Small interfering RNA (siRNA) has a promising future in the treatment of ocular diseases due to its high efficiency, specificity, and low toxicity in inhibiting the expression of target genes and proteins. However, due to the unique anatomical structure of the eye and various barriers, delivering nucleic acids to the retina remains a significant challenge. In this study, we rationally design PACD, an A-B-C type non-viral vector copolymer composed of a hydrophilic PEG block (A), a siRNA binding block (B) and a pH-responsive block (C). PACDs can self-assemble into nanosized polymeric micelles that compact siRNAs into polyplexes through simple mixing. By evaluating its pH-responsive activity, gene silencing efficiency in retinal cells, intraocular distribution, and anti-angiogenesis therapy in a mouse model of hypoxia-induced angiogenesis, we demonstrate the efficiency and safety of PACD in delivering siRNA in the retina. We are surprised to discover that, the PACD/siRNA polyplexes exhibit remarkable intracellular endosomal escape efficiency, excellent gene silencing, and inhibit retinal angiogenesis. Our study provides design guidance for developing efficient nonviral ocular nucleic acid delivery systems.

Objective　To understand the impact of early and timely treatment and initial antiviral treatment regimen on mortality and attrition of antiretroviral therapy. Methods A retrospective cohort study was conducted using download data on antiretroviral therapy for HIV-infected patients in Liuzhou City, Guangxi Province, from the database of the Basic Information System for AIDS Control and Prevention (BISAC) from 2010 to 2020. The Cox proportional risk regression model was used to analyze the influencing factors of mortality and attrition. Results A total of 15 713 infected patients were included, including 53.4% aged 18-<50 years, 69.4% male, 61.0% farmer, 75.1% CD4 count <350 cells /μL before initial antiviral treatment, the overall mortality rate was 4.30/100 person-years, and the overall attrition was 2.42/100 person-years. The results of Cox regression analysis showed that the influencing factors of mortality were pretreatment CD4 counts of 350-<500 cells/μL(AHR=0.72, 95%CI: 0.63-0.81) and ≥500 cells/μL (AHR= 0.64, 95%CI: 0.55-0.76); duration from diagnosis to initial antiviral treatment 91-180 days (AHR=1.25, 95%CI: 1.08-1.45), 181-365 days (AHR=1.26, 95%CI: 1.08-1.47), and ≥365 days (AHR=1.26, 95%CI: 1.11-1.44); initial antiviral treatment regimens of D4T+3TC+EFV/NVP (AHR=1.47, 95%CI: 1.32-1.63) and AZT/D4T/TDF+3TC+LPV/r (AHR=1.73, 95%CI: 1.50-1.99). Factors affecting attrition were pretreatment CD4 counts of 350-499 cells/μL (AHR=1.32, 95%CI: 1.16-1.50) and ≥500 cells/μL (AHR=1.28, 95%CI: 1.10-1.50); interval from HIV positivity confirmation to initial dosing ≥365 days (AHR=1.21, 95%CI: 1.04-1.40), initial antiviral treatment regimens of TDF+3TC+NVP (AHR=1.32, 95%CI: 1.13-1.55), AZT+3TC+EFV/NVP (AHR=1.43, 95%CI: 1.26-1.62) and AZT/D4T/TDF+3TC+LPV/r (AHR=1.33, 95CI%: 1.06-1.67). Conclusions　Early and timely treatment and the initial antiviral treatment regimen of TDF+3TC+EFV have good efficacy, but attention should be paid to the high risk of attrition of HIV-infected people with high CD4 count before treatment.

Objective:To investigate the predictive value of Naples prognostic score (NPS) in assessing the overall survival of intrahepatic cholangiocarcinoma (ICC) patients after receiving hepatectomy treatment.Methods:Clinicopathological characteristics and follow-up data of 164 ICC patients who underwent curative hepatectomy at Zhejiang Cancer Hospital from Jan 2010 to Aug 2022 were retrospectively collected. NPS was calculated basing on preoperative serum albumin concentration, total cholesterol concentration, the neutrophil-lymphocyte ratio, and lymphocyte-monocyte ratio. The relationship between NPS and overall survival was analyzed, and the efficacy of NPS in predicting long-term survival was compared to TNM staging system and other independent risk factors.Results:Multivariate analysis identified the NPS [Score 1 versus 0: 1.864 (1.011-3.437), P=0.046; Score 2 versus 0: 3.013 (1.465-6.199), P=0.003] as an independent risk factor for overall survival. The area under curve (AUC) of the rece中iver operating characteristic (ROC) curve for predicting 5-year OS based on NPS is 0.75, which is higher than TNM staging (0.59) and other independent risk factors (CA19-9:0.71, lymph node metastasis: 0.66, tumor size: 0.62, microvascular invasion: 0.56). Conclusion:NPS as an independent predictor of overall survival for ICC patients, is more accurate than TNM staging system and other clinicopathological factors.

Objective:To investigate the prevalence of pretreatment drug resistance and the genetic polymorphism of CRF08_BC strains among HIV-1 patients in China.Methods:This cross-sectional survey involved the plasma samples of HIV patients in a national pretreatment HIV drug resistance survey conducted in 2018. RNA was extracted from the samples. The fragments containing protease and partial reverse transcriptase (PR/RT) regions were obtained and sequenced. Drug resistance was analyzed using Stanford HIVdb Program. Differences in polymorphic mutations between drug-resistant and non-drug-resistant HIV-1 strains were analyzed by Chi-square test or Fisher′s exact test. The association between drug-resistant and polymorphic mutations was evaluated using CorMut R package. Molecular transmission networks were constructed using HIV-TRACE software. Results:Totally 465 partial pol sequences were obtained from individuals with CRF08_BC infection in 25 provinces and cities. The total pretreatment drug resistance rate was 17.8% (83/465). The pretreatment drug resistance rates to non-nucleoside reverse transcriptase inhibitors (NNRTIs), nucleoside reverse transcriptase inhibitors (NRTIs) and protease inhibitors (PIs) were 16.6% (77/465), 1.1% (5/465) and 0.9% (4/465), respectively. The resistance rate to rilpivirine (RPV) was the highest (15.7%, 73/465). The most common mutation was E138A (11.6%, 54/465). There were six polymorphic mutations (S162C, K102Q, T200A, V179E, I202V, T200M) that co-variated with E138A. The molecular transmission network showed that patients infected with CRF08_BC strains carrying the resistant mutations at position E138 mainly gathered in clusters in Yunnan and Sichuan, and the highest degree of connection was in Lincang, Yunnan. Conclusions:In China, HIV-1 CRF08_BC-infected patients showed a high rate of pretreatment resistance to one of the second-generation NNRTIs, namely RPV. Further researches were warranted to evaluate the impacts of co-mutations of the E138A mutation and polymorphic sites on HIV resistance and replicative capacity.

Objective:To analyze the characteristics of viral isolation and unique recombinant from untreated HIV-1 patients infected through sexual transmission and injection drug use, so as to provide evidence for understanding the biological characteristics and precise prevention and control of HIV-1 infection in different transmission routes.Methods:In view of the different HIV-1 transmission risks, newly diagnosed untreated HIV-1 patients from Beijing, Guangxi and Sichuan were carefully selected. Venous blood was collected to detect the viral load and CD4 + T cell count, and peripheral blood lymphocytes were isolated for virus isolation. Viral RNAs were extracted from the virus supernatant, and the near full-length genome sequences were obtained using in-house method, then the recombination patterns were determined. Results:Among the 65 HIV-1 infection, 32(49.2%), 20(30.8%) and 13(20.0%) were infected via men who have sex with men (MSM), heterosexual and injection drug use (IDU), respectively; genotypes mainly included 26(40.0%) CRF07_ BC, 23(35.4%) CRF01_ AE, and 9(13.8%) unique recombinant types (URFs). A total of 46 HIV-1 clinical strains were isolated. The positive rate of HIV-1 isolation was significantly negatively correlated with CD4 + T cells ( X2=4.22, P=0.04), but positively correlated with viral load ( X2=22.4, P<0.001); the multi-variate generalized estimating equations(GEE) model analysis of HIV-1 P24 antigen content showed similar result. In addition, GEE model showed a positive correlation between viral P24 antigen content and virus-producing culture time (52.14, 95% CI: 9.42~94.87, P=0.017). Viral growth curve analysis showed that the level of viral P24 antigen in MSM Group was significantly higher than that in heterosexual group and IDU group (adjusted P values were p<0.01 and P<0.05, respectively), on the 14th day after culture. The proportion of URFs in MSM Group was higher than that in heterosexual group, and the recombinant breakpoints in MSM Group were more than that in heterosexual group. Conclusions:MSM population was more sensitive to HIV-1 virus isolation; there was unique diversity of recombinant forms of HIV-1 among those with sexually transmitted infections, especially in the MSM population.

Objective To quantitatively judge the degree of tibial bone healing using the finite element wall thickness analysis method, so as to provide an intuitive diagnostic basis for clinical judgment of tibial union and delayed bone healing. Methods After three-dimensional (3D) modeling for the affected and healthy limb side of 48 patients, the maximum wall thickness (MWT) was calculated, and the ratio (B value) was used as a quantitative index of bone healing. When both BMWT2 and BMWT1 were greater than 0.9, bone healing could be judged. When BMWT2 was between 0.9 and 0.7, bone union was judged to be poor, and there was no significant increase in this value after regular reexamination. When BMWT3 was above 0.9 while both BMWT1 and BMWT2 were smaller than 0.7, it could be judged as internal fixation failure, which should be replaced during the second operation. The clinical diagnosis was revised twice, and the final clinical healing results were observed. Results Clinical diagnosis analysis and finite element wall thickness analysis were carried out in 48 patients during each review period, and 21 cases of delayed bone healing and 27 cases of bone nonunion were judged clinically. Among them, 2 cases were judged to be ineffective, and bone grafting intervention was adopted to replace the internal fixation, 12 cases were judged to be still effective, and all cases were finally healed by surgical intervention of bone grafting alone. By Bowker test, P=0.094 was obtained, indicating that the wall thickness analysis method was consistent with the clinical diagnosis. Conclusions The wall thickness analysis method can be used to quantitatively analyze the degree of bone healing at fracture end and realize the rapid calculation of bone healing degree. The case results in this study show that the finite element wall thickness analysis method is superior to the simple clinical diagnosis method, and has better differential diagnostic significance for early diagnosis of poor bone healing.

Objective:To understand influencing factors on the deaths of HIV/AIDS patients receiving antireviral treatment in Butuo county of Liangshan Yi Autonomous Prefecture (Liangshan) from 2010 to 2019, to provide data for drug replacement and sustainable antiviral treatment strategy.Methods:A matched case-control study was used to collect basic and follow-up information on AIDS death patients receiving antiviral treatment in Butuo county of Liangshan from 2010 to 2019. The control group was formed by sampling twice the number of cases. The logistic regression model was used to analyze the risk factors affecting mortality.Results:In 3 355 patients of HIV/AIDS treated with antiviral therapy, 1 179 cases in the death group and 2 176 cases in the control group. Including 81.34% were 30-49 years old, 69.09％males, 99.55% Yi nationality, 91.12% were married or cohabitated, 95.77% had junior high school education or below, and 88.41% peasants. Amultivariate logistic stepwise regression model showed that among the death risk factors, age ≥50 years old was 5.08 times (95% CI:3.05-8.48) that of the 18-29, female was 0.70 times (95% CI: 0.52-0.94) than male, the transmission rate of intravenous drug use was 1.43 times (95% CI: 1.06-1.91) that of heterosexual transmission, CD4 +T lymphocyte (CD4) count ≥350 cells/μl before treatment was 0.38 times (95% CI: 0.30-0.48) that of CD4 <200 cells/μl before treatment, the most recent antiviral treatment regimen containing LPV/r was 0.04 times (95% CI: 0.01-0.18) than that of stavudine (d4T) + lamivudine (3TC) + nevirapine (NVP)/efavirenz (EFV) regimen, drug resistance was 3.40 times (95% CI: 2.13-5.42) of non-drug resistance, non-viral load and non-drug resistance test results were 12.98 times (95% CI: 10.28-16.40) of non-drug resistance. Conclusions:Age, gender, transmission route, CD4 before treatment, the latest antiviral treatment program, and drug resistance test after antiviral therapy were the influencing factors of HIV/AIDS death in Butuo county. It is necessary to expand the coverage of viral load and drug resistance test to change the antiviral therapeutic schedule scientifically and carry out publicity and education on the compliance of patients with antiviral treatment and medical staff training in order to reduce the mortality of patients with antiviral treatment.

Objective:To explore the resistance to protease inhibitors (PI), nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs) and integrase inhibitors (INSTIs) of HIV-1 CRF55_01B and the transmission of drug-resistant strains among HIV-1 CRF55_01B infected patients before antiviral treatment in China.Methods:HIV-1 RNA was extracted from plasma samples of the patients infected with CRF55_01B in the national surveillance of HIV drug resistance before antiviral treatment in 2018. A 1 056 bp gene fragment of protease/reverse transcriptase (PR/RT) region and an 846 bp gene fragment of integrase (IN) region were obtained and sequenced. Drug resistance was analyzed by using all drugs included in the Stanford University HIV db Program, HIV-1 molecular network analysis was performed with software HIV-TRACE and polymorphism mutations of CRF55_01B integrase gene region were analyzed.Results:A total of 178 samples from 26 provinces, municipalities and autonomous regions in China were analyzed, and 170 sequences of CRF55_01B PR/RT region and 170 sequences of IN region of corresponding samples were obtained. The drug resistance rate was 15.3% (26/170). The drug resistance rates of PIs, NRTIs, NNRTIs and INSTIs were 1.2% (2/170), 1.2% (2/170), 15.3% (26/170), 0.6% (1/170), respectively. The level of drug resistance was mostly low. NNRTIs drug resistance mutations were mainly V179D/E co-appeared with other mutations, and 84.1% (143/170) of the infected patients carrying V179D/E alone showed potential drug resistance. INSTIs drug resistance mutation was G163R, and showed low resistance to EVG and RAL. The molecular network access rate was 30.0%(51/170)according to the 0.9% gene distance threshold. The resistant strains were transmitted between men with homosexual transmission and heterosexually transmitted people, and both carried resistant mutations E138G and V179E. In the integrase region, CRF55_01B and CRF01_AE and B subtypes showed high mutation frequency difference in 5 sites (T215A、G134N、I135V, K136R and L101I/V).Conclusions:Before antiviral treatment, CRF55_01B infected patients in China had a high resistance to NNRTIs. Strains carrying both E138G and V179E resistance mutations were transmitting in clusters. The prevalence of CRF55_01B integrase inhibitor resistant strains is low, but some genetic polymorphisms with high mutation rate in the integrase gene region have potential influence on drug sensitivity. The influence of drug resistance of new recombinant strains on antiviral therapy in China needs to be further monitored and analyzed.

Objective:To explore the clinical value of procalcitonin (PCT) in predicting mortality of patients with acute biliary pancreatitis (ABP).Methods:The clinical data of 196 ABP patients admitted in the emergency department of Ruijin Hospital Affiliated to Shanghai Jiaotong University Medical College from January 2013 to June 2017 were analyzed retrospectively. The enrolled patients were divided into survival group ( n=176) and death group ( n=20) according to clinical outcome, and their clinical characteristics, laboratory results(including WBC, CRP, PCT), APACHEⅡ score, BISAP score, modified Marshall score, SOFA score and CTSI at admission were compared between two groups. The ROC curve and AUC were used to evaluate the effectiveness of PCT and multiple scoring systems in predicting mortality in ABP patients, and the Delong test was used to compare the predictive efficacy of various methods at 1-2 d, 3-4 d, and 5-7 d days after onset. Results:The PCT level, APACHEⅡ score, BISAP score, modified Marshall score, SOFA score, and CTSI of patients in the death group were significantly higher than those in the survival group [6.98(3.12, 13.64) μg/L vs 0.55(0.17, 1.74) μg/L, 12.00(6.00, 18.75) vs 6.00(3.00, 9.00), 3.20±1.47 vs 1.59±1.05, 2.85±0.37 vs 1.96±0.64, 5.50(4.00, 9.50) vs 2.00(1.00, 4.25), 5.05±2.33 vs 3.39±1.74], and all the differences were statistically significant (all P values <0.05). The AUC of PCT for predicting death was 0.881 (95% CI 0.820-0.938)and the cut-off value was 2.44. The predictive value of PCT was similar to that of the modified Marshall score, BISAP score and SOFA score, but higher than that of APACHEⅡ score and CTSI (all P values <0.05). The predictive AUC of PCT at 3-4 days after onset was higher than that of modified Marshall score, BISAP score and SOFA score, and were significantly higher than those at 1-2 days after onset. Conclusions:PCT can be used to predict the mortality of ABP within 7 days of onset. The predictive value of PCT was comparable to the modified Marshall score, BISAP score and SOFA score, and the best predictive time was 3-4 days after onset.

Objective:To investigate the consistency of reports of adverse reactions to chemotherapy in patients with stomach neoplasms, main caregivers and nursing records.Methods:By convenient sampling method, a total of 102 pairs of patients who underwent radical gastrectomy and received chemotherapy in the First Affiliated Hospital of Zhengzhou University from June 2018 to June 2019 and their main caregivers were selected. After the second and fourth cycles of chemotherapy, the incidence of adverse reactions to chemotherapy in patients was collected from records patients themselves, main caregivers and nursing staff, and the severity of adverse reactions was assessed using the Chinese version of M.D. Anderson Symptom Inventory (MDASI) . At the same time, Kappa test was used to analyze the consistency of reports of patients, their main caregivers and nursing staff to the 13 adverse reactions of chemotherapy in the scale.Results:After the second chemotherapy cycle, except for the two adverse reactions of fatigue and distress, the severity of other adverse reactions reported by the patients was not statistically significantly different compared with the records of the main caregivers and nursing staff ( P>0.05) . The incidence of fatigue, distress, sleep disturbance, nausea and dry mouth reported by the patients was higher than that recorded by the primary caregivers and nursing staff, and the differences were statistically significant ( P<0.05) . The patients showed good consistency with main caregivers in 10 aspects such as appetite loss, shortness of breath, sleep disturbance, vomiting, pain, nausea, dry mouth, drowsiness, forgetfulness and numbness ( P<0.05) . The patients showed good consistency with the nursing staff in 9 aspects such as appetite loss, shortness of breath, vomiting, pain, nausea, dry mouth, lethargy, forgetfulness and numbness ( P<0.05) . The main caregivers showed good consistency with the nursing staff in 10 aspects such as appetite loss, distress, shortness of breath, vomiting, pain, nausea, dry mouth, lethargy, forgetfulness and numbness ( P<0.05) . After the fourth chemotherapy cycle, the self-reported severities of fatigue, distress, sleep disturbance and sadness of patients were higher than those recorded by the main caregivers and nursing staff, and the differences were statistically significant ( P<0.05) . The incidence of fatigue, distress, sleep disturbance, sadness and dry mouth reported by patients were higher than those recorded by the main caregivers and nursing staff, and the differences were statistically significant ( P<0.05) . The patients showed good consistency with the primary caregivers in 8 aspects such as appetite loss, shortness of breath, vomiting, pain, nausea, dry mouth, forgetfulness and numbness ( P<0.05) , and they showed good consistency with the nursing staff in 7 aspects such as appetite loss, vomiting, pain, nausea, dry mouth, forgetfulness and numbness ( P<0.05) . The primary caregivers showed good consistency with the caregivers in 8 aspects such as appetite loss, sleep disturbance, vomiting, pain, nausea, dry mouth, forgetfulness and numbness ( P<0.05) . Conclusions:Nursing staff and the main caregivers of patients underestimate the incidence and severity of chemotherapy-related adverse reactions in patients with stomach neoplasms. Simple nursing records cannot accurately reflect the patients' chemotherapy symptoms, so self-reports of patients should be considered as one of the reference data.