A spatiotemporal atlas of cholestatic injury and repair in mice.

Cholestatic liver injuries, characterized by regional damage around the bile ductular region, lack curative therapies and cause considerable mortality. Here we generated a high-definition spatiotemporal atlas of gene expression during cholestatic injury and repair in mice by integrating spatial enhanced resolution omics sequencing and single-cell transcriptomics. Spatiotemporal analyses revealed a key role of cholangiocyte-driven signaling correlating with the periportal damage-repair response. Cholangiocytes express genes related to recruitment and differentiation of lipid-associated macrophages, which generate feedback signals enhancing ductular reaction. Moreover, cholangiocytes express high TGFß in association with the conversion of liver progenitor-like cells into cholangiocytes during injury and the dampened proliferation of periportal hepatocytes during recovery. Notably, Atoh8 restricts hepatocyte proliferation during 3,5-diethoxycarbonyl-1,4-dihydro-collidin damage and is quickly downregulated after injury withdrawal, allowing hepatocytes to respond to growth signals. Our findings lay a keystone for in-depth studies of cellular dynamics and molecular mechanisms of cholestatic injuries, which may further develop into therapies for cholangiopathies.

A spatiotemporal atlas of mouse liver homeostasis and regeneration.

The mechanism by which mammalian liver cell responses are coordinated during tissue homeostasis and perturbation is poorly understood, representing a major obstacle in our understanding of many diseases. This knowledge gap is caused by the difficulty involved with studying multiple cell types in different states and locations, particularly when these are transient. We have combined Stereo-seq (spatiotemporal enhanced resolution omics-sequencing) with single-cell transcriptomic profiling of 473,290 cells to generate a high-definition spatiotemporal atlas of mouse liver homeostasis and regeneration at the whole-lobe scale. Our integrative study dissects in detail the molecular gradients controlling liver cell function, systematically defining how gene networks are dynamically modulated through intercellular communication to promote regeneration. Among other important regulators, we identified the transcriptional cofactor TBL1XR1 as a rheostat linking inflammation to Wnt/ß-catenin signaling for facilitating hepatocyte proliferation. Our data and analytical pipelines lay the foundation for future high-definition tissue-scale atlases of organ physiology and malfunction.

Cell atlas of CCl 4-induced progressive liver fibrosis reveals stage-specific responses.

Chronic liver injury leads to progressive liver fibrosis and ultimately cirrhosis, a major cause of morbidity and mortality worldwide. However, there are currently no effective anti-fibrotic therapies available, especially for late-stage patients, which is partly attributed to the major knowledge gap regarding liver cell heterogeneity and cell-specific responses in different fibrosis stages. To reveal the multicellular networks regulating mammalian liver fibrosis from mild to severe phenotypes, we generated a single-nucleus transcriptomic atlas encompassing 49 919 nuclei corresponding to all main liver cell types at different stages of murine carbon tetrachloride (CCl 4)-induced progressive liver fibrosis. Integrative analysis distinguished the sequential responses to injury of hepatocytes, hepatic stellate cells and endothelial cells. Moreover, we reconstructed cell-cell interactions and gene regulatory networks implicated in these processes. These integrative analyses uncovered previously overlooked aspects of hepatocyte proliferation exhaustion and disrupted pericentral metabolic functions, dysfunction for clearance by apoptosis of activated hepatic stellate cells, accumulation of pro-fibrotic signals, and the switch from an anti-angiogenic to a pro-angiogenic program during CCl 4-induced progressive liver fibrosis. Our dataset thus constitutes a useful resource for understanding the molecular basis of progressive liver fibrosis using a relevant animal model.

Efficacy of Modified Huangqi Chifeng decoction in alleviating renal fibrosis in rats with IgA nephropathy by inhibiting the TGF-ß1/Smad3 signaling pathway through exosome regulation.

ETHNOPHARMACOLOGICAL RELEVANCE: IgA nephropathy is the most common form of primary glomerulonephritis and is a major cause of renal failure worldwide. Modified Huangqi Chifeng decoction (MHCD), a traditional Chinese herbal preparation, has clinical efficacy in reducing the 24-h urine protein levels in patients with IgA nephropathy. However, the molecular mechanism of MHCD needs further study. AIM OF THE STUDY: This study aimed to investigate the mechanisms by which MHCD treatment alleviates renal fibrosis. MATERIALS AND METHODS: An IgA nephropathy rat model was established using bovine serum albumin, carbon tetrachloride, and lipopolysaccharide. The rats were divided into control, model, telmisartan, low-dose MHCD, medium-dose MHCD, and high-dose MHCD groups. Treatments were administered to these groups for 8 weeks. Subsequently, the 24-h urine protein, serum creatinine, blood urea nitrogen, and blood albumin levels were measured. Pathological changes and degree of fibrosis in renal tissues were observed, and levels of the transforming growth factor-ß1 (TGF-ß1)/Smad3 signaling pathway components in renal tissues and TGF-ß1 in urinary exosomes were measured. RESULTS: Telmisartan and MHCD reduced 24-h urine protein levels, alleviated renal pathological injury, and decreased the renal expression of fibronectin, laminin, and collagen IV in rats with IgA nephropathy. Urinary exosomes were extracted and identified for further investigation of their role in renal fibrosis. MHCD reduced TGF-ß1 expression in urinary exosomes and reduced TGF-ß1 and p-Smad3 levels in renal tissues. CONCLUSION: MHCD alleviated renal fibrosis in rats with IgA nephropathy by inhibiting the TGF-ß1/Smad3 signaling pathway through the downregulation of TGF-ß1 expression in exosomes.

A localized small-scale external quality assessment (EQA) for PCR testing of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the molecular laboratories.

The aim of this study was to estimate the PCR results for SARS-CoV-2 testing in 32 participating laboratories in a localized small-scale external quality assessment (EQA) scheme. EQA samples were distributed to the participants and detected immediately on the day of delivery. Qualitative results were submitted to the EQA provider, including negative or positive results along with cycle threshold (Ct) values for different target genes. Although the variability of Ct values differed among the laboratories in the EQA, a total of 32 (100 %) participants reported correct qualitative results. The study showed that the mean loads of N or E gene were higher than those of ORF1ab in SARS-CoV-2 RNA samples. Regardless of the analyzed gene target, the mean Ct values for weak positive and positive samples varied by fewer than 1.74 and 1.91 cycles, respectively. Less than 12 % of reported Ct values for ORF1ab and N genes deviated by more than ±4 cycles (maximum: -9.92 cycles), while none deviated by more than ±4 cycles for the E gene. The current EQA program can provide a robust practical basis for follow-up planning to conduct evaluations for SARS-CoV-2 PCR testing and other novel emerging pathogens in the future.

Serum carbohydrate antigen 72-4 concentrations decrease with age in females but not in males in Beijing, China.

OBJECTIVE: Carbohydrate antigen 72-4 (CA72-4) is widely used in the diagnosis and monitoring of many cancers. However, there are few studies on the differences of CA72-4 concentrations in terms of age and gender. METHODS: A total of 10,957 healthy subjects were divided into two groups according to gender and three age groups. The serum CA72-4 were detected. Statistical analysis was performed by SPSS. RESULTS: The CA72-4 concentration in female group was significantly higher than that in male group. The concentration of CA72-4 gradually decreased with age. Compared with the age >60 group, the CA72-4 concentrations were increased in the age 46-60 group and 16-45 group (P >0.05, respectively). To better observe the age difference, the age 16-45 and 46-60 groups were combined into the age 16-60 group. In comparison to the age >60 group, the CA72-4 concentration of age 16-60 group was significantly increased (P = 0.000). In the age >60 group, there was no difference between genders. Nevertheless, the difference between the sexes in the age 16-60 group was significant (P = 0.023). CONCLUSIONS: The reference interval of CA72-4 for local healthy population was established. CA72-4 concentrations gradually decreased with the increase of age, and CA72-4 concentration in females aged 16-60 years (0-18.0 U/mL) was higher than in males (0-14.5 U/mL); however, there was no gender difference in the age group above 60 years old (0-14.5 U/mL). Moreover, in male CA72-4 there was no significant difference among all age groups, while the potential mechanism of female changes with age needs further study.

Clinician-collected urethra swab: A good alternative sample type using cobas 4800 system for Chlamydia trachomatis and Neisseria gonorrhoeae in men.

BACKGROUND: Nucleic acid amplification tests (NAATs) are being used increasing to detection of CT (Chlamydia trachomatis) and NG (Neisseria gonorrhoeae) infections for superior sensitivity and specificity than other tests. Male first-void urine (FVU) sample is the optimal sample type for detection of CT and NG by NAATs. Although not being the recommended by NAATs, clinician-collected urethra swab (CCUS) is perhaps a good alternative sample type compared with the FVU sample in men. METHODS: Paired samples (FVU and CCUS) from one hundred male outpatients were simultaneously detected by urine pattern and swab pattern using cobas 4800 CT/NG assay on cobas 4800 system for the detection of CT and NG, respectively. And twenty-one positive controls were also detected on cobas 4800 system. RESULTS: The CT/NG cycle thresholds (Ct) value of urine pattern is lower than that of swab pattern for the same positive samples (clinical samples and positive controls) on the cobas 4800 CT/NG assay. The final CT/NG results of two sample patterns from patients were highly consistent except for four discordant results. CONCLUSION: CCUS is validated for a good alternative sample type for the CT/NG detection on the cobas 4800 system in this study.

The Effects of Matrine in Combination with Docetaxel on Castration-Resistant (Androgen-Independent) Prostate Cancer.

BACKGROUND: Matrine (MAT) exhibits higher efficacy of chemotherapy when it is combined with other chemotherapeutic drugs; however, the therapeutic efficacy of matrine in combination with docetaxel (DOC) for prostate cancer, or even androgen-independent prostate cancer, remains poorly understood and the underlying molecular mechanisms have not yet been clearly defined. In the present study, we investigated whether matrine combined with docetaxel can strengthen anti-cancer effect. METHODS: In this study, 7 groups were established, including (1) blank control group (cells). (2) 0.1 g/L MAT group, (3) 0.5 g/L MAT group, (4) 0.1 g/L MAT+ 50 µg/L DOC group, (5) 0.5 g/L MAT+ 50 µg/L DOC group, (6) 0.1 g/L MAT+ 100 µg/L DOC group, and (7) 0.5 g/L MAT+ 100 µg/L DOC group. MTS assay was performed to detect the anti-proliferative effects of each group on DU145 and PC-3 cells. At the same time, Transwell assay was performed to detect anti-migrative and anti-invasive effects of each group on DU145 and PC-3 cells. Biochemical colorimetric method and enzyme-linked immunosorbent assay were performed to detect the levels of LDH, IL-1ß and IL-18 of each group on DU145 and PC-3 cells. Flow cytometry (FCM) assay was used to do the apoptosis analysis on DU145 and PC-3 cells of each group. At last, Western blot analysis was performed to investigate the expression levels of caspase1 in cells of each group. Statistical analyses were performed with SPSS 17.0 (SPSS Inc, USA) software, and one-way ANOVA and Fisher's exact test was taken. RESULTS: MTS assay showed that matrine combined with docetaxel could inhibit both DU145 and PC-3 cells' proliferation in a dose- and time-dependent manner. Transwell assay showed that matrine combined with docetaxel could inhibit both DU145 and PC-3 cells' migration and invasion in a dose- and time-dependent manner. The levels of LDH, IL-1ß and IL-18 of matrine combined with docetaxel-treated DU145 and PC-3 cells were significantly increased, compared with the untreated control cells. Flow cytometry, as well as Annexin-V/PI staining, showed a significant and dose-dependent increase in the number of early, as well as late-stage apoptotic cells in both DU145 and PC-3 cells compared with the untreated control cells. Western blot analysis showed that matrine combined with docetaxel treatment led to the expression of caspase1 in both DU145 and PC-3 cells. CONCLUSION: It may be more effective to use matrine in combination with docetaxel to treat androgen-resistant prostate cancer because matrine can help to affect proliferation, migration, invasion, apoptosis, metabolism, and have anti-inflammation effect on the tumor cells.

[Analysis of the phenotype-genotype relationship of hemoglobin Q-Thailand in Guangxi].

OBJECTIVE: To investigate the hematological and molecular characteristics of hemoglobin Q-Thailand in Guangxi, so as to provide reference data for hemoglobinopathy screening. METHODS: A total of 51088 samples were screened by capillary electrophoresis. Samples suspected with Hb Q-Thailand were processed with blood cell count and DNA sequencing. Gap-PCR and PCR-reverse dot blotting were used for the detection of common mutations of alpha and beta thalassemia. RESULTS: The carrier rate of Hb Q-Thailand in Guangxi was 0.06%. The hematological phenotype index(HGB, MCV, MCH, Hb Q-Thailand, Hb A2, Hb QA2) of 28 Hb Q-Thailand heterozygous samples were (125.60±22.30) g/L, (78.22±4.81) fl, (25.79±2.14) pg, (27.37±2.72)%, (1.89±0.22)%, (0.69±0.16)%, respectively, and of 2 Hb Q-Thailand heterozygous combined with beta-thalassemia samples were (125.00±18.39) g/L, (69.65±5.02) fl, (22.00±0.0) pg, (14.80±0.71)%, (4.45±0.07)%, (0.95±0.71)%, respectively. A statistical difference was found in hematological phenotype index between the two groups except HGB (P<0.05). CONCLUSION: In Guangxi, the detected Hb Q-Thailands were mainly heterozygous. Part of Hb Q-Thailand heterozygotes had normal red blood cell parameters, but can still be detected by hemoglobin electrophoresis. When combined with other types of thalassemia, these heterozygotes may still exhibit reduced MCV and MCH or various degrees of anemia.

The Establishment of Adult Reference Intervals on Fructosamine in Beijing.

BACKGROUND: Fructosamine (FRA) is widely used for diabetes monitor and control as a glycemic marker, especially in patients in whom the measurement of HbA1c may be biased or even unreliable. However, the FRA reference intervals based on Asian population features still keep seldom reported. The objective of this study was to establish the adult FRA reference intervals in Beijing, China. METHODS: A total of 1,497 healthy subjects were separated into three groups by gender and age. Subsequently, FRA levels in the collected serum samples from the reference individuals were tested by automatic chemical analyzer. The obtained data were statistically analyzed with SPSS. RESULTS: The serum FRA level in female group was slightly higher than that in male group without statistical significance. Meanwhile, further analysis indicated that the FRA level gradually increased along with the growth of the age. Compared with the age 20-45 group (248.83 ± 17.64 µmol/l) or the age 46-65 group (251.95 ± 19.63 µmol/l), the FRA level of the age >65 group (264.63± 23.05 µmol/l) was statistically significantly increased (P < 0.01). To better analyze the difference, the age 20-45 group and the age 46-65 group were combined into an age 20-65 group (249.88 ±18.39 µmol/l). In comparison to the age >65 group, the FRA level of age 20-65 group was significantly decreased (P <0.01). CONCLUSION: A novel FRA reference interval of the local healthy population in Beijing was established. The data demonstrated that there was no gender difference in FRA level, however, which was significantly increased in elder persons.

Prevalence and antimicrobial susceptibility of Ureaplasma urealyticum and Mycoplasma hominis in female outpatients, 2009-2013.

PURPOSE: The aim of this study was to estimate the prevalence and antimicrobial susceptibility of Ureaplasma urealyticum and Mycoplasma hominis among female outpatients treated for genital infection at a Chinese hospital from January 1, 2009 to December 31, 2013. METHODS: Samples from 6051 female outpatients were analyzed using Mycoplasma Identification and Antimicrobial Susceptibility Testing (ID/AST). RESULTS: The overall prevalence of U. urealyticum was higher than the prevalence of single M. hominis infection (31.2% vs 0.7%) and coinfections (31.2% vs. 1.9%). The percentage of U. urealyticum and/or M. hominis detected in the 30-39 year age group was greater than in the other age groups. More than 94.6% of the U. urealyticum isolates, 100% of the M. hominis isolates, and 84.3% of the isolates from coinfections were susceptible to doxycycline, minocycline, and tetracycline. More than 69.2% of the U. urealyticum isolates were susceptible to azithromycin, erythromycin, clarithromycin, and roxithromycin, but > 95.6% of the M. hominis isolates and 89.6% of the isolates from coinfections were resistant to these antibiotics. Acetylspiramycin, sparfloxacin, levofloxacin, ciprofloxacin, and ofloxacin were inactive against more than one-half of the isolates. More than 75.6% of the M. hominis isolates were susceptible to spectinomycin, but > 87.1% of the U. urealyticum and 93.3% of the coinfection isolates were resistant to this antibiotic. Isolates from three coinfections were completely resistant to the 14 antibiotics. CONCLUSION: The determination of antimicrobial susceptibility of these mycoplasma species is often crucial for optimal antimicrobial therapy of infected outpatients.

Urinary tract infection caused by Enterococcus isolates: aetiology and antimicrobial resistance patterns.

The purpose of this study was to analyse the data on resistance in Enterococcus faecalis and E. faecium isolated from urine samples of inpatients from January 2010 to December 2013. Compared to E. faecalis, E. faecium isolates were significantly more resistant to ampicilin, nitrofurantoin, and ciprofloxacin in the antimicrobial agents evaluated. Both species showed higher resistance to high-level gentamicin. The rate of vancomycin-resistant enterococci (VRE) in E. faecium was higher as compared to that of E. faecalis, and we found that apparent increase in the frequency of VRE E. faecalis clinical isolates in the 4-year study period. In our study, analysis of the antimicrobial resistance trends showed resistance to linezolid and vancomycin were <30.2% and <20.3% in both species, respectively. Of noteworthy is the high rate of MDR in two species from inpatients. This study highlights that it is of importance for clinicians to promote rational drug utilization and delay the emergence of resistant organisms.