Objective: To investigate the factors affecting drug-induced liver injury among patients with pulmonary tuberculosis in Ningbo City from 2015 to 2019, so as to provide insights into the prevention of drug-induced liver injury. Methods: Demographic features, presence of drug-induced liver injury, and disease history prior to anti-tuberculosis therapy were captured from patients with pulmonary tuberculosis in Ningbo City from 2015 to 2019 through the Tuberculosis Management Information System of the Chinese Disease Control and Prevention Information System and Ningbo Regional Diagnosis and Treatment Information Platform. Factors affecting drug-induced liver injury was identified using the multivariable logistic regression analysis. Results: A total of 9 397 patients with pulmonary tuberculosis were enrolled, among whom 66.43% ( 6 242 case ) were male, 65.89% ( 6 192 cases ) were at ages of <60 years, and 92.35% ( 8 678 cases ) were treatment-naïve. There were 1 425 patients with drug-induced liver injury (15.16% incidence), including 729 cases with grade 1 (51.16%), 24 cases with grade 2 (1.68%), 7 cases with grade 3 (0.49%), 7 cases with grade 4 ( 0.49% ), and 658 cases with ungraded drug-induced liver injury ( 46.18% ). The median duration between drug administration and development of drug-induced liver injury was 24 ( interquartile range, 44 ) days. Multivariable logistic regression analysis identified treatment-naïve ( OR=1.464, 95%CI: 1.153-1.859 ) and history of liver disease ( OR=2.001, 95%CI: 1.709-2.342) as risk factors for drug-induced liver injury in patients with pulmonary tuberculosis. Conclusion The incidence of drug-induced liver injury was 15.16% among pulmonary tuberculosis patients in Ningbo City from 2015 to 2019. Treatment-naïve and a history of liver disease are associated with drug-induced liver injury among patients with pulmonary tuberculosis.

The conventional analytical methods cannot effectively adjust for time-varying confounding that occur in a longitudinal study and thus cannot correctly estimate the causal effects. This study explains the necessity of precisely controlling time-varying confounding and outlines G methods, including parametric g-formula, inverse probability of weighting, and G-estimation. We also compare the methods above to provide a reference for correctly estimating causal effects in the longitudinal study.

[摘 要] CD8+ T细胞是抗肿瘤免疫应答的主要执行者。通过重塑CD8+ T细胞杀伤肿瘤细胞的能力，免疫疗法已在抗肿瘤领域取得重大突破，但临床获益仅局限于部分患者和癌症类型。如何克服CD8+ T细胞功能障碍是肿瘤免疫疗法亟待解决的关键问题。近年来，多项研究揭示了CD8+ T细胞的干性调控机制，发现了干细胞样CD8+ T细胞具有自我更新和增殖能力，阐明了该细胞亚群在维持持续性肿瘤免疫治疗应答中的重要性。本文论述了干细胞样CD8+ T细胞的分子与功能特征、CD8+ T细胞干性的细胞内外影响因素，归纳总结了目前靶向CD8+ T细胞的干性重编程策略，进一步展望了靶向CD8+ T细胞干性程序来提高肿瘤免疫疗法疗效的思路和方法。

Objective To develop a remote diabetic retinopathy (DR) screening system and to evaluate the effectiveness of the screening system in community.Methods A cross-sectional study was carried out under the informed consent of subjects in Peking University People's Hospital and Beijing Xicheng District Desheng Community Health Service Center from June 2015 to December 2016.A remote DR screening system was established in Peking University People's Hospital and Beijing Xicheng District Desheng Community Health Service Center during June 2015 to December 2016.Based on non-mydriatic digital eye fundus camera photography and the internet transmission technology,anterior ocular segment and fundus images of 2 473 eyes from 1 355 community subjects with type 2 diabetes mellitus were transmitted from Beijing Xicheng District Desheng Community Health Service Center to the reading center of Peking University People's Hospital,and the results were provided to the subjects after analysis,including visual examination,diagnosis and follow-up rate of the subjects,the agreement between remote screening system and conventional screening method was analyzed and compared.Results The visual acuities of the 2 473 eyes of 1 355 subjects were obtained by trained community physician,and the visual acuity was ≤0.05 in 103 eyes (4.2％),＞0.05-0.3 in 780 eyes (31.5％),＞0.3 in 1 590 eyes (64.3％).A good consistency was found in the diagnosis and grading of DR (Kappa value =0.895) and in diagnosis of macular disorder (Kappa value =0.763)between the remote screening system and conventional screening method.In addition,the diagnosis results of retinal photocoagulation were consistent between the two methods (Kappa value =1.000).The mean duration of the remote screening system for one subject was 10 minutes,which was shorter than 23 minutes of conventional screening method.The follow-up rate of remote screening system was 75.2％.Conclusions There is a high consistency in the DR diagnosis and evaluation between the remote non-mydriatic screening system and conventional screening method.The screening program with follow-up requests has a satisfying follow-up rate,which could meet the demand of DR screening.

Objective To investigate the role of glutamatergic neurons in the dorsomedial periaqueductal grey(dmPAG)in regulating excessive defensive behaviors in mice with post-traumatic stress disorder(PTSD).Methods Eight-week-old male C57BL/6 mice were subjected to stereotactic injections of different recombinant adeno-associated viral vectors(rAAV2/9-CaMKⅡ-mCherry,rAAV2/9-CaMKⅡ-hM3Dq-mCherry and rAAV2/9-CaMKⅡ-hM4Di-mCherry)into the bilateral dmPAG for chemogenetic activation or inhibition of the glutamatergic neurons,followed 2 weeks later by PTSD modeling by single prolonged stress.The looming test,response to whisker stimulation test and contextual fear conditioning(CFC)test were used to observe changes in defensive behaviors of the PTSD mice.The activity of glutamatergic neurons in the dmPAG were observed using immunofluorescence staining.Results Compared with the control mice,the mouse models of PTSD showed a shortened latency of flights with increased time spent in the nest,response scores of defensive behaviors and freezing time(all P<0.01).Immunofluorescence staining revealed significantly increased c-fos-positive glutamatergic neurons in the dmPAG of PTSD mice with defensive behaviors.Activation of the glutamatergic neurons in the dmPAG(in PTSD hM3Dq group)did not cause significant changes in the latency of flights or time in nest but obviously increased response scores of defensive behaviors and freezing time of the mice,whereas inhibiting the glutamatergic neurons in the dmPAG(in PTSD hM4Di group)caused the reverse changes and obviously alleviated defensive behaviors in the PTSD mice(P<0.05 or 0.01).Conclusion Inhibiting the activity of glutamatergic neurons in the dmPAG can alleviate defensive behaviors in mice with PTSD.

Objective To investigate the role of glutamatergic neurons in the dorsomedial periaqueductal grey(dmPAG)in regulating excessive defensive behaviors in mice with post-traumatic stress disorder(PTSD).Methods Eight-week-old male C57BL/6 mice were subjected to stereotactic injections of different recombinant adeno-associated viral vectors(rAAV2/9-CaMKⅡ-mCherry,rAAV2/9-CaMKⅡ-hM3Dq-mCherry and rAAV2/9-CaMKⅡ-hM4Di-mCherry)into the bilateral dmPAG for chemogenetic activation or inhibition of the glutamatergic neurons,followed 2 weeks later by PTSD modeling by single prolonged stress.The looming test,response to whisker stimulation test and contextual fear conditioning(CFC)test were used to observe changes in defensive behaviors of the PTSD mice.The activity of glutamatergic neurons in the dmPAG were observed using immunofluorescence staining.Results Compared with the control mice,the mouse models of PTSD showed a shortened latency of flights with increased time spent in the nest,response scores of defensive behaviors and freezing time(all P<0.01).Immunofluorescence staining revealed significantly increased c-fos-positive glutamatergic neurons in the dmPAG of PTSD mice with defensive behaviors.Activation of the glutamatergic neurons in the dmPAG(in PTSD hM3Dq group)did not cause significant changes in the latency of flights or time in nest but obviously increased response scores of defensive behaviors and freezing time of the mice,whereas inhibiting the glutamatergic neurons in the dmPAG(in PTSD hM4Di group)caused the reverse changes and obviously alleviated defensive behaviors in the PTSD mice(P<0.05 or 0.01).Conclusion Inhibiting the activity of glutamatergic neurons in the dmPAG can alleviate defensive behaviors in mice with PTSD.

Objective:To investigate the effect of tofacitinib,a pan-Janus kinase(JAK)inhibitor,on transforming growth factor-beta 1(TGF-β1)-induced fibroblast to myofibroblast transition(FMT)and to explore its mechanism.To provide a theoretical basis for the clinical treatment of connective tissue disease-related interstitial lung disease(CTD-ILD).Methods:(1)Human fetal lung fibroblast 1(HFL-1)were cultured in vitro,and 6 groups were established:DMSO blank control group,TGF-β1 in-duction group,and TGF-β1 with different concentrations of tofacitinib(0.5,1.0,2.0,5.0 μmol/L)drug intervention experimental groups.CCK-8 was used to measure the cell viability,and wound-healing assay was performed to measure cell migration ability.After 48 h of combined treatment,quantitative real-time PCR(RT-PCR)and Western blotting were used to detect the gene and protein expression levels of α-smooth muscle actin(α-SMA),fibronectin(FN),and collagen type Ⅰ(COL1).(2)RT-PCR and enzyme-linked immunosorbnent assay(ELISA)were used to detect the interleukin-6(IL-6)gene and protein expression changes,respectively.(3)DMSO carrier controls,1.0 μmol/L and 5.0 μmol/L tofacitinib were added to the cell culture media of different groups for pre-incubation for 30 min,and then TGF-β1 was added to treat for 1 h,6 h and 24 h.The phosphorylation levels of Smad2/3 and signal transducer and activator of transcription 3(STAT3)protein were detected by Western blotting.Results:(1)Tofacitinib inhibited the viability and migration ability of HFL-1 cells after TGF-β1 induction.(2)The expression of α-SMA,COL1A1 and FN1 genes of HFL-1 in the TGF-β1-induced groups was signifi-cantly up-regulated compared with the blank control group(P＜0.05).Compared with the TGF-β1 in-duction group,α-SMA expression in the 5.0 μmol/L tofacitinib intervention group was significantly inhi-bited(P＜0.05).Compared with the TGF-β1-induced group,FN1 gene was significantly inhibited in each intervention group at a concentration of 0.5-5.0 μmol/L(P＜0.05).Compared with the TGF-β1-induced group,the COL1A1 gene expression in each intervention group did not change significantly.(3)Western blotting results showed that the protein levels of α-SMA and FN1 in the TGF-β1-induced group were significantly higher than those in the control group(P＜0.05),and there was no significant difference in the expression of COL1A1.Compared with the TGF-β1-induced group,the α-SMA protein level in the intervention groups with different concentrations decreased.And the differences between the TGF-β1-induced group and 2.0 μmol/L or 5.0 μmol/L intervention groups were statistically significant(P＜0.05).Compared with the TGF-β1-induced group,the FN1 protein levels in the intervention groups with different concentrations showed a downward trend,but the difference was not statistically sig-nificant.There was no difference in COL1A1 protein expression between the intervention groups com-pared with the TGF-β1-induced group.(4)After TGF-β1 acted on HFL-1 cells for 48 h,the gene ex-pression of the IL-6 was up-regulated and IL-6 in culture supernatant was increased,the intervention with tofacitinib partly inhibited the TGF-β1-induced IL-6 gene expression and IL-6 in culture supernatant.TGF-β1 induced the increase of Smad2/3 protein phosphorylation in HFL-1 cells for 1 h and 6 h,STAT3 protein phosphorylation increased at 1 h,6 h and 24 h,the pre-intervention with tofacitinib inhibited the TGF-β1-induced Smad2/3 phosphorylation at 6 h and inhibited TGF-β1-induced STAT3 phosphorylation at 1 h,6 h and 24 h.Conclusion:Tofacitinib can inhibit the transformation of HFL-1 cells into myofi-broblasts induced by TGF-β1,and the mechanism may be through inhibiting the classic Smad2/3 path-way as well as the phosphorylation of STAT3 induced by TGF-β1,thereby protecting the disease progres-sion of pulmonary fibrosis.

Objective:To study the epidemiological and clinical characteristics of brucellosis in Xinjiang Uygur Autonomous Region.Methods:A retrospective analysis method was used to collect medical records of 581 patients with brucellosis who visited the Xinjiang Uygur Autonomous Regional People's Hospital from January 2009 to December 2019. Demographic and epidemiological characteristics, clinical symptoms and signs, and laboratory test results of the patients were analyzed.Results:Among 581 patients with brucellosis, the male to female ratio was 2.8 ∶ 1.0 (428 ∶ 153). The age was (44.41 ± 16.25) years old, ranging from 1 - 83 years old, and mainly concentrated in 35 - 60 years old, accounting for 70.91% (412/581). The ethnic distribution was dominated by Uyghur, accounting for 50.60% (294/581). The occupational distribution was mainly farmers, accounting for 43.20% (251/581). A total of 186 patients had a clear history of contact with cattle and sheep, accounting for 32.01% (186/581). The clinical stage was dominated by patients in the acute stage, accounting for 55.25% (321/581). There were 48 cases of complications, accounting for 8.26%(48/581). The main clinical symptom of brucellosis patients was pain and fever, accounting for 73.67% (428/581) and 66.61% (387/581), respectively. Laboratory tests were dominated by increased blood sedimentation and C-reactive protein, accounting for 29.09% (169/581) and 23.06% (134/581), respectively. The positive rate of Brucella culture was low, accounting for 4.48% (26/581). Conclusions:The majority of brucellosis patients in Xinjiang Uygur Autonomous Region are young and middle-aged males, with the main occupation being farmers. The clinical symptoms are mostly pain and fever. The positive rate of Brucella culture in patients is relatively low. It is recommended to combine epidemiological and clinical features for diagnosis to reduce missed diagnosis and misdiagnosis, and detect and treat it early.

OBJECTIVE: To explore the regulatory effects of GABAergic neurons in the zona incerta (ZI) on sevoflurane and propofol anesthesia. METHODS: Forty-eight male C57BL/6J mice divided into 8 groups (n=6) were used in this study. In the study of sevoflurane anesthesia, chemogenetic experiment was performed in 2 groups of mice with injection of either adeno-associated virus carrying hM3Dq (hM3Dq group) or a virus carrying only mCherry (mCherry group). The optogenetic experiment was performed in another two groups of mice injected with an adeno-associated virus carrying ChR2 (ChR2 group) or GFP only (GFP group). The same experiments were also performed in mice for studying propofol anesthesia. Chemogenetics or optogenetics were used to induce the activation of GABAergic neurons in the ZI, and their regulatory effects on anesthesia induction and arousal with sevoflurane and propofol were observed; EEG monitoring was used to observe the changes in sevoflurane anesthesia maintenance after activation of the GABAergic neurons. RESULTS: In sevoflurane anesthesia, the induction time of anesthesia was significantly shorter in hM3Dq group than in mCherry group (P < 0.05), and also shorter in ChR2 group than in GFP group (P < 0.01), but no significant difference was found in the awakening time between the two groups in either chemogenetic or optogenetic tests. Similar results were observed in chemogenetic and optogenetic experiments with propofol (P < 0.05 or 0.01). Photogenetic activation of the GABAergic neurons in the ZI did not cause significant changes in EEG spectrum during sevoflurane anesthesia maintenance. CONCLUSION Activation of the GABAergic neurons in the ZI promotes anesthesia induction of sevoflurane and propofol but does not affect anesthesia maintenance or awakening.
Male ; Animals ; Mice ; Mice, Inbred C57BL ; Propofol/pharmacology* ; Sevoflurane/pharmacology* ; Zona Incerta ; Anesthesia, General ; GABAergic Neurons

The cochlear auditory epithelium contains two types of sound receptors, inner hair cells (IHCs) and outer hair cells (OHCs). Mouse models for labelling juvenile and adult IHCs or OHCs exist; however, labelling for embryonic and perinatal IHCs or OHCs are lacking. Here, we generated a new knock-in Fgf8^P2A-3×GFP/+ (Fgf8^GFP/+) strain, in which the expression of a series of three GFP fragments is controlled by endogenous Fgf8 cis-regulatory elements. After confirming that GFP expression accurately reflects the expression of Fgf8, we successfully obtained both embryonic and neonatal IHCs with high purity, highlighting the power of Fgf8^GFP/+. Furthermore, our fate-mapping analysis revealed, unexpectedly, that IHCs are also derived from inner ear progenitors expressing Insm1, which is currently regarded as an OHC marker. Thus, besides serving as a highly favorable tool for sorting early IHCs, Fgf8^GFP/+ will facilitate the isolation of pure early OHCs by excluding IHCs from the entire hair cell pool.
Animals ; Mice ; Hair Cells, Auditory, Inner ; Cochlea/metabolism* ; Hair Cells, Auditory, Outer/metabolism* ; Disease Models, Animal ; Fibroblast Growth Factor 8/metabolism*