Representative images of immunohistochemical staining of inducible heat shock protein 70 (HSP70) in pellet culture samples on (A) Day 17 (B) Day 24.

<p>Scale bar: 25 μm. (chon: chondrogenic differentiated hMSCs, and chon+HS: chondrogenic differentiated hMSCs with heat shock).</p

Representative images of immunohistochemical staining of aggrecan in pellet culture samples on (A) Day 17 (B) Day 24.

<p>Scale bar: 25 μm. (chon: chondrogenic differentiated hMSCs, and chon+HS: chondrogenic differentiated hMSCs with heat shock).</p

Sulfated GAG (sGAG) content normalized to wet weight in 3D pellet culture on Day 10, 17 and 24, with and without HS (chon: chondrogenic differentiated hMSCs, and chon+HS: chondrogenic differentiated hMSCs with heat shock).

<p>Data represent the mean ± SD (n = 4). P values were calculated using student t-test.</p

Semi-quantified Immunohistochemical staining Intensity.

<p>Chon: Chondrogenic; HS: Heat Shock; Col: Collagen; n = 3 for average ratio.</p

Characterization of hMSCs by flow cytometric analysis.

<p>(<b>A</b>) Isolated hMSCs were positive for surface markers CD146, CD29, CD147 and CD44, and negative for CD45 and CD34. (<b>B</b>) The individual percentages of each surface marker expressed in these cells from the quantitative FACS analysis. Data represent the mean ± SD (n = 3).</p

Western Blot analysis of collagen type II, aggrecan, and HSP70 expression in 3D chondrogenic pellet cultures using hMSCs from the 24 year old donor at Day 17 and Day 24.

<p>(A), (C), (E), (G), (I) and (K) are the images of Western blot membranes while (B), (D), (F), (H), (J) and (L) are their semi-quantified band intensities respectively (Chon: chondrogenic differentiated hMSCs, and chon+HS: chondrogenic differentiated hMSCs with heat shock).</p

Table_1_The incremental value of Mycobacterium tuberculosis trace nucleic acid detection in CT-guided percutaneous biopsy needle rinse solutions for the diagnosis of tuberculosis.DOCX

IntroductionTuberculosis (TB) diagnosis still faces challenges with high proportion of bacteriologic test negative incidences worldwide. We assessed the diagnostic value of digital PCR (dPCR) analysis of ultramicro Mycobacterium tuberculosis (M.tb) nucleic acid in CT-guided percutaneous biopsy needle rinse solution (BNRS) for TB.MethodsBNRS specimens were consecutively collected and total DNA was purified. The concentrations of M.tb-specific IS6110 and IS1081 were quantified using droplet dPCR. The diagnostic performances of BNRS-dPCR and its sensitivity in comparison with conventional tests were analyzed.ResultsA total of 106 patients were enrolled, 63 of whom were TB (48 definite and 15 clinically suspected TB) and 43 were non-TB. The sensitivity of BNRS IS6110 OR IS1081-dPCR for total, confirmed and clinically suspected TB was 66.7%, 68.8% and 60.0%, respectively, with a specificity of 97.7%. Its sensitivity was higher than that of conventional etiological tests, including smear microscopy, mycobacterial culture and Xpert using sputum and BALF samples. The positive detection rate in TB patients increased from 39.3% for biopsy AFB test alone to 73.2% when combined with BNRS-dPCR, and from 71.4% for biopsy M.tb molecular detection alone to 85.7% when combined with BNRS-dPCR.ConclusionOur results preliminarily indicated that BNRS IS6110 OR IS1081-dPCR is a feasible etiological test, which has the potential to be used as a supplementary method to augment the diagnostic yield of biopsy and improve TB diagnosis.</p