analyzed. ResultsAged ≥40 years, BMI ≥28.00 kg·m^-2, number of sexual partners ≥2, frequency of sexual life ≥10 times·month^-1, bleeding from sexual intercourse, and age of first sexual intercourse <22 years were the risk factors for HPV positive of cervical lesions (all P<0.001). The results of ROC curve and calibration curve showed that the area under ROC curve (AUC) was 0.874 (95%CI: 0.823‒0.907, P<0.05), the sensitivity was 0.835, the specificity was 0.847, and the Youden index was 0.672, indicating a good fit of the model. Results of vaginal self-sampling HPV test showed that the positive rate of HPV was 86.50% (173/200). HPV high-risk infection types mainly included HPV16, 18, 31, 33, 35, 39, 45, 52, 56, 58, 59, 68, 73, and 82. Single HPV infection accounted for 95.95% (166/173), 2.89% (5/173) were infected with two types of HPV, and 1.16% (2/173) were infected with three or more types of HPV. Colposcopic pathologic diagnosis was used as the gold standard, and the results showed that the accuracy of p16 protein detection in the diagnosis of cervical cancer was 93.50% (187/200), with a sensitivity of 96.53% (167/173), and a specificity of 74.07% (20/27). The negative and positive predictive value were 76.92% (20/26) and 95.98% (167/174), respectively. The results of shunt screening showed that there were 80 cases infected with HPV16, 79 cases infected with HPV18 and 41 cases of non-HPV16/18, with a sensitivity of 90.91%, 90.32% and 86.67%, a specificity of 71.43%, 64.71% and 72.73%, a negative predictive value of 62.50%, 64.71% and 66.67%, a positive predictive value of 93.75%, 90.32% and 89.66%, and an accuracy of 87.50%, 84.81% and 82.93%, respectively. The specificity and accuracy of p16 positive screening for cervical cancer were significantly higher than that of HPV positive detection, but the false positive rate was significantly lower than that of HPV positive detection. The AUCs of HPV positive, p16 positive and combination of the two detection methods for cervical cancer were 0.603, 0.822 and 0.907, respectively. ConclusionVaginal self-sampling HPV testing is a widely accepted mode for cervical cancer screening. Cervical cytology p16 protein detection is important for self-sampled HPV positive and shunt screening of non-HPV16/18.

Objective To explore the clinical efficacy and safety of donepezil(DNPQ)combined with butylphthalide sequential therapy(BST)in the treatment of Parkinson's syndrome(PS).Methods In this study,104 patients with Parkinson's disease(PD)who were diagnosed and treated in the Department of Neurology of The Fourth Hospital of Changsha from January 2020 to November 2023 were randomly divided into a control group(butylphthalide softcapsule combined with DNPQ)and an observation group(BST combined with DNPQ).The main observation indicators of this study were the clinical efficacy and drug-related adverse reactions after 3 months of treatment.The secondary observation indicators were the cognitive function[Montreal Cognitive Assessment(MoCA)and Mini-Mental State Examination(MMSE)],overall condition[Unified Parkinson's Disease Rating Scale(UPDRS)],activity of daily living(ADL),and oxidative stress-related cytokines[recombinant human Parkinson's disease protein 7(PARK7),neurotrophic factor 3(NT3),and C-reactive protein(CRP)]improvement after treatment.Results There were 52 patients in the experimental group and 52 patients in the control group.The treatment efficacy rate in the experimental group was significantly higher than that in the control group(P<0.05),while the incidence of adverse reactions was significantly lower than that in the control group(P<0.05).Before treatment,there were no significant differences in MoCA scores,MMSE scores,UPDRS scores,ADL scores,serum NT3,CRP,and PARK7 levels between the two groups(P>0.05).After treatment,the MoCA score,MMSE score,and ADL score in the experimental group were higher than those in the control group(P<0.05),while the UPDRS score was lower than that in the control group(P<0.05).After treatment,the serum NT3 level in the experimental group was higher than that in the control group(P<0.05),while the serum CRP and PARK7 levels were lower than those in the control group(P<0.05).Conclusion The combination of DNPQ and BST has better clinical efficacy and safety,which can improve cognitive function,ADL and oxidative stress-related cytokine content in patients with PS.

Radiation-induced brain injury (RBI) is the most serious complication of head and neck tumor after radiotherapy. The pathogenesis of RBI is complicated, and the clinical course is irreversible, while no effective treatment available. The activation of glial cells is one of the main theories of RBI, and the prevention and treatment of RBI by targeting glial cells is the focus of current research. As a post-transcriptional regulatory factor, microRNA (miRNA) has been confirmed to be involved in regulatingglial cell radiosensitivity, inflammation type transformation, autophagy, exosomatic, long non-coding RNA (lncRNA), circular RNA (circRNA) and other related pathways, thereby mediating the occurrence and development of cascade reaction of inflammatory injury and neurological function repair of central nervous system (CNS) disease. Therefore, the establishment of miRNA - glial regulatory network may provide a new strategy for the prevention and treatment of RBI.

Objective:To fabricate biogenic gas vesicles (GVs)- polyethyleneimine (PEI)-marrow mesenchymal stem cells (BMSCs) and evaluate its potential on stem cell tracking with ultrasound imaging.Methods:GVs were cationized by PEI to fabricate GVs-PEI. The diameter and zeta potential of GVs-PEI were determined. GVs-PEI were co-incubated with BMSCs to obtain GVs-PEI-BMSCs and stem cell uptake was observed by fluorescence microscope. The cell viability of GVs-PEI-BMSCs was verified by cell counting kit-8 (CCK-8) assay were set. Ultrasound imaging was performed on 0, 2, 4 and 6 d in agarose phantom to evaluate ultrasound imaging capability of GVs-PEI-BMSCs group and BMSCs group in vitro. GVs-PEI-BMSCs and BMSCs were injected into quadriceps femoris of SD rats, and ultrasound imaging was performed on 0, 2, 4 and 6 d to evaluate the ultrasound imaging capability in vivo. One-way analysis of variance and independent-sample t test were used to analyze the data. Results:The diameter and zeta potential of GVs-PEI were (383.63±11.55) nm and (18.48±2.20) mV. Plenty of GVs-PEI were observed in GVs-PEI-BMSCs through microscope. When BMSCs were incubated with GVs-PEI in absorbance ( A) 500 nm of 0.5 and 1.0, there were no significant changes in the cell viability of GVs-PEI-BMSCs at 24, 48 and 72 h ( F values: 7.078-11.982, all P>0.05). Compared with BMSCs, GVs-PEI-BMSCs showed better ultrasound imaging capability in vitro in all time points with still significantly different signal at 6 d (634.29±10.78 vs 2 864.51±100.86; t=-121.86, P<0.001). The ultrasound imaging capability of GVs-PEI-BMSCs in vivo was much better than that of BMSCs at each time point with still significantly different signal at 6 d (2 108.02±217.96 vs 267.71±7.87; t=-121.39, P<0.001). Conclusion:GVs-PEI-BMSCs are successfully fabricated with the advantages of significant ultrasound imaging capability, long duration and safety, which provide a brand-new means for stem cells tracking in vivo.

BACKGROUND: In the treatment of osteoporotic vertebral compression fractures with percutaneous vertebroplasty, the efficacy and safety of bone cement injection by unipedicular and bipedicular approaches are still controversial. Some studies suggest that bone cement injection via unipedicular approach can shorten operation time and reduce postoperative complications, while the other studies suggest that bone cement injection via bipedicular approach can make bone cement distribute more evenly in the vertebral body and relieve pain better. OBJECTIVE: To systematically assess the efficacy and safety of percutaneous vertebroplasty via unipedicular versus bipedicular approach in the treatment of osteoporotic vertebral compression fractures. METHODS: Randomized controlled trials about unipedicular versus bipedicular percutaneous vertebroplasty in the treatment of osteoporotic vertebral compression fracture published before September 18 t h, 2018 were retrieved in the PubMed, Cochrane library, Embase, CNKI, VIP, WanFang data and CBM. Two researchers independently screened all the literatures, carried out data extraction and used improved Jadad to evaluate the methodological quality of the included studies. Meta-analysis using Revam 5.3 was conducted. Egger's test was utilized to evaluate the publication bias. RESULTS AND CONCLUSION: Totally 14 randomized controlled trials including 900 cases were eventually included, 452 cases in unipedicular approach group and 448 cases in bipedicular approach group. The Meta-analysis results showed that compared with the bipedicular approach, the unipedicular approach required shorter operation time [weighted mean difference (WMD) =-16.59, 95％ confidence interval (CI) (-19.25, -13.94), P < 0.001], smaller amount of bone cement injected [WMD=-1.27, 95％ CI (-1.64, -0.89), P < 0.001], and had lower incidence of cement leakage [relative risk=0.70, 95％ CI (0.53, 0.92), P =0.01]. There were no significant differences in short-and long-term Visual Analogue Scale scores, short-and long-term Oswestry Disability Index scores, and the postoperative incidence of adjacent vertebral fractures between the two groups (P> 0.05). Overall, bone cement injection both via bipedicular and unipedicular approaches can lead to a significant improvement in pain relief and living quality of osteoporotic vertebral compression fracture patients, but bone cement injection via unipedicular approach can shorten operation time, reduce cement volume and lower the incidence of cement leakage compared with the bipedicular approach.

Objective To explore the relationship between clinical manifestations and pathological changes in diabetic nephropathy (DN) and to assess the predictive power of the pathologic classification for DN established by the Renal Pathology Society in 2010.Methods Patients with type 1 or type 2 diabetes and biopsy-proven DN in the Third Affiliated Hospital of Sun Yat-sen University between January 2004 to June 2014 were enrolled in the present study and were followed-up until 31 December 2014.The outcome was defined as renal end-points including renal replacement therapy and doubling of serum creatinine as well as all-cause mortality.The laboratory and histologic data were analyzed and outcomes were assessed using survival analysis.Results Fifty-seven people enrolled in this study were categorized into Class Ⅱa (n=9),Class Ⅱb (n=9),Class Ⅲ (n=25) and Class Ⅳ (n=14) while no participants belonged to Class Ⅰ.The changes of Class Ⅱa were slight and those of Class Ⅳ were severe both in the clinical data (diabetic duration,blood pressure,estimated glomerular filtration rate,urine protein excretion rate,albumin and hemoglobin) and the pathological data (percentage of global glomerulosclerosis,percentage and scoring of interstitial fibrosis and tubular atrophy,scoring of interstitial inflammation and incidence of large vessel lesions).There were no significant differences between Class Ⅱb and Ⅲ in the above variables except for the scoring of arteriosclerosis.The mean follow-up duration was 25.9 months.Twenty-five patients (43.9％) reached the renal outcomes and six people (10.5％) reached all-cause mortality.The survival analysis showed that there were significant differences among the renal survival curves of different glomerular classes and of different interstitial and vascular scorings,but not in the survival curves related to all-cause mortality.Conclusions The glomerular classes are not completely associated with renal prognosis.The clinical manifestations and renal outcomes are benign in Class Ⅱa,moderate but similar in Class Ⅱb and Ⅲ and severe in Class Ⅳ.The glomerular classification and interstitial and vascular scorings are associated to renal prognosis while their associations with mortality remain to be verified.

Objective To fabricate iRGD targeted liposome-microbubble complex containing uPA ( iRGD-LMC) ,and to improve the thrombolytic efficiency and reduce the risk of thrombolysis by iRGD-LMC combining with ultrasound targeted microbubbles destruction ( UTMD ) to release drug into the thrombus site with the help of microbubble cavitation effect . Methods Biotinylated iRGD-MBs were fabricated by thin-film rehydration method .Biotinylated liposomes containing uPA were fabricated by freeze-thaw method and were conjugated to the biotinylated iRGD-MBs surface through biotin-avidin linkage . The iRGD-LMC was subjected to confocal microscopy to determine the particle morphology . The concentration , average diameter and size distribution were determined by particle sizing instrument . The uPA loading efficiency was measured by BCA Protein Assay Kit . Ultrasound imaging was performed using a Vevo 2100 ultrasound imaging system . The iRGD-LMC was irradiated by different ultrasound time and intensity to release drug . Thrombolytic effect in vitro of iRGD-LMC combined with UTMD was observed on the thrombosis model which was extracted from mouse blood . Results iRGD-LMC was successfully prepared . iRGD-LMC was exhibited a well-defined spherical morphology and homogeneous distribution ,like ordinary microbubbles . The concentration of iRGD-LMC was ( 0 .51 ± 0 .03 ) × 109 / ml and average diameter was ( 2 .62 ± 0 .12) μm . Drugs loading efficiency was ( 3878 .5 ± 97 .8) μg uPA per 108 microbubbles . iRGD-LMC could achieve contrast-enhanced ultrasound imaging in vitro . The thrombolytic effect of iRGD-LMC +US group ( 87 .66 ± 1 .69) ％ was the best in vitro ,and had significant difference with others groups ( P <0 .05) ,followed by iRGD-LMC group ( 53 .32 ± 4 .86) ％ and uPA group ( 51 .09 ± 9 .01) ％ ,Compared with PBS group ,US group ( 23 .56 ± 9 .46) ％ had thrombolytic effect . Conclusions iRGD-LMC is successfully prepared ,which has the advantages of high drug loading of liposomes and good acoustic properties of microbubbles . iRGD-LMC combined with UTMD achieves a significant thrombolytic effect in vitro .

Objectives The aim of the study was to detect the frequency of B regulatory cells (Breg)and the correlation between Breg and T regulatory cells (Treg) in pancreatic cancer patients,and to investigate its role in pathogenesis of pancreatic cancer.Methods 50 pancreatic cancer patients and 21 healthy controls were enrolled.CD19+CD24hiCD38hi Breg and CD4+CD25high Treg was also determined via flow cytometry.The correlation between Breg and Treg was analyzed by Spearman correlation test.Results Upregulation of CD19+CD24hiCD381hi Breg was associated with pancreatic cancer progression.Furthermore,this B cell subset was positively correlated with the frequency of CD4 + CD25high Treg cells.Conclusions Together,CD19 + CD24hiCD38hi Breg cells are significantly elevated in pancreatic cancer patients,indicating this B cell subset might play an vital role in clinical progression of pancreatic cancer.The significant positive correlation between Breg and Treg may suggest CD 1TCD24hiCD38hi Breg are affecting tumor progression through Treg cells.

Objective To develop a vulnerable plaque targeting ultrasound contrast agent (UCA) and to evaluate its affinity and imaging performance in vitro.Methods E-selectin receptor-targeting UCA,which conjugated with monoclonal antibody of E-selectin,was prepared with filming-rehydration method and biotin-avidin linkage.The size and distribution of UCA were measured with particle size analyzer,the connectivity condition of microbubbles with E-selectin antibody was also detected with fluorescence analysis.The cytotoxicity from microbubble and ultrasound irradiation was evaluated through cell counting kit-8 (CCK8) assay.The adhesion effect of UCA was assessed after co-incubated with activated mouse endothelial cells (bEnd.3) and compared with that of free antibody intervention group and control group.The imaging performance of UCA at different time points was observed on an ultrasound equipment with a high-frequency transducer.Two-sample t test and one-way analysis of variance were performed to analyze the data.Results E-selectin receptor-targeting UCA was successfully prepared.The cytotoxicity result with CCK8 assay demonstrated the favorable biocompatibility of UCA.The connection amount of UCA on activated bEnd.3 cells ((6.23 ± 0.45) bubbles/cell) was significantly higher than that of the free antibody intervention group ((1.57±0.34) bubbles/cell) and control group ((0.07±0.03) bubbles/cell;F=291.43,P＜0.01).The performance of in vitro ultrasonography at the same time points showed no obvious difference between targeting UCA and control UCA (all t＜0.51,all P＞0.05).Conclusions The prepared E-selectin receptor-targeting UCA has favorable targeting and imaging capabilities.It might be a potentially ultrasound molecular imaging agent for early detection and prognosis evaluation of vulnerable plaque.

Wrist-ankle acupuncture is a type of special acupuncture therapy, featured by simple operation, swift efficiency, and few adverse effects. In the recent years, ankle-wrist acupuncture has become more and more popularized in clinic. By retrieving the relevant medical literatures from VIP, CNKI, and Wan Fang databases, this manuscript was to analyze the history of wrist-ankle acupuncture and to review it from both mechanism and clinical reports. Meanwhile, this article also pointed out the existing problems in the research of wrist-ankle acupuncture during the recent years, and expected further standardization and studies.