We performed the transforming nitrate test mainly with penicillium brevi-compactum and other fungi. The strain of P. brevi-compactum and all the other fungi used for this test were isolated from patients with cervix uteri carcinoma, patients with gynopathy but no cancer, and the normal woman controls in Luoyang county of Shaanxi and in Wufeng county of Hubei respectively, which are the highest prevalance areas of cervix uteri carcinoma in China. The detection rate of P. brevicompactum is higher 72.73% (65/77) in Luoyang; 67.65 (23/34) in Wufeng. The detection rate of yeast and Candida is low. Though the detection rate of certain tested fungi such as A. serreus, A. candidus, P. cyelopium and Rhodotorula rubra is higher, these fungi are seldom found in the isolates from cervix uteri. Thus, these fungi might be no important significance in etiology of cervix uteri carcinoma. Because nitriamine is an important carcinogen and the effect of transforming nitrate to nitrite plays a more important role in the synthesis of nitriamine, P. brevi-compactum is the dominant strain of fungi isolated from the patient with cervix uteri carcinoma in this investigation. The higher detection rate of the treated P. brevi-compactum shows that this fungus plays a role in the etiology or incidence of cervis uteri carcinoma. To solve this difficult and complex problem of the role that fungal agent okay in the etiology of carcinoma further studies must be done.

Objective To investigate the correlation between quantitative parameters with contrastenhanced ultrasound (CEUS) and microvessel density (MVD).Methods Thirty-four placenta of rat model of preeclampsia underwent CEUS examination.The peak intensity time curves on the enhanced images were analyzed quantitatively with computer to get quantitative parameters[the time to peak(TTP), maximal peak intensity(Imax), the area under curve(AUC) and the mean perfusion volume(V)].These parameters were compared with MVD counted with immunohistochemistry and the correlation was statistically studied.Results The TTP in the enhanced images was (14.55 ± 3.45)s, Imax was (20.83 ± 6.15) dB, AUC was (1868.61 ± 25.76)dB, V was (58.01 ± 23.56)dB, and the MVD of placenta of rat model of preeclampsia was (88.98 ± 24.78) in 34 rats.The Imax was correlated positively to MVD (r = 0.885, P = 0.000) ,AUC was correlated positively to MVD (r = 0.677, P = 0.001), V was correlated positively to MVD (r =0.877, P = 0.000).There was no correlation between TTP and MVD in lesions.Conclusions The Imax,AUC and V calculated with CEUS were correlated to MVD, these parameters were valuable index for quantitative evaluation of placental blood perfusion.

Objective: To analyze the association of cytogenetic abnormalities with the prognosis of chronic myeloid leukemia (CML) patients in tyrosine kinase inhibitors (TKI) era. Methods: Karyotype analysis of chromosome G-banding was carried out in 387 newly diagnosed CML patients by short-term culture of bone marrow cells. The correlation of cytogenetic abnormalities and CML progression was explored in combination with ABL tyrosine point mutations. Result: Of 387 patients with positive BCR-ABL fusion gene assayed by fluorescence in situ hybridization (FISH) technique, 94.1% (364/387) patients were Ph positive and 5.9% (23/387) Ph negative; 320 patients (87.9%) had a translocation t (9;22) (q34;q11) and 5 (1.4%) a variant translocation t (v;22) . Additional cytogenetic aberrations (ACA) at diagnosis were found in 10.7% (39/387) Ph⁺ patients, major route ACA in 22 (56.4%) cases and minor route ACA in 15 (38.5%) cases and 2 patients (5.1%) lacked the Y chromosome (−Y) ; 23.4% (71/303) patients occurred ACA during TKI treatment and the most frequent abnormalities were abnormal chromosome numbersd, which were likely associated with high proportion of disease progression (χ²=168.21, P<0.001) and ABL tyrosine point mutations (χ²=29.04, P<0.001) . Newly diagnosed CML-CP patients with t (9;22) (q34;q11) had a longer event-free survival (EFS) and disease-free survival (DFS) rates than that of patients with ACA (P=0.037; P=0.003) , while the overall survival (OS) had no significant differences (P=0.209) . As for CML-CP patients that occurred ACA during TKI therapy would have a marked low OS, EFS and DFS (all P<0.001) compared with no ACA occurred patients. Survival of advanced patients that occurred ACA were dramatically reduced. Conclusion ACA often emerged during the disease progress in CML patients, regular and timely detection of chromosomes karyotype and ABL tyrosine point mutations during TKI treatment was important for therapeutic evaluation, progress and prognosis of CML.

Objective: To investigate the clinical implications of p16 gene deletion in adult Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph⁺ ALL) . Methods: Retrospective analysis of clinical, immunophenotypic, cytogenetics, molecular characteristics and prognosis of 80 newly diagnosed Ph⁺ ALL patients with p16 deletion. Results: Of 80 adult Ph⁺ ALL, the prevalence of p16 gene deletion was 31.3%. p16 gene deletion carriers frequently accompanied with high WBC counts (WBC≥30×10⁹/L) and CD20 expression. The incidence of complex chromosome abnormality in p16 gene deletion group was higher than that in non-deletion group, with alternations in chromosome 7, 8, 19 and der (22) more frequently observed. There was no difference occurred between patients with or without p16 gene deletion in complete remission (CR) rate following induction chemotherapy combined with tyrosine kinase inhibitors (TKIs) . However, after three cycles of chemotherapy, the MMR and CMR rate in the p16 gene deletion group was lower than patients with wild-type p16 gene (P=0.034, P=0.036) . The p16 gene deletion patients showed no significant differences in MMR, CMR and relapse rate between Imatinib or Dasatinib plus chemotherapy (P>0.05) . Deletion of p16 gene was significantly associated with poor outcomes including worse overall survival (OS) (37.1% vs 54.1%, P=0.037) , lower disease free-survival (DFS) (12.4% vs 45.9%, P=0.026) , and increased cumulative incidence of relapse (P=0.033) . Among the 25 patients with p16 deletion, 14 underwent allo-HSCT and the median survival was 21 months, better than that of patients received chemotherapy alone (12 months) (P=0.030) . Conclusion This study indicated that deletion of p16 was associated with poor prognosis in adult Ph⁺ ALL, and the utility of second-generation TKI (Dasatinib) does not necessarily have an edge on efficacy over Imatinib, but allo-HSCT has the potential of elongating life expectancy. It is an important significance to define the status of p16 in Ph⁺ ALL for predicting prognosis and guiding therapy decision-making.

Objective To investigate the quantitative evaluation efficiency of gadolinium?ethoxybenzyl?diethylenetriamine pentaacetic acid (Gd?EOB?DTPA) enhanced T1 mapping in staging hepatic fibrosis caused by viral hepatitis B. Methods One hundred and fifty patients with chronic hepatitis B were prospectively enrolled in Shanghai Public Health Clinical Center, Fudan University from August 2016 to August 2018.These patients underwent liver aspiration biopsy were divided into four subgroups: S1 (n=38), S2 (n=30), S3 (n=33), S4 (n=49) according to Scheuer?Ludwig scoring system. Non?enhanced and Gd?EOB?DTPA?enhanced MRI were performed in all subjects. Look?Locker sequences were performed to acquire T1 mapping of pre and post?contrast at 20 minutes after Gd?EOB?DTPA administration. The T1 value after 20 minutes of Gd?EOB?DTPA administration (T1 20 min), the reduction rate of T1 value (ΔT1 20 min% ), the increase of 1/T1 value (ΔR1 20 min% ) were measured and calculated. The one?way ANOVA was applied to compare the difference in T1 20 min, ΔT1 20 min%, ΔR1 20 min% of various fibrosis stages. ROC curves were used to assess the efficacy of T1 20 min, ΔT1 20 min%, ΔR1 20 min% for diagnosing≥S2,≥S3,≥S4. P<0.05 was considered to be statistically significant. Results The T1 20 min raised with fibrosis stage increased, ΔT1 20 min% and ΔR1 20 min% reduced with fibrosis stage increased. Areas under the curves of T1 20 min, ΔT1 20 min%, ΔR1 20 min% for diagnosing≥S2 were 0.844, 0.905, 0.869; and diagnosing≥S3 were 0.832, 0.907, 0.862; and diagnosing≥S4 were 0.853, 0.897, 0.873, respectively. The diagnostic efficiency of ΔT1 20 min% was the best. Conclusion Gd?EOB?DTPA?enhanced T1 mapping could be regarded as a reliable diagnostic tool for the evaluation of hepatic fibrosis caused by viral hepatitis B.

In this study, we attempted to establish a culture system for in vitro spermatogenesis from spermatogonial stem cells (SSCs) of Bama mini-pig. Dissociated testicular cells from 1-month-old pigs were co-cultured to mimic in vivo spermatogenesis. The testicular cells were seeded in minimum essential medium alpha (α-MEM) supplemented with Knockout serum replacement (KSR). Three-dimensional colonies formed after 10 days of culture. The colonies showed positive staining for SSC-associated markers such as UCHL1, PLZF, THY1, OCT4, Dolichos biflorus agglutinin, and alkaline phosphatase. Induction of SSCs was performed in α-MEM + KSR supplemented with retinoic acid, bone morphogenetic protein 4, activin A, follicle-stimulating hormone, or testosterone. The results showed that STRA8, DMC1, PRM1, and TNP1 were upregulated significantly in the colonies after induction compared to that in testis from 1-month-old pigs, while expression levels of those genes were significantly low compared to those in 2-month-old testis. However, upregulation of ACROSIN was not significant. Replacement of α-MEM and KSR with Iscove's modified Dulbecco's medium and fetal bovine serum did not upregulate expression of these genes significantly. These results indicate that SSCs of Bama mini-pig could undergo differentiation and develop to a post-meiotic stage in α-MEM supplemented with KSR and induction factors.
Acrosin ; Activins ; Alkaline Phosphatase ; Bone Morphogenetic Protein 4 ; Dolichos ; Follicle Stimulating Hormone ; Humans ; In Vitro Techniques ; Infant ; Infant, Newborn ; Spermatogenesis ; Stem Cells ; Swine ; Testis ; Testosterone ; Tretinoin ; Up-Regulation

Objective: To investigate the quantitative evaluation efficiency of gadolinium- ethoxybenzyl-diethylenetriamine pentaacetic acid (Gd-EOB-DTPA) enhanced T₁ mapping in staging hepatic fibrosis caused by viral hepatitis B. Methods: One hundred and fifty patients with chronic hepatitis B were prospectively enrolled in Shanghai Public Health Clinical Center, Fudan University from August 2016 to August 2018.These patients underwent liver aspiration biopsy were divided into four subgroups: S1 (n=38), S2 (n=30), S3 (n=33), S4 (n=49) according to Scheuer-Ludwig scoring system. Non-enhanced and Gd-EOB-DTPA-enhanced MRI were performed in all subjects. Look-Locker sequences were performed to acquire T₁ mapping of pre and post-contrast at 20 minutes after Gd-EOB-DTPA administration. The T₁ value after 20 minutes of Gd-EOB-DTPA administration (T_{1 20 min}), the reduction rate of T₁ value (ΔT_{1 20 min}%), the increase of 1/T₁ value (ΔR_{1 20 min}%) were measured and calculated. The one-way ANOVA was applied to compare the difference in T_{1 20 min}, ΔT_{1 20 min}%, ΔR_{1 20 min}% of various fibrosis stages. ROC curves were used to assess the efficacy of T_{1 20 min}, ΔT_{1 20 min}%, ΔR_{1 20 min}% for diagnosing ≥ S2, ≥ S3, ≥ S4. P<0.05 was considered to be statistically significant. Results: The T_{1 20 min} raised with fibrosis stage increased, ΔT_{1 20 min}% and ΔR_{1 20 min}% reduced with fibrosis stage increased. Areas under the curves of T_{1 20 min}, ΔT_{1 20 min}%, ΔR_{1 20 min}% for diagnosing ≥ S2 were 0.844, 0.905, 0.869; and diagnosing ≥ S3 were 0.832, 0.907, 0.862; and diagnosing ≥ S4 were 0.853, 0.897, 0.873, respectively. The diagnostic efficiency of ΔT_{1 20 min}% was the best. Conclusion Gd-EOB-DTPA-enhanced T₁ mapping could be regarded as a reliable diagnostic tool for the evaluation of hepatic fibrosis caused by viral hepatitis B.

ObjectiveBased on the experience of traditional quality evaluation， the quality of Atractylodis Macrocephalae Rhizoma（AMR） with different production methods such as direct seeding， transplanting after seedling raising， topping and non-topping， and difference in growth years was compared. MethodVernier caliper was used to measure the trait data of AMR in different production methods. Paraffin sections of AMR with different production methods were made by saffron solid green staining， and the microstructure was observed. The contents of water-soluble and alcohol-soluble extracts in AMR with different production methods were determined according to the 2020 edition of Chinese Pharmacopoeia. The content of water-soluble total polysaccharides in AMR with different production methods was detected by sulfuric acid-anthrone method. Fiber analyzer was used to detect the content of fiber components in AMR with different production methods. The contents of monosaccharides， oligosaccharides and some secondary metabolites in AMR with different production methods were detected by ultra performance liquid chromatography（UPLC）， and the differences of chemical components were compared by multivariate statistical analysis methods such as principal component analysis（PCA） and partial least squares-discriminant analysis（PLS-DA）. ResultIn terms of traits， the 3-year-old AMR with direct seeding and without topping was close to the high-quality AMR with "phoenix-head and crane-neck， strong sweetness and clear aroma" recorded in ancient materia medica， followed by the 3-year-old AMR with topping after transplanting， while the 2-year-old AMR with topping after transplanting with high market circulation rate was generally fat and strong with mild odor. In the microscopic aspect， the arrangement of xylem vessels and fiber bundles in the 3-year-old samples formed two obvious rings. Compared with the 2-year-old samples cultivated in Bozhou and Zhejiang， the 3-year-old samples without topping after transplanting had more wood fibers. In terms of chemical composition， the contents of 70% ethanol extract， fructose， glucose， sucrose， 1-kestose， atractylenolide Ⅰ， chlorogenic acid， neochlorogenic acid， cryptochlorogenic acid and other components in 3-year-old AMR with direct seeding and without topping were significantly higher than those in the other three samples（P<0.05）. The contents of cellulose， 70% ethanol extract， sucrose， atractylenolide Ⅰ， atractylone and other components in 3-year-old AMR with topping after transplanting were significantly higher than those in the 2-year-old AMR with high market circulation rate（P<0.05）， while the contents of water-soluble extract and water-soluble total polysaccharides in 2-year-old samples with topping after transplanting were significantly higher than those in the 3-year-old AMR with topping after transplanting， direct seeding and without topping（P<0.05）. ConclusionUnder the current mainstream production mode， too much manual intervention makes AMR heavily enriched in polysaccharides and increased the yield， but the accumulation of sweet substances， fragrant substances and fiber substances is insufficient， which affects its quality. The current quality standard of AMR has some shortcomings in guiding the high quality production of it， it is suggested to revise the quality standard of AMR， supplement the quantitative analysis of secondary metabolites， and strengthen the production of imitation wild AMR.