Tumor cell-based glycolytic metabolism and single-cell sequencing of urinary exfoliated cells for the diagnosis and molecular profiling of urothelial carcinoma / 中华病理学杂志

Objective: To explore the diagnostic values of HK2 testing and single-cell sequencing in the urothelial carcinoma (UC). Methods: The qualified urine specimens of 265 suspected UC patients or postoperative patients from the Cancer Hospital of Chinese Academy of Medical Sciences, Beijing, China were collected. Both exfoliative cytology and HK2 testing were performed on clinically suspected UC or postoperative patients. The performance of diagnostic cytology and HK2, including consistency, sensitivity, specificity, positive predictive value and negative predictive value, was evaluated based on histopathological, clinical and imaging diagnosis. Isolated HK2 metabolically abnormal cells were subject to single-cell sequencing to verify the reliability of HK2 detection performance and to explore the molecular characteristics of UC. Results: The concordance rate of HK2 testing and cytology for detecting UC was 90.3% (102/113, Kappa=0.604). Compared with cytology, the sensitivity of HK2 was significantly higher (85.2% versus 75.6%, P=0.024). The detection sensitivity of combined HK2 testing and cytology was increased to 91.1%. HK2 testing was significantly more sensitive than cytology for diagnosing UC in the upper urinary tract (81.8% versus 65.5%, P=0.022). It was also more sensitive than cytology for diagnosing early-stage UC (82.6% versus 69.5%, P=0.375) and low-grade UC (69.6% versus 47.8%, P=0.125). Single-cell sequencing of the ten patients, whose samples were positive for HK2, demonstrated highly concordant copy number variations (CNVs) in tumor cells from the same UC patient, with heterogeneity in CNV profiles among different patients. Deletion of chromosome 8p was found in 3 of the 4 urine samples of renal pelvis UC. The 2 patients with benign lesions had no CNVs in all sequenced cells. Conclusions: The test for abnormal urinary glycolytic HK2 metabolism can assist urine cytology to improve the sensitivity of UC diagnosis, and it provides a novel and reliable approach for early detection of upper urinary tract UC and lower grade UC. Meanwhile, this study has preliminarily revealed the feasibility of single-cell sequencing in urinary samples, which is expected to improve the diagnostic specificity of HK2 testing.

Research progress of polymer self-healing mechanism and its application in biomedicine / 药学学报

Polymer self-healing is mainly based on the molecular structure and interaction of polymers, and some need external stimulation, such as light, heat, pH, etc. In recent years, many studies have found that the self-healing properties of polymers can prolong the life of materials, while maintaining the mechanical properties of polymers after healing. According to the different action modes of polymer materials, it can be divided into autonomous self-healing and non-autonomous self-healing. Among them, autonomous self-healing mainly works through reversible covalent bonds (Schiff base bond, Diels-Alder reaction, hydrazide bond), reversible non-covalent bonds (hydrogen bond, metal-ligand coordination bond, electrostatic interaction, π-π stacking interaction, hydrophobic interaction) and a combination of the two interactions. Drug carriers with unique self-healing properties play an important role in the encapsulation and stable release of biomacromolecules. In this review, the self-healing mechanism of polymers and their applications in the field of biomedicine were briefly summarized and discussed.

Puerarin Inhibits the Proliferation,Invasion,and Migration of Non-small Cell Lung Cancer Cells through Regulating miR-490/Denticleless E3 Ubiquitin Protein Ligase / 中国医学科学院学报

Objective To explore the mechanism of puerarin inhibiting the proliferation,invasion,and migration of non-small cell lung cancer cells. Methods A549 cells were cultured and treated with different concentrations of puerarin.The inhibition rate (IR) on cell proliferation was detected by CCK-8,and qRT-PCR was performed to detect the mRNA levels of miR-490 and denticleless E3 ubiquitin protein ligase(DTL).Double luciferase reporter assay was employed to identify the targets of miR-490 and DTL based on the establishment of NC mimic group,miR-490 mimic group,NC inhibitor group,and miR-490 inhibitor group.The cells treated by 20 μmol/L puerarin were classified into six groups:DMSO,puerarin,puerarin+NC inhibitor,puerarin+miR-490 inhibitor,puerarin+miR-490 inhibitor+Si-NC,and puerarin+miR-490 inhibitor+Si-DTL.Transwell was used to detect cell migration and invasion.Western blotting was performed to detect the protein levels of epithelial-mesenchymal transition-related markers E-cadherin,N-cadherin,and Vimentin. Results With the increase in puerarin concentration,the IR gradually elevated (F=105.375,P<0.001),miR-490 expression gradually increased (F=32.919,P<0.001),and DTL expression gradually decreased (F=116.120,P<0.001).Compared with NC mimic group,miR-490 mimic group had decreased luciferase activity (t=7.762,P=0.016),raised miR-490 mRNA level (t=13.319,P<0.001),and declined DTL mRNA level (t=7.415,P=0.002).Compared with those in NC inhibitor group,miR-490 demonstrated decreased mRNA level (t=9.523,P=0.001) and DTL presented increased mRNA level (t=11.305,P<0.001) in miR-490 inhibitor group.Western blotting showed that the protein level of DTL was higher in NC mimic group (t=7.953,P=0.001) than in miR-490 mimic group and higher in miR-490 inhibitor group than in NC inhibitor group (t=10.552,P<0.001).Compared with DMSO group,puerarin group showed up-regulated mRNA level of miR-490 (t=10.255,P=0.001) while down-regulated mRNA level of DTL (t=6.682,P=0.003).Compared with those in puerarin+NC inhibitor group,the mRNA level of miR-490 declined (t=10.995,P<0.001) while that of DTL raised (t=12.478,P<0.001) in puerarin+miR-490 inhibitor group.The mRNA level of miR-490 had no significant difference between puerarin+miR-490 inhibitor+Si-NC group and puerarin+miR-490 inhibitor+Si-DTL group (t=1.081,P=0.341),and that of DTL was lower in the latter group (t=14.321,P<0.001).The protein level of DTL was higher in puerarin+miR-490 inhibitor group than in puerarin+NC inhibitor group (t=11.423,P<0.001),and lower in puerarin+miR-490 inhibitor+Si-DTL group than in puerarin+miR-490 inhibitor+Si-NC group (t=12.080,P<0.001).Compared with DMSO group,puerarin group showed inhibited cell proliferation (F=129.27,P<0.001).The activity of cell proliferation was higher in puerarin+miR-490 inhibitor group than in puerarin+NC inhibitor group (F=75.12,P<0.001),and higher in puerarin+miR-490 inhibitor+Si-NC group than in puerarin+miR-490 inhibitor+Si-DTL group (F=52.59,P<0.001).Compared with DMSO group,puerarin group had suppressed cell migration (t=8.963,P=0.001).The cell migration ability was higher in puerarin+miR-490 inhibitor group than in puerarin+NC inhibitor group (t=12.117,P<0.001) and higher in puerarin+miR-490 inhibitor+Si-NC group than in puerarin+miR-490 inhibitor+Si-DTL group (t=12.934,P<0.001).Puerarin group showed weakened cell invasion ability compared with DMSO group (t=4.710,P=0.009).The cell invasion ability was higher in puerarin+miR-490 inhibitor group than in puerarin+NC inhibitor group (t=13.264,P<0.001) and lower in puerarin+miR-490 inhibitor+Si-DTL group than in puerarin+miR-490 inhibitor+Si-NC group (t=13.476,P<0.001).Compared with DMSO group,puerarin group showed up-regulated protein level of E-cadherin (t=7.137,P=0.002) while down-regulated protein levels of N-cadherin (t=8.828,P=0.001) and vimentin (t=6.594,P=0.003).Compared with those in puerarin+NC inhibitor group,the protein level of E-cadherin (t=12.376,P<0.001) decreased while those of N-cadherin (t=13.436,P<0.001) and vimentin (t=11.467,P<0.001) increased in puerarin+miR-490 inhibitor group.Compared with puerarin+miR-490 inhibitor+Si-NC group,puerarin+miR-490 inhibitor+Si-DTL group up-regulated the protein level of E-cadherin (t=13.081,P<0.001) while down-regulated the protein levels of N-cadherin (t=10.835,P<0.001) and vimentin (t=11.862,P<0.001). Conclusion Puerarin could inhibit the proliferation,invasion,and migration of non-small cell lung cancer cells by up-regulating miR-490 and down-regulating DTL.

Ginkgo biloba extract ameliorates streptozotocin-induced diabetes in rats as measured by non-targeted metabolomics / 药学学报

Metabolomics based on liquid chromatography coupled with mass spectrometry (LC-MS) was used to study the initiation and development of diabetes in rats, and the ability of Ginkgo biloba extract (GBE) to ameliorate this pathology. Diabetes mellitus (DM) was induced by intra-peritoneal injection of streptozotocin. The rats were randomly divided into a normal control group treated with drug-free solution (NC), a normal control group treated with GBE (N-GBE), a DM group treated with drug-free solution (DM), and a DM group treated with GBE (D-GBE); rats were maintained on this protocol for 9 weeks. Rat plasma was collected from the sixth week to the ninth week and then analyzed with LC-MS. Animal experimentation was approved by the Committee on the Ethics of Animal Experiments of Xuzhou Medical University. Twelve plasma metabolites with continuous differentiation were monitored to indicate dysfunction of metabolic pathways including fatty acid metabolism, phospholipid metabolism, amino acid metabolism, tricarboxylic acid cycle activity, bile acid metabolism, and purine metabolism to confirm the occurrence and development of DM. Treatment with GBE partially reversed the changes seen in five metabolites in DM rats, indicating that GBE could prevent the occurrence and development of DM by acting on fatty acid metabolism, phospholipid metabolism, amino acid metabolism, and the tricarboxylic acid cycle.

Relationship between Electrical Conductivity and Chemical Content of Rat Skeletal Muscle Impregnating Solution and Postmortem Interval / 法医学杂志

Objective To study the mechanism of change of the electrical conductivity （EC） of rat skeletal muscle impregnating solution that occurs with the change of postmortem interval （PMI）. Methods Healthy Sprague-Dawley rats were killed and kept at about 25 ℃. Skeletal muscles were extracted at different PMI--immediate （0 d）, 1 d, 2 d, 3 d, 4 d, 5 d, 6 d, and 7 d, then mixed with deionized water to make impregnating solution with a mass concentration of 0.1 g/mL. The solution's EC and nine common chemicals in it, such as potassium ion, calcium ion, and chloride ion, were determined. Results EC increased gradually with the extending of PMI （P=0.024） during the 7 days after the rats' death. The content of uric acid （P=0.032）, urea nitrogen （P=0.013） and phosphorus （P=0.022） also increased during the extension. However, the content of magnesium ions decreased with extending of PMI （P=0.047）. The correlation between potassium ion, sodium ion, chlorine ion, calcium ion, creatinine and PMI were weak （P>0.05）. Conclusion The molecular basis of skeletal muscle EC change in rats after their death is the changes of uric acid, urea nitrogen, inorganic phosphorus and other chemical components. Furthermore, combine use of various indicators can improve the accuracy of the EC method to infer PMI.

Relationship between Electrical Conductivity and Related Biochemical Indicators of Rat Cerebrum Tissues and Postmortem Interval / 法医学杂志

Objective To explore the relationship between the electrical conductivity （EC） and biochemical indicators of rat cerebrum tissues and postmortem intervals （PMIs） and discuss the mechanism of applying EC to infer PMI. Methods Forty healthy Sprague-Dawley rats were sacrificed and stored in an environment of about 25 ℃. The whole cerebrum tissues of rats were removed respectively at different PMIs of 0, 1, 2, 3, 4, 5, 6, and 7 d, and then made into homogenized impregnation solution. The EC and related biochemical indicators （potassium, sodium, chloride, calcium, inorganic phosphorus, magnesium, uric acid, urea nitrogen and creatinine） in cerebrum tissue impregnation solution were determined, and the relationships among EC in impregnation solution, related biochemical indicators and PMI were analyzed. Results The EC in cerebrum tissues increased gradually with the extension of PMI, and the content of uric acid, urea nitrogen and inorganic phosphorus in its impregnation solution also increased gradually with the extension of PMI. The correlation of EC, uric acid, urea nitrogen, and inorganic phosphorus with PMI was relatively good （R2 was 0.95-0.99）, and there was a linear correlation between the content change of uric acid, urea nitrogen, inorganic phosphorus and EC （R2 was 0.97-0.99）. The changes of the other 6 kinds of biochemical indicators with the extension of PMI within 7 d after the rats' death were non-significant （P>0.05）. Conclusion The correlation between EC in cerebrum tissues, uric acid, urea nitrogen, inorganic phosphorus and PMI were relatively good, and combining various indicators can also improve the accuracy of PMI estimation.

Analysis of 28S rRNA and COⅠ Gene Sequence of Nine Necrophagous Calliphorid Flies from Luoyang / 法医学杂志

Objective To assess the feasibility of using 28S ribosomal RNA （28S rRNA） and mitochondrial cytochrome c oxidase subunit Ⅰ （COⅠ） gene sequences of nine necrophagous Calliphorid flies for the identification of common necrophagous Calliphorid flies, and to provide technical support for postmortem interval （PMI） estimation. Methods Twenty-three Calliphorid flies were collected and identified morphologically, and DNA were extracted from legs. The gene fragments of 28S rRNA and COⅠ were amplified and sequenced, then the sequence alignment was performed with BLAST. The composition of obtained sequences was analyzed and evolutionary divergence rate between species and intraspecies were established. The phylogeny tree was constructed with neighbor-joining method. Results The 23 necrophagous Calliphorid flies were identified to 9 species of 5 genera. The 715 bp from 28S rRNA and 637 bp from COⅠ gene were obtained and the online BLAST result showed more than 99% of similarity. The phylogeny tree showed that the necrophagous flies could cluster well into 9 groups, which was consistent with morphological identification results. The intraspecific difference in 28S rRNA was 0 and the interspecific difference was 0.001-0.033. The intraspecific difference in COⅠ was 0-0.008 and the interspecific difference was 0.006-0.101. Conclusion Combined use of 28S rRNA and COⅠ gene sequence fragments can effectively identify the nine Calliphorid flies in this study. However, for closely related blowfly species, more genetic markers should be explored and used in combination in future.

Molecular Identification of Sarcosaphagous Flies in Luoyang Based on 28S rRNA Gene Sequence / 法医学杂志

Objective To detect 715 bp sequence of 28S rRNA in sarcosaphagous flies, and to identify their common species for solving the problem of morphological identification, as well as providing technical support for postmortem interval (PMI) estimation. Methods Twenty-nine common sarcosaphagous flies were collected in Luoyang and classified by morphological characteristics. The DNA was extracted from the fly's legs by Chelex-100 method and then the fragments of 28S rRNA were amplified and sequenced. The results were compared with twenty-eight corresponding fly species of GenBank and EMBL databases. All the sequences were analyzed by MEGA7.0 software, and sequence alignment was performed by the searching in BLAST. The nucleotide composition was analysed, and the intraspecific and interspecific ge-netic distance and phylogenetic tree were established. Results Twenty-nine sarcosaphagous flies were classified into 6 species of 5 genera, 3 families by morphological characteristics. In the obtained 715 bp sequence of 28S rRNA , the comparison result of online BLAST showed that the similarity was 100％. Five species were well clustered by a phylogenetic tree. Between different groups, the interspecific and intraspecific differences ranged from 0.007 to 0.045 and 0 to 0.001, respectively. Conclusion The 28S rRNA target gene sequences shows a good identification capability, which can be a new genetic marker for the identification of sarcosaphagous flies.

Macrophages depletion impairs skeletal muscle regeneration by regulating inflammation and oxidative stress levels / 生理学报

The objective of this study was to explore the roles of macrophages in the regeneration of injured skeletal muscle and the mechanisms involved. Mice were randomly divided into the following groups: muscle contusion (S), muscle contusion control (S), macrophages depleted (T) and macrophages depleted control (T) groups. Muscle contusion model was created by high-energy blunt injury. Macrophages depletion model was constructed by injection of clodronate-liposomes. Their gastrocnemius muscles were harvested at the time points of 1, 3, 7 and 14 d post-injury. The changes in skeletal muscle morphology were assessed by hematoxylin-eosin (HE) staining and Masson's trichrome staining. The mRNA and protein levels of inflammatory cytokines, chemokines and oxidative stress factors were analyzed by real-time polymerase chain reaction (RCR) and Western blotting, respectively. HE staining results showed that a small amount of regenerating myofibers were observed in the S group (14 d post-injury), whereas a large number of regenerating muscle fibers were observed in the T group. Quantitative analyses showed that the sizes of regenerating myofibers were significantly smaller in the T group as compared with the S group at 14 d post-injury (P < 0.05). At the same time, Masson staining results showed that macrophage depletion significantly increased the area of fibrosis as compared with the S group at 14 d post-injury (P < 0.01). The expression levels of inflammatory cytokines, chemokines, and oxidative stress factors were increased significantly after muscle injury. Moreover, macrophage depletion increased the expressions of inflammatory cytokines, chemokines and oxidative stress factors as compared with the S group during the later stage of injury (7-14 d post-injury). These results suggest that macrophages depletion can aggravate fibrosis and impair muscle regeneration, and inflammatory cytokines, chemokines and oxidative stress factors may be involved in this process.

Relationship between Early Postmortem Interval and Electrical Conductivity of Rat Liver, Spleen and Kidney after Death in Rats / 法医学杂志

OBJECTIVES@#To determine the electrical conductivity （EC） of the liver, spleen and kidney of rats at different postmortem intervals （PMIs） within 24 hours for investigating the relationship between EC of different organs and early PMI.@*METHODS@#Totally 45 SD rats were sacrificed by cervical dislocation and kept at a constant temperature of 25 ℃. Tissues were taken from the liver, spleen, and kidney of rats at 0, 3, 6, 9, 12, 15, 18, 21 and 24 h. Impregnating solution with a mass concentration 0.1 g/mL was prepared using deionized water. The EC value of impregnating solution with different organs was separately determined. The regression equations of EC and PMI for different organs were established, respectively. The relationship between EC of different organs and early PMI was analysed in deceased rats.@*RESULTS@#The relationship between PMI and EC of the liver and spleen was well fitted with the linear equation. The liver showed the best fitting degree followed by the spleen, while the EC of the kidney showed no significant changes within 24 h. There was a good linear relationship between early PMI and the EC of the liver and spleen.@*CONCLUSIONS@#A good linear relationship between early PMI and the EC of the liver and spleen can be found in rats after death, which can be used for the early PMI estimation.

Virtual screening of anti-angiogenesis flavonoids from Sophora flavescens / 中国中药杂志

Angiogenesis is a dynamic, multi-step process. It is known that about 70 diseases are related to angiogenesis. Both the experimental and the literature reports showed that Sophora flavescens inhibit angiogenesis significantly, but the material basis and the mechanism of action have not been clear. In this study, molecular docking was used for screening of anti-angiogenesis flavonoids from the roots of S. flavescens. One handred and twenty-six flavonoids selected from S. flavescens were screened in the docking ligand database with six targets(VEGF-a,TEK,KDR,Flt1,FGFR1 and FGFR2) as the receptors. In addition, the small-molecule approved drugs of targets from DrugBank database were set as a reference with minimum score of each target's approved drugs as threshold. The LibDock module in Discovery Studio 2.5 (DS2.5) software was applied to screen the compounds. As a result, 37 compounds were screened out that their scores were higher than the minimum score of approved drugs as well as being in the top of 10%. At last the mechanism of flavonoids anti-angiogenesis was preliminarily revealed, which provided a new method for the development of angiogenesis inhibitor drugs.

Relationship between Electrical Conductivity and Decomposition Rate of Rat Postmortem Skeletal Muscle / 法医学杂志

OBJECTIVES@#To analyze the relationship among electrical conductivity （EC）, total volatile basic nitrogen （TVB-N）, which is an index of decomposition rate for meat production, and postmortem interval （PMI）. To explore the feasibility of EC as an index of cadaveric skeletal muscle decomposition rate and lay the foundation for PMI estimation.@*METHODS@#Healthy Sprague-Dawley rats were sacrificed by cervical vertebrae dislocation and kept at 28 ℃. Muscle of rear limbs was removed at different PMI, homogenized in deionized water and then skeletal extraction liquid of mass concentration 0.1 g/mL was prepared. EC and TVB-N of extraction liquid were separately determined. The correlation between EC (x₁) and TVB-N (x₂) was analyzed, and their regression function was established. The relationship between PMI (y) and these two parameters were studied, and their regression functions were separately established.@*RESULTS@#The change trends of EC and TVB-N of skeletal extraction liquid at different PMI were almost the same, and there was a linear positive correlation between them. The regression equation was x₂=0.14x₁-164.91(R²=0.982). EC and TVB-N of skeletal muscle changed significantly with PMI, and the regression functions were y=19.38x₁³-370.68x₁²+2 526.03 x₁-717.06(R²=0.994), and y=2.56x₂³-48.39x₂²+330.60x₂-255.04(R²=0.997), respectively.@*CONCLUSIONS@#EC and TVB-N of rat postmortem skeletal muscle show similar change trends, which can be used as an index for decomposition rate of cadaveric skeletal muscle and provide a method for further study of late PMI estimation.

Morphological and molecular identification of two strains of dermatophytes / 华中科技大学学报（医学）（英德文版）

In this study, we used traditional morphological and molecular identification methods to preliminarily identify two strains of dermatophytes. The two strains were observed under the microscope. And then the dermatophytes were cultured on Sabouraud's dextrose agar (SDA). The 18S rRNA regions of the two dermatophyte strains were amplified by polymerase chain reaction (PCR), and the PCR products were sequenced and compared with GenBank data. BLAST tools and DNAMAN software were used to analyze the sequences. To further determine highly homologous sequences, a phylogenetic tree was constructed using the Neighbor-Joining method. The two strains of dermatophytes were identified by traditional morphological identification as Epidermophyton floccosum and Microsporum ferrugineum. The 18S rRNA sequence analyses showed high similarities to Cladosporium cladosporioides isolate C115LM-UFPR and Ascomycete sp. LB68A1A2. Epidermophyton and Cladosporium belong to dermatophyte, while Microsporum ferrugineum and Ascomycete belong to microsporum. The two novel strains of dermatophytes were therefore identified as Cladosporium cladosporioides isolate C115LM-UFPR (JN650537, Cladosporium) and Ascomycete sp. LB68A1A2 (AY770409, Ascomycete sp).

Morphological and molecular identification of two strains of dermatophytes / 华中科技大学学报（医学）（英德文版）

In this study, we used traditional morphological and molecular identification methods to preliminarily identify two strains of dermatophytes. The two strains were observed under the microscope. And then the dermatophytes were cultured on Sabouraud's dextrose agar (SDA). The 18S rRNA regions of the two dermatophyte strains were amplified by polymerase chain reaction (PCR), and the PCR products were sequenced and compared with GenBank data. BLAST tools and DNAMAN software were used to analyze the sequences. To further determine highly homologous sequences, a phylogenetic tree was constructed using the Neighbor-Joining method. The two strains of dermatophytes were identified by traditional morphological identification as Epidermophyton floccosum and Microsporum ferrugineum. The 18S rRNA sequence analyses showed high similarities to Cladosporium cladosporioides isolate C115LM-UFPR and Ascomycete sp. LB68A1A2. Epidermophyton and Cladosporium belong to dermatophyte, while Microsporum ferrugineum and Ascomycete belong to microsporum. The two novel strains of dermatophytes were therefore identified as Cladosporium cladosporioides isolate C115LM-UFPR (JN650537, Cladosporium) and Ascomycete sp. LB68A1A2 (AY770409, Ascomycete sp).

Value of CK20 immunocytochemistry as an adjuvant to urine liquid-based cytology in detection of urothelial carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the value of cytokeratin 20 (CK20) immunocytochemical (ICC) detection in the urine liquid-based cytological specimens in diagnosis of urothelial carcinoma (UC).</p><p><b>METHODS</b>The study consisted of prospective and retrospective groups. In the prospective group, voided urine samples were collected from patients with a variety of urological conditions and healthy individuals. Urine cytological diagnosis and CK20 ICC were performed on the collected specimens. In the retrospective group, archived urine slides with cytological diagnoses of atypical urothelial cells (AUC), suspicious carcinoma (SuCA) and carcinoma (CA) were selected. Then they were re-stained immunocytochemically with monoclonal antibody against CK20 after decolorization. Histological diagnosis and clinical follow-up result were used as the gold standard for analysis.</p><p><b>RESULTS</b>There were 136 cases in the prospective group, including 89 cases of UC, 19 cases of other urogenital malignancies, 12 cases of benign lesions and 16 cases of normal control. The sensitivity of CK20 ICC in detection of UC was 75.3%, significantly higher than that of LBC (48.3%, P < 0.001). The positive rate of CK20 was 64.7% (22/34) in G1 UC, 73.3% (22/30) in G2 UC, and 91.3% (21/23) in G3 UC (P < 0.001). The specificity of CK20 ICC was 91.5%, the same as that of LBC. There were 163 cases in retrospective group, including 119 cases of UC, 17 cases of other urogenital malignancies and 27 cases of benign lesions. The cytological diagnoses of them were 68 cases of CA, 47 cases of SuCA and 48 cases of AUC. The positive rates of CK20 ICC in UC and non-UC (other urogenital malignancies and benign lesions) cases were 90.8% and 15.9%, respectively, with a statistically very significant difference (P < 0.001). The LBC of all the 119 cases of UC included 62 (52.1%) cases of CA, 35 (29.4%) cases of SuCA and 22 (18.5%) cases of AUC. The positive rates of CK20 in the LBC-diagnosed CA, SuCA and AUC were 96.8%, 97.1% and 63.6%, respectively. The LBC of all the 44 non-UC cases included 6 (13.6%) cases of CA, 12 (27.3%) cases of SuCA and 26 (59.1%) cases of AUC, and the positive rates of CK20 in the LBC-diagnosed CA, SuCA and AUC were 33.3%, 33.3% and 3.8%, respectively. The differences of UC and non-UC cases between the corresponding categories of LBC were significant (P < 0.0001, respectively).</p><p><b>CONCLUSION</b>CK20 immunocytochemistry as an auxiliary method to urine liquid-based cytology can increase the sensitivity in detection of urothelial carcinomas.</p>

Expression of EGFR, survivn and hnRNPA2/B1 in bronchial brushing cytology specimens and their significance in diagnosis of lung cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the expression of EGFR, survivn and hnRNPA2/B1 proteins in ThinPrep bronchial brushing cytology specimens and their significance in diagnosis of lung cancer.</p><p><b>METHODS</b>The protein expression of EGFR, survivn and hnRNPA2/B1 was detected by immunocytochemistry (ICC) in the specimens from 110 cases of non-small cell lung cancer (NSCLC), 32 cases of small cell lung cancer (SCLC) and 37 cases of non-neoplastic lung lesions. The relationship between EGFR, survivn and hnRNPA2/B1 protein expression and clinical characteristics of the patients was analyzed using SPSS 16.0 software.</p><p><b>RESULTS</b>(1) Positive expression was observed in 81.1% of NSCLC for EGFR, 66.2% for survivn, 90.9% for hnRNPA2/B1, significantly higher in NSCLC than in the control specimens (P = 0.000, P = 0.000, P = 0.010). The positive expression rate of hnRNPA2/B1 in SCLC was 92.3%, significantly higher than that in the control specimens (P = 0.021). (2) The expression of EGFR was associated with differentiation (P = 0.003), clinical stage (P = 0.023) and lymph node metastasis (P = 0.038), but was not associated with gender, age and histological types. The survivn expression was not related with the above mentioned clinicopathological features (P>0.05). Expression of hnRNPA2/B1 was associated with clinical stage (P = 0.017), but not associated with gender, age, histological type, differentiation and lymph node metastasis. (3) There was a significant difference between the co-expression of EGFR and survivn in NSCLC (98.0%) and benign conditions (2.0%, P = 0.000), also a significant difference between the negative expression of both EGFR and survivn in NSCLC (38.2%) and nonneoplastic lesions (61.8%, P = 0.000).</p><p><b>CONCLUSIONS</b>Analysis of EGFR, survivn and hnRNPA2/B1 expression may be an useful adjunct method to stratify controversial cases. The positive expression of EGFR might be associated with invasion, progression and poor prognosis of NSCLC.</p>

Comparison of touch imprint cytology of core needle biopsy and section histopathology in breast cancer diagnosis / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the sensitivity, specificity of touch imprint cytology (TIC), and to compare its conformity rate with histopathology, to observe the consistence of immunocytochemistry (ICC) with immunohistochemistry (IHC), and to assess the diagnostic value of TIC prior to neoadjuvant chemotherapy for breast cancer.</p><p><b>METHODS</b>289 cases of TIC and 287 cases with core needle biopsy (CNB) histopathology accumulated from October 2005 to October 2008 in our hospital were included in this study. One hundred ninety cases TIC results were compared with that of final histopathology. 64 cases were tested for ER, PR, HER-2 by immunocytochemistry.</p><p><b>RESULTS</b>Twenty-four benign cases and 263 malignant cases were diagnosed. 4 specimens were unsatisfactory. False negative rate and unsatisfactory rate were 1.4%, both, and false positive rate was 0.35%. The accuracy rate of TIC and CNB was 95.8% and 95.3%, respectively (P = 0.804). The sensitivity of TIC and CNB was 96.2% and 95.0% (P = 0.601), specificity 87.5% and 100% (P = 0.471) were found, when compared with the results of routine histopathology. 52 cases had a control with IHC of CNB in 64 ICC, and 43 cases had a final histopathology IHC. The ICC conformity rate of ER, PR, HER-2 with IHC of CNB was 86.5%, 75.0%, 78.8%, and that with IHC of final histopathology was 88.4%, 74.4%, 75.6%, respectively. The conformity rate of IHC between CNB and final histopathology was 83.7%, 74.4%, 76.5%, respectively. There was no significant statistical difference between them.</p><p><b>CONCLUSION</b>Compared with routine CNB histopathology, TIC has a high accuracy and sensitivity, and can provide a rapid and reliable cytological diagnosis to complement CNB for breast lesions. The conformity rates are high in ER, PR, HER-2 expression between ICC and IHC. ICC of TIC can be used to determine the estrogen and progesterone receptor levels in breast cancer before neoadjuvant chemotherapy.</p>

The value of thin-layer cytology in intraoperative fine-needle aspiration biopsy of pancreatic cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The purpose of this study was to explore the value of thin-layer cytology (TLC) in intraoperative fine needle aspiration cytology diagnosis of pancreatic cancer.</p><p><b>METHODS</b>Results of cytological examination with thin-layer smears were compared with that with conventional smears in intraoperative fine-needle aspiration (FNA) biopsy.</p><p><b>RESULTS</b>Totally 271 fine needle aspiration biopsies were performed, among them, 70 were examined with thin-layer smears, showing unsatisfactory smear in 5 cases (7.1%); 201 were examined with conventional smears (CS), showing unsatisfactory smear in 9 cases (4.5%). No significant difference in the unsatisfactory smears was observed between those two groups. The positive rate of diagnosis with CS smears was 60.0% (42/70) and that of TLC was 81.6% (164/201), with a significant difference (P < 0.01). The sensitivity of CS and TLC was 68.9% and 87.7%, respectively (P < 0.01). The sensitivity of both FNA and frozen section diagnosis in 20 cases was 90.0%, respectively, but that of FNA combined with frozen section diagnosis was 95.0%. 9 cancer cases diagnosed by pathology were initially negative by cytology, but adenocarcinoma cells were found in 7 cases of them by the second time cytology examination.</p><p><b>CONCLUSION</b>The positive rate is high in TLC smears, and unsatisfactory rate is low. TLC smears are one of the best methods in intraoperative confirmation of pancreatic cancer. The use of FNA smears combined with frozen section biopsy can further improve the sensitivity of diagnosis.</p>

Application of protein markers in combination with ThinPrep bronchial brush cytology in classification of lung cancer subtypes / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the value of application of cellular protein markers stained by immunocytochemistry in combination with ThinPrep bronchial brush cytology in classification of lung cancer subtypes.</p><p><b>METHODS</b>Remaining bronchial brush cytology samples from 206 lung cancer patients with positive cytological diagnosis and 45 fine needle aspiration samples of resected lung carcinomas were collected. The expressions of CK10/13, CK7, CK18, CD56 and SYN in those samples were detected by immunocytochemistry (ICC) using corresponding antibodies.</p><p><b>RESULTS</b>The sensitivity and specificity of CK10/13 were 94.7% and 72.0%, respectively, in diagnosis of squamous cell carcinoma. The sensitivity and specificity of CK7 were 98.6% and 61.5%, and those of CK18 were 98.6% and 37.5%, respectively, in diagnosis of adenocarcinoma. The sensitivity and specificity of CD56 were 86.3% and 82.9%, and those of SYN were 81.6% and 93.5%, respectively, in diagnosis of small cell lung cancer. No significant difference was found in the expressions of CK10/13, CK7 and CK18 protein markers among differently differentiated lung squamous cell carcinomas and adenocarcinomas (P > 0.05). The classification rate of cytology in combination with ICC in differential diagnosis for 44 cases of unclassified lung cancer reached 90.0% for squamous cell carcinoma, 96.3% for adenocarcinoma, and 100.0% for small cell lung carcinoma.</p><p><b>CONCLUSION</b>Application of cellular protein markers in combination with ThinPrep bronchial brush cytology is helpful to improve the differential diagnosis of lung cancer subtypes, and may become a supplementary diagnostic method in subclassification of lung cancer.</p>

Study on stability of curcumine, demethoxycurcumin and bisdemethoxycurcumin / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the stability of curcumin, demethoxycurcumin and bisdemethoxycurcumin in different buffer solution.</p><p><b>METHOD</b>To determine concentration of curcumin by HPLC when added curcumin, demethoxycurcumin and bisdemethoxycurcumin into the buffer solution the equation of degradation was established.</p><p><b>RESULT</b>The sequence of stability are bisdemethoxycurcumin > or = demethoxycurcumin > or =curcumin at the same condition.</p><p><b>CONCLUSION</b>The demethoxycurcumin can stabilize curcumin more strong than the others. The demethoxycurcumin is a nature stabilizing agent for curcumin.</p>