Latent human immunodeficiency virus（HIV）infection remains the principal barrier to functional cure. Proviruses integrate into multiple immune cell types and persist long term，creating a heterogeneous reservoir. Accurate identification of latent infection and assessment of proviral intactness and function are prerequisites for progress toward cure. Here we compare key reservoir assays and their trade-offs，including HIV DNA quantification by quantitative PCR（qPCR）and droplet digital PCR（ddPCR），detection of replication-competent virus by the quantitative viral outgrowth assay（QVOA），and inducible transcription/protein readouts such as Tat/Rev induced limiting dilution assay（TILDA）and viral protein spot assay（VIP-SPOT）. We also outline applications of spatial transcriptomics and in situ hybridization（RNAscope/DNAscope）for anatomic localization and functional analysis. We also summarize advances in experimental models，spanning in-vitro systems（primary T-cell and myeloid latency models）and in-vivo platforms（humanized mice and rhesus macaque SHIV models）used for mechanism studies and intervention testing. Looking ahead，coordinated use of these orthogonal tools can improve estimates of reservoir size，inducibility，and tissue distribution，and provide a practical platform for translational studies toward functional cure.

Latent human immunodeficiency virus（HIV）infection remains the principal barrier to functional cure. Proviruses integrate into multiple immune cell types and persist long term，creating a heterogeneous reservoir. Accurate identification of latent infection and assessment of proviral intactness and function are prerequisites for progress toward cure. Here we compare key reservoir assays and their trade-offs，including HIV DNA quantification by quantitative PCR（qPCR）and droplet digital PCR（ddPCR），detection of replication-competent virus by the quantitative viral outgrowth assay（QVOA），and inducible transcription/protein readouts such as Tat/Rev induced limiting dilution assay（TILDA）and viral protein spot assay（VIP-SPOT）. We also outline applications of spatial transcriptomics and in situ hybridization（RNAscope/DNAscope）for anatomic localization and functional analysis. We also summarize advances in experimental models，spanning in-vitro systems（primary T-cell and myeloid latency models）and in-vivo platforms（humanized mice and rhesus macaque SHIV models）used for mechanism studies and intervention testing. Looking ahead，coordinated use of these orthogonal tools can improve estimates of reservoir size，inducibility，and tissue distribution，and provide a practical platform for translational studies toward functional cure.

Objective:To learn about the clinical manifestations, laboratory characteristics and treatment of patients with acute and chronic brucellosis.Methods:Clinical data of 153 brucellosis patients admitted to the Guangzhou Eighth People's Hospital, Guangzhou Medical University from 2012 to 2022 were retrospectively collected, including general information, epidemiological characteristics, clinical manifestations, laboratory test results, imaging examination results, treatment and prognosis. According to the course of disease < 180 d and ≥180 d, these patients were divided into acute brucellosis group and chronic brucellosis group, and the clinical data of the two groups of patients were compared and analyzed.Results:A total of 153 patients with brucellosis were included, including 119 in the acute brucellosis group and 34 in the chronic brucellosis group. The age was (46.2 ± 13.8) years old, with 115 males (75.2%) and 38 females (24.8%), and 85 patients (55.6%) were occupational exposed. Complications occurred in 90 patients (58.8%), and the incidence of complications in the acute brucellosis group was lower than that in the chronic brucellosis group [76.5% (26/34) vs 53.8% (64/119), χ 2 = 5.62, P = 0.018]. The most common clinical manifestations were fever and arthralgia, with 128 cases (83.7%) and 124 cases (81.0%), respectively. The incidence of fever in the acute brucellosis group was higher than that in the chronic brucellosis group [87.4% (104/119) vs 70.6% (24/34), χ 2 = 5.46, P = 0.019], while the incidence of arthralgia was lower than that in the chronic brucellosis group [77.3% (92/119) vs 94.1% (32/34), χ 2 = 4.83, P = 0.027]. In laboratory tests, the positive rate of blood culture was 59.5% (91/153), and it was higher in the acute brucellosis group than that in the chronic brucellosis group [67.2% (80/119) vs 32.4% (11/34), P < 0.05]. The incidence of elevated procalcitonin [PCT, 58.6% (58/99) vs 24.1% (7/29), χ 2 = 10.65, P = 0.001] and the incidence of liver dysfunction [33.9% (40/118) vs 15.2% (5/33), χ 2 = 4.33, P = 0.037] in the acute brucellosis group were higher than those in the chronic brucellosis group. In the imaging examination, 61 patients (39.9%) experienced bone destruction, and the incidence of bone destruction in the chronic brucellosis group was higher than that in the acute brucellosis group [55.9% (19/34) vs 35.3% (42/119), χ 2 = 4.68, P = 0.031]. All patients were treated with antibiotics, with a median of 3 and 4 types of antibiotics used in the acute and chronic brucellosis groups, respectively. The overall incidence of adverse drug reactions was 5.2% (8/153). After treatment, 65 cases (42.5%) recovered, 70 cases (45.8%) improved, and 18 cases (11.8%) did not recover. Conclusions:The main clinical manifestations of brucellosis patients are fever and arthralgia, with a high incidence of complications. All patients are treated with combined antibiotics therapy. Patients in acute brucellosis group have a higher incidence of fever, positive blood culture, elevated PCT and abnormal liver function, while patients in chronic brucellosis group have a higher incidence of complications, arthralgia and bone destruction.

Objective:To investigate the efficacy and safety of high-precision transcranial direct current stimulation (tDCS) targeting the anterior prefrontal cortex (aPFC) in prospective memory (PM) deficits in patients with schizophrenia.Methods:A total of 38 schizophrenia patients with PM deficits admitted to Outpatient Department of Psychiatry, Beijing Anding Hospital Affiliated to Capital Medical University from March 2022 to March 2023 were included and divided into true stimulus group ( n=19) and pseudo-stimulus group ( n=19) by random envelope method. Two mA stimulation current intensity with duration of 20 min was given to the true stimulus group, and same stimulation current intensity with duration of 40 s was given to the pseudo-stimulus group twice daily for 5 d. PM function was assessed by Cued Unfocused Laboratory Prospective Memory Task before and 1 week after stimulation, cognitive function and severity of clinical symptoms were evaluated by Positive and Negative Symptom Scale (PANSS) and Chinese version of MATRICS consensus cognition test (MCCB). Safety was assessed by tDCS adverse reaction questionnaire at the end of stimulation. Results:The time (before and 1 week after stimulation) and group interactions of PM trial accuracy and PM trial response time between the two groups were not significantly different ( P>0.05). Compared with that before stimulation, the PM trial accuracy 1 week after stimulation was significantly improved in the true stimulus group ([0.38±0.22] % vs. [0.57±0.28] %, P<0.05). No significant difference in PM trial accuracy ([0.56±0.25] % vs. [0.67±0.25] %) or PM trial response time ([2 216.46±570.03] ms vs. [2 059.59±378.41] ms) between before and 1 week after stimulation was noted in the pseudo-stimulus group ( P>0.05). In terms of severity of clinical symptoms and cognitive function, no significant difference in PANSS or MCCB scores were noted between the true stimulus group and pseudo-stimulus group 1 week after treatment ( P>0.05); no significant difference was noted between the two groups in time (before and 1 week after stimulation) and group interaction of all indexes ( P>0.05). In terms of adverse reactions, compared with the pseudo-stimulus group, the true stimulus group had significantly higher score of "skin redness" ( P<0.05); no significant differences in scores of other adverse reactions were noted between the two groups ( P>0.05). No serious adverse events occurred in all patients. Conclusion:In this study, no positive results have been found in improving PM deficits in patients with schizophrenia with high-precision tDCS targeting aPFC, but existing results suggest an improved trend, which can provide preliminary evidence for subsequent large-sample clinical trials to improve PM deficits in schizophrenia.

In order to establish a stable in vitro culture platform for chicken small intestine three-dimensional (3D) organoids, in this study, crypt cells were collected from the small intestine of 18-day-old embryos of AA broilers. On the basis of the L-WRN conditioned medium, we optimized the culture conditions of chicken small intestinal organoids by adjusting the proportions of nicotinamide, N-acetylcysteine, LY2157299, CHIR99021, Jagged-1, FGF, and other cytokines to select the medium suitable for the long-term stable growth of the organoids. The optimization results showed that the addition of 1.5 µmol/L CHIR99021 significantly improved the organoid formation efficiency and organoid diameter. When 0.5 µmol/L Jagged-1 was added, a small amount of bud-like tissue appeared in organoids. After the addition of 50 ng/mL FGF-2, the rate of organoid germination was significantly increased. The 1.5 µmol/L CHIR99021, 0.5 µmol/L Jagged-1, and 50 ng/mL FGF-2 added in the medium can cooperate with each other to improve the formation and speed up the proliferation and differentiation of organoids, while improving the stemness maintenance of cells. The morphology, cell types, and culture characteristics of chicken small intestinal organoids were studied by HE staining, transmission electron microscopy, reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), indirect immunofluorescence, and immunohistochemistry. The results showed that the 3D organoids of the chicken small intestine cultured in vitro were morphologically consistent with the chicken intestinal tissue and contained differentiated epithelial cells. In summary, we successfully established an in vitro culture system for chicken small intestinal organoids, providing a new method for the subsequent research on chicken intestinal physiology, pathology, and host-pathogen interaction mechanism and the development of relevant drugs.
Animals ; Organoids/metabolism* ; Intestine, Small/drug effects* ; Chickens ; Cell Culture Techniques/methods* ; Culture Media ; Chick Embryo ; Tissue Culture Techniques/methods*

Cognitive impairment is one of the core symptoms of schizophrenia, and there is no recognized effective treatment for it. Current studies have found that treatment targeting the N-methyl-D-aspartate receptor (NMDAR) can improve cognitive function in patients with schizophrenia. This article reviewed the research progress of NMDAR-related modulators in improving cognitive impairment of schizophrenia, summarized the application of D-serine, glycine, D-cycloserine, memantine, etc., and discussed the current research deficiencies and future research directions .

Cognitive impairment is one of the core symptoms of schizophrenia, and there is no recognized effective treatment for it. Current studies have found that treatment targeting the N-methyl-D-aspartate receptor (NMDAR) can improve cognitive function in patients with schizophrenia. This article reviewed the research progress of NMDAR-related modulators in improving cognitive impairment of schizophrenia, summarized the application of D-serine, glycine, D-cycloserine, memantine, etc., and discussed the current research deficiencies and future research directions .

Objective To investigate the value of Charlson comorbidity index (CCI) in predicting the short- and long-term risks of death in patients with acute-on-chronic liver failure (ACLF). Methods A total of 317 patients with ACLF who attended The First Hospital of Lanzhou University from December 1, 2016 to December 1, 2021 were enrolled, and according to their prognosis, they were divided into death group with 169 patients and survival group with 148 patients. The two groups were analyzed in terms of clinical data and follow-up data. The group t -test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. The univariate and multivariate Cox proportional-hazards regression model analyses were used to investigate the influencing factors for the prognosis of ACLF patients. The Kaplan-Meier method was used to plot survival curves, and the Log-rank test was used for comparison of survival time between patients with different CCI scores. The receiver operating characteristic (ROC) curve was used to evaluate the performance of CCI and other indices in assessing the prognosis of ACLF patients. Results Among the 317 patients, there were 225 (71.0%) male patients. There were significant differences between the death group and the survival group in age, hemoglobin, white blood cell count, total bilirubin, albumin, Model for End-Stage Liver Disease (MELD) score, prothrombin time activity, CCI, age-adjusted Charlson co-morbidity index (ACCI), and follow-up time (all P < 0.05). The multivariate Cox regression analysis showed that the CCI (hazard ratio [ HR ]=1.351, 95% confidence interval [ CI ]: 1.112-1.641, P =0.002), ACCI ( HR =1.200, 95% CI : 1.011-1.423, P =0.037), and MELD score ( HR =1.076, 95% CI : 1.054-1.099, P < 0.001) were independent risk factors for the prognosis of ACLF patients. Based on CCI score, the patients were divided into CCI ≤4 group with 167 patients, CCI=5 group with 64 patients, and CCI ≥6 group with 86 patients, with a 3-year mortality rate of 26.5%, 83.2%, and 96.9%, respectively, and there was a significant difference in survival time between any two groups after 3 years of follow-up and at the time of follow-up till September 2022 (all P < 0.001). CCI, ACCI, and MELD scores had an area under the ROC curve of 0.845, 0.811, and 0.790, respectively, in predicting the prognosis of ACLF patients. Conclusion As commonly used comorbidity assessment indices, CCI and ACCI scores have certain value in evaluating the short- and long-term prognosis of ACLF patients.

Objective:To analyze the application and regularity of acupoint selection of Sanyinjiao (SP 6) based on data mining.Methods:Search for literatures in CNKI, Wanfang, VIP, and Pubmed, the clinical researches of acupuncture on Sanyinjiao (SP 6) point were selected, according to the inclusion and exclusion criteria, and the retrieval period was from database construction to September 30th, 2021. Excel 2016, SPSS Statistics 25.0, SPSS Modeler 18.0 were used to perform descriptive analysis, association analysis and cluster analysis.Results:After literature screening, a total of 261 literatures were included, involving 73 kinds of diseases, mainly including mental and behavioral disorders, genitourinary diseases, endocrine and nutritional metabolism diseases and nervous system diseases. The most frequently used acupoints in Sanyinjiao (SP 6) compatibility are Zusanli (ST 36), Baihui (GV 20), Guanyuan (CV 4) and Taichong (LR 3), most of which focus on stomach meridian, conception channel, governor channel and bladder meridian. Seven categories were extracted among high-frequency acupoints by cluster analysis. The association rule analysis showed that the commonly used combination of Sanyinjiao (SP 6) were Zusanli (ST 36)-Sanyinjiao (SP 6), Baihui (GV 20)-Sanyinjiao (SP 6), and Guanyuan (CV 4)-Sanyinjiao (SP 6).Conclusions:Sanyinjiao (SP 6) is widely used in clinical application, and it is always compatible with stomach meridian, conception vessel, governor channel acupoints, especially those acupoints on the outer and inner meridians and the upper and lower parts. Sanyinjiao (SP 6) combined with other acupoints can treat diseases of multiple systems, such as insomnia, stroke, anxiety and depression, dysmenorrhea, infertility, etc. Clustering and association analysis found the core compatibility law of Sanyinjiao (SP 6), which can be used as a reference for clinical acupoint selection.

Primary sclerosing cholangitis （PSC） is an immune-mediated chronic cholestatic liver disease and can progress to end-stage liver diseases such as liver cirrhosis and liver failure， and there are still no effective treatment methods at present. Studies have found that T lymphocytes are closely associated with the development and progression of PSC. This article reviews the role of T lymphocytes in PSC， so as to provide new ideas for research on the pathogenesis of PSC and the clinical diagnosis and treatment of PSC.