The purpose of this study was to evaluate the application value of targeted next-generation sequencing(tNGS)for rapid detection of Mycobacterium tuberculosis.From September 2021 to October 2024,1 178 clinical samples from hospitalized pa-tients at the Non-tuberculous Mycobacteria Disease Diagnosis and Treatment Center of Guangzhou Chest Hospital were collected,in-cluding 272 sputum samples,871 broncho alveolar lavage fluid(BALF)samples,and 35 other samples.These samples were analyzed with tNGS and the liquid culture+DNA microarray chip method(referred to as the"culture identification method"),and the detection results between methods were compared.The numbers of Mycobacterium tuberculosis complex(MTBC)samples identified with tNGS and the culture identification method were 172 and 127,respectively,and the numbers of non-tuberculous mycobacteria(NTM)samples detected were 517 and 474,respectively.The detection rate of Mycobacterium was 56.88%with tNGS and was significantly lower(49.49%)with the culture identification method(χ2=13.27,P<0.05).For NTM identification,when sputum was used,the detection rate of tNGS was higher than that of the culture identification method(χ2=24.14,P<0.05).However,when BALF was used,no significant difference in detection rates was observed between tNGS and the culture identification method(χ2=3.97,P=0.06).When different sample types from the same patient were analyzed with tNGS for NTM,BALF was found to be a better choice than sputum(χ2=4.05,P<0.05).However,with the culture identification method,we observed no significant difference between sample types(χ2=2.72,P=0.10).Furthermore,both the MTBC and NTM sequences detected with tNGS were significantly higher in the culture-positive group than the culture-negative group.With increasing sequence numbers,the proportion of cases in the culture-positive group progressively rose,whereas an inverse trend was observed in the culture-negative group.With clinical diagno-sis as the reference standard,the sensitivity,specificity,and Kappa value of tNGS and the culture identification method in identifying mycobacterial diseases were 85.41%,91.32%,0.74 and 74.73%,93.15%,0.63,respectively.Compared with the culture identifica-tion method,tNGS for rapid detection of Mycobacterium exhibited excellent sensitivity,specificity and consistency,and its timeliness should help clinicians make precise individualized treatment plans earlier.

The purpose of this study was to evaluate the application value of targeted next-generation sequencing(tNGS)for rapid detection of Mycobacterium tuberculosis.From September 2021 to October 2024,1 178 clinical samples from hospitalized pa-tients at the Non-tuberculous Mycobacteria Disease Diagnosis and Treatment Center of Guangzhou Chest Hospital were collected,in-cluding 272 sputum samples,871 broncho alveolar lavage fluid(BALF)samples,and 35 other samples.These samples were analyzed with tNGS and the liquid culture+DNA microarray chip method(referred to as the"culture identification method"),and the detection results between methods were compared.The numbers of Mycobacterium tuberculosis complex(MTBC)samples identified with tNGS and the culture identification method were 172 and 127,respectively,and the numbers of non-tuberculous mycobacteria(NTM)samples detected were 517 and 474,respectively.The detection rate of Mycobacterium was 56.88%with tNGS and was significantly lower(49.49%)with the culture identification method(χ2=13.27,P<0.05).For NTM identification,when sputum was used,the detection rate of tNGS was higher than that of the culture identification method(χ2=24.14,P<0.05).However,when BALF was used,no significant difference in detection rates was observed between tNGS and the culture identification method(χ2=3.97,P=0.06).When different sample types from the same patient were analyzed with tNGS for NTM,BALF was found to be a better choice than sputum(χ2=4.05,P<0.05).However,with the culture identification method,we observed no significant difference between sample types(χ2=2.72,P=0.10).Furthermore,both the MTBC and NTM sequences detected with tNGS were significantly higher in the culture-positive group than the culture-negative group.With increasing sequence numbers,the proportion of cases in the culture-positive group progressively rose,whereas an inverse trend was observed in the culture-negative group.With clinical diagno-sis as the reference standard,the sensitivity,specificity,and Kappa value of tNGS and the culture identification method in identifying mycobacterial diseases were 85.41%,91.32%,0.74 and 74.73%,93.15%,0.63,respectively.Compared with the culture identifica-tion method,tNGS for rapid detection of Mycobacterium exhibited excellent sensitivity,specificity and consistency,and its timeliness should help clinicians make precise individualized treatment plans earlier.

Laminin(LN) and hyaluronie acid (HA) are extracellular matrix in liver. The effects of LN and HA on human fetal liver cells proliferation and collagen synthesis were observed and serum levels of LN and HA were assayed m patients with liver diseases by an enzyme immunoassay method. Both LN and HA had a significant effect on the inhibition of cell proliferation and collagen production. However, serum levels of LN and HA were significantly increased in liver cirrhosis comparing with healthy controls and with patients with non-liver diseases. The results suggest that LN and HA may play a role in the feed-back inhibition mechanism in liver fibrogencsis, and assay of LN and HA in serum may be useful to the diagnosis of liver fibrosis.

The glucocorticoid receptor of hepatic cytosol of rat was determined by the exchange assay in the presence of endogenous corticosterone. The condition of ex-chame was 0℃ , 4.5~5 hours. The dissociation of corticosterone-receptor complex was about 90% under such a condition. The effect of the endogenous corticosterore on the assay was evaluated by the measurement of [3H]-dexamethasone binding after preincubation of the liver cytosol from adrenalectomized rats with different amounts of corticosterone. The results indicated that, at a corticosterone concentration of less than 6 nM the error of the specific binding capacity (Ro) of [3H]-dexa-methasone was no more than 10%,and the apparant dissociation constant (Kd) was essentially unchanged. The Ro and Kd of glucocorticoid receptor of the hepatic cytosol of normal and scalded rats were measured by this method. The Ro was decressed in the scalded rats. The levels of Ro in hepatic cytosol from rats after 1, 12,24, 36 hours and 5 days of scalding were 76.4?15.2%,70.6?22.0%79.7?5.8%, 113.0?24.8% and 102.9 ?17.6% (mean?SD) of the control group respectively. The apparant Kd slightly increased 1 hour after scalding. The possible mechanisms of the above changes were discussed.

The high-affinity glucocorticoid binding sites (HAGS) and the low-affinity glucocorticoid binding sites (LAGS) with steroid specificity were demostrated in cerebral cytosol of rats by using the radioligand binding assay. The Kd of HAGS and LAGS were (178?0.71)?l0-8mol/L and (2.12?1.06)?10-6mol/L respecitively as estimated by Scatchard and Pseudoscatchard analysis. Glucocorticoid receptors (GR) in the traumatized(left) hemisphere cytosol were decreased more significantly than those in both the control (right) hemisphere cytosol at 6h postinjury and normal brain tissue (P