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.

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*

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.

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.

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 efficacy and safety of the 12-week regimen with sofosbuvir and coblopasvir hydrochloride in the treatment of chronic hepatitis C (CHC) in northwest China. Methods This study enrolled 101 patients with CHC of any genotype who received sofosbuvir (400 mg) combined with coblopasvir hydrochloride (60 mg) for 12 weeks in The First Affiliated Hospital of Air Force Medical University, The Second Affiliated Hospital of Air Force Medical University, The Second Affiliated Hospital of Xi'an Jiaotong University, and Baoji Central Hospital from July 1 to December 31, 2021, among whom 13 had liver cirrhosis and 88 did not have live cirrhosis. Other antiviral drugs such as ribavirin were not added regardless of the presence or absence of liver cirrhosis or the genotype of CHC. Related clinical data ere extracted, including HCV RNA quantification and liver biochemical parameters at baseline, at week 12 of treatment, and at 12 weeks after drug withdrawal. The primary endpoints were sustained virologic response at 12 weeks after the end of treatment (SVR12) and safety at week 12 of treatment, and the secondary endpoint was the effect of the 12-week treatment on liver biochemical parameters. The non-normally distributed continuous data were expressed as M ( P 25 - P 75 ), and the Mann-Whitney U test was used for comparison between groups. Results A total of 101 patients were included in the analysis, among whom there were 55 male patients (54.5%) and 46 female patients, and the median age was 53 years. Among these patients, 12.8% had liver cirrhosis, 1.0% had liver cancer, 3.0% were treatment-experienced patients, and 3.0% had type 2 diabetes. As for genotype distribution, 8% had CHC genotype 1, 60% had CHC genotype 2, 19% had CHC genotype 3, and 6% had CHC genotype 6, and genotype was not tested for 7% of the patients. After 12 weeks of treatment, all 101 patients had a HCV RNA level of below the lower limit of detection and an SVR12 rate of 100%, with a significant reduction in the serum level of alanine aminotransferase (ALT) from baseline to week 12 of treatment ( P < 0.05). Among these patients, 22.7% had concomitant medications such as atorvastatin calcium, aspirin, metformin, nifedipine, bicyclol, and compound glycyrrhizin. The incidence rate of adverse events was 16.8%, and fatigue (12.9%) was the most common adverse event. Conclusion The 12-week treatment with sofosbuvir and coblopasvir hydrochloride can obtain high SVR12 in CHC patients in northwest China and has good antiviral safety, with a significant improvement in abnormal serum ALT at week 12 of treatment.

Objective:To analyze the clinical and laboratory characteristics of mpox patients in Guangzhou City.Methods:The general conditions, symptoms and signs, and laboratory test results of mpox patients admitted to Guangzhou Eighth People′s Hospital, Guangzhou Medical University from June 8 to June 21, 2023 were collected. The clinical characteristics of human immunodeficiency virus (HIV)-infected patients and HIV-negative patients were compared.Independent sample t test, Mann-Whitney U test and Fisher exact probability method were used for statistical analysis. Results:Nineteen mpox patients were included in this study, none of them had been vaccinated with smallpox vaccine. All of them were identifed as gay men with an age of (33.2±6.4) years. And all of them had sex with men within 21 days of onset. There were eight cases with HIV infection and syphilis, respectively. All of these 19 patients had skin lesions which were the first symptom of 15 patients, and appeared during the course of the disease of four patients. Other common symptoms and signs were lymph node enlargement (17 cases), lymph node tenderness (15 cases), pharyngeal congestion (15 cases), tonsil enlargement (13 cases), fever (11 cases) and pruritus (10 cases). There were 13 cases with elevated CD8 + T lymphocytes, interleukin-10 level and procalcitonin level, respectively, 12 cases with elevated serum amyloid A level, and seven cases with elevated blood lymphocyte count. Eight patients were infected with HIV, including one acute infection and seven chronic infections (all of them had received regular antiviral therapy and had a CD4 + T lymphocyte count of 657(400, 757)/μL before onset). There were no significant differences in age ( t=1.55), incidence of complications (Fisher exact probability method), number of skin lesions ( Z=-0.21), incidence of lymph node enlargement (Fisher exact probability method), incidence of fever (Fisher exact probability method), duration of fever ( Z=-0.48), lymphocyte count ( t=-1.55), CD4 + T lymphocyte count ( Z=-0.17) and CD8 + T lymphocyte count ( Z=-1.49) between the HIV-infected patients and HIV-negative patients (all P>0.05). The number of skin lesions was 19(6, 26), and the locations of skin lesions were mainly in perineum and nearby areas (17 cases). The most frequent morphology of skin lesions at admission were papules (15 cases), eschar (15 cases) and pustules (12 cases). All patients recovered after topical medication and symptomatic treatment. Conclusions:Mpox mainly occurs in young and middle-aged men who have sex with men in Guangzhou City. Skin lesions, lymph node enlargement accompanied by tenderness, pharyngeal congestion, tonsil enlargement and fever are the most common features. Common laboratory abnormities are elevated inflammatory markers. The clinical characteristics of HIV-infected patients with normal immune function are similar to those of HIV-negative patients.

Programmed cell death (PCD) is a genetically determined, active and orderly cell death in the organism, and it affects the evolution of the organism, maintenance of its homeostasis, and development of several tissues and organs. The abnormal regulation of this process is closely related to various human diseases, including cancer. The identified pathways of PCD include apoptosis, autophagy, necroptosis, pyroptosis, and ferroptosis, which can be activated when cells are stimulated by various internal and external environmental factors. These pathways can induce cell death or maintain cell survival in kidney cancer cells under the regulation of various signaling molecules, thus affecting tumor progression or therapeutic efficacy. In this paper, the role of these PCD pathways in the development of kidney cancer was reviewed in light of recent research advances to provide new directions for the in-depth study of the pathogenesis of kidney cancer and the development of targeted antitumor drugs.