[Role of intercellular adhesion molecule-1 in adhesion and migration of mesenchymal stem cells in ankylosing spondylitis and its mechanisms].

Objective: To investigate the role and underlying mechanisms of intercellular adhesion molecule-1 (ICAM-1) in the adhesion and migration of mesenchymal stem cells (MSCs) in patients with ankylosing spondylitis (AS). Methods: Bone marrow and ligament tissues were collected during surgery from patients with AS and thoracolumbar fractures (as controls, HC) treated from October 2021 to October 2022 at Nanjing University Medical School Affiliated Nanjing Drum Tower Hospital. MSCs were isolated and cultured from the bone marrow using the Ficoll separation method. Cell morphology was observed under high-resolution microscopy, and differences in the cytoskeletal features between AS-and HC-MSCs were analyzed through immunofluorescence staining. The expression of ICAM-1 was quantified in both groups using real-time quantitative polymerase chain reaction (RT-qPCR) and flow cytometry. Transwell migration assays and wound healing experiments were conducted to evaluate the differences in migration rates between the two groups of MSCs. Results: The interspinous ligament and bone marrow was acquired in AS (2 males and 1 female; 33, 37, 32 years old, respectively) and no-AS patients (2 males and 1 female; 35, 32, 38 years old, respectively). AS-MSCs exhibited broader cell morphology compared to HC-MSCs under bright field and fluorescence microscopy. Immunofluorescence staining of the interspinous ligament showed higher expression of ICAM-1 (68.38±3.42 vs 48.31±2.43) and CD105 (37.97±2.16 vs 23.36±2.06) in AS patients (both P<0.001). Western blot and RT-qPCR analysis revealed significantly stronger protein expression and transcription levels of ICAM-1 in AS-MSCs when compared to those in HC-MSCs (both P<0.001). Flow cytometry confirmed greater mean fluorescence intensity of ICAM-1 in AS-MSCs than in that in HC-MSCs (924.30±54.99 vs 636.47±40.03, P=0.002). Regarding cell adhesion efficiency, it showed no significant difference between AS-MSCs and HC-MSCs in the early stage of adhesion (0.5 h: 1 496±213 vs 1 205±163, P=0.133), but they were all significantly higher in AS-MSCs in the later stage (1 h: 2 894±172 vs 1 908±155, P=0.002; 2 h: 4 540±286 vs 3 334±188, P=0.004; 3 h: 5 212±281 vs 4 208±303, P=0.014). Finally, cell migration experiments demonstrated a stronger migration capability of AS-MSCs compared to HC-MSCs (5 449±172 vs 4 016±155, P<0.001), and the inhibition efficiency of A-205804 on the migration rate of AS-MSCs was stronger than that on HC-MSCs (2 145±239 vs 3 539±316, P=0.004). Conclusions: The aberrant expression of ICAM-1 markedly influences the adhesion and migration dynamics of MSCs. Elevated ICAM-1 levels in MSCs derives from patients with AS significantly enhance their migratory capabilities.

Serum matrix metalloproteinases-3 levels in patients with ankylosing spondylitis.

Cumulated evidence indicates that matrix metalloproteinase-3 (MMP-3) is significantly involved in cancer progression. Recent studies yielded conflicting results regarding the association between serum MMP-3 and ankylosing spondylitis (AS). To clarify this correlation, we performed a meta-analysis. Potential relevant studies were identified by searching the following databases: PubMed, Embase, CINAHL, Science Citation Index database, the Cochrane Library, Current Contents Index, Chinese Biomedical, the Chinese Journal Full-Text, and the Weipu Journal. Data from eligible studies were extracted and included into the meta-analysis using a random-effect model. Standardized mean differences (SMDs) and 95% confidence intervals (CIs) were used to assess the association between serum MMP-3 levels and AS. Thirteen case-control studies, including 707 AS cases and 442 healthy controls, were selected for the meta-analysis. The results indicate a significantly higher serum MMP-3 level in patients with AS than that in the controls (cases vs controls: SMD = 1.31, 95%CI = 0.84-1.78, P < 0.001). Ethnicity-subgroup analysis indicated a higher MMP-3 level in Asian and Caucasian patients with AS (all P < 0.05). This meta-analysis indicates that increased serum MMP-3 level correlates with the development of AS, suggesting that MMP-3 may present a clinical value in reflecting the progression of AS. Further larger sample size studies are warranted.

Weekly paclitaxel plus carboplatin is an effective nonanthracycline-containing regimen as neoadjuvant chemotherapy for breast cancer.

BACKGROUND: To evaluate the activity and safety of nonanthracycline-containing weekly PCb [paclitaxel (Taxol) plus carboplatin] regimen in neoadjuvant treatment of breast cancer. PATIENTS AND METHODS: Eligible patients were assigned to receive four cycles of PCb with dose of paclitaxel 80 mg/m(2) and carboplatin at an area under the curve of 2 mg x min/ml, given day 1, day 8 and day 15 of every 4 weeks. Pathological complete remission (pCR) was defined as no invasive cancer in breast and axillary samples. RESULTS: Overall, 107 consecutive patients received weekly PCb treatment from December 2007 to December 2008, and one was diagnosed with bilateral breast cancer. A total of 85.2% of patients were initially diagnosed with stage III diseases. Clinical response rate was 86.1% with complete remission rate 32.4%. Twenty-one patients achieved pCR after neoadjuvant treatment, with pCR rate 19.4%. The incidence of grade 3-4 neutropenia was 40.2% and only one patient was reported with febrile neutropenia. Severe anemia and thrombocytopenia occurred in 4.7% and 0.9%, respectively, of patients. Peripheral neuropathy was frequent but never severe. Patients with estrogen receptor-negative, progesterone receptor-negative, triple-negative or human epidermal growth factor receptor 2 (Her2)-positive subtype disease had higher pCR. CONCLUSIONS: Weekly PCb regimen was very active and tolerable as neoadjuvant treatment of breast cancer. This weekly PCb regimen should consider as a reasonable nonanthracycline-containing option in the neoadjuvant treatment of breast cancer.

Effects of lncRNA DANCR on proliferation and differentiation of osteoblasts by regulating the Wnt/ß-catenin pathway.

OBJECTIVE: To measure the expression level of long non-coding ribonucleic acids (lncRNAs) differentiation antagonizing non-protein coding RNA (DANCR) in serum of patients with fracture and investigate its influences on the proliferation and differentiation of osteoblasts. PATIENTS AND METHODS: Serum samples were collected from 44 fracture patients treated in our hospital and 24 healthy people receiving physical examination in our hospital. Then, reverse transcription-polymerase chain reaction (RT-PCR) technique was used to detect the expression of lncRNA DANCR in the serum of patients with fracture and healthy subjects. MC3T3-E1 mouse osteoblast cell line with stably-knocked out DANCR was further established using small interfering RNAs (siRNAs), and the effect of DANCR knockout on the proliferation of osteoblasts was determined using cell counting kit-8 (CCK-8). At the same time, 5-Ethynyl-2'-deoxyuridine (EdU) staining assay was performed to detect the percentage of EdU-positive cells in osteoblasts in control group and DANCR knockout group. In addition, the mRNA levels of differentiation-related genes including Runt-related transcription factor 2 (Runx2), Collagen1α1, osteocalcin (OC) and osterix (OSX) were detected via RT-PCR, and the protein level of Runx2 was measured through Western blotting. Moreover, osteoblasts were cultured with osteogenic medium for 14 d, and then alizarin red staining and alkaline phosphatase (ALP) staining assays were carried out to examine the differentiation of these osteoblasts. Lastly, Western blotting technique was employed to analyze the expression of the Wnt/ß-catenin signaling pathway. RESULTS: The expression of lncRNA DANCR was significantly increased in the serum of fracture patients (p<0.05). The results of in-vitro cell experiments showed that the intervention of DANCR with siRNA was able to clearly promote the proliferation and differentiation of MC3T3-E1 osteoblast cell line. According to the results of Western blotting, DANCR promoted the apoptosis and proliferation, which was mediated by the activated Wnt/ß-catenin signaling pathway in osteoblasts. CONCLUSIONS: LncRNA DANCR inhibition can facilitate the proliferation and differentiation of osteoblasts by activating the Wnt/ß-catenin signaling pathway in osteoblasts. Therefore, DANCR is expected to be a new target promoting fracture healing.

CoV2O4: a novel anode material for lithium-ion batteries with excellent electrochemical performance.

This study reports the electrochemical applications of CoV2O4 as a novel anode for lithium-ion batteries. Ex situ analyses were performed to understand the conversion that transpires during the charge-discharge cycle. Also, the effects of different binders were analyzed. With the synergistic effect of Na-carboxymethyl cellulose and styrene butadiene rubber (CMC and SBR), a reversible capacity of 727.5 mA h g-1 was obtained after 100 cycles which indicates potential applications of CoV2O4 in energy storage devices.

Over expression of aromatase protein is highly related to MMPs levels in human breast carcinomas.

Estrogens can play a critical role in the development of breast cancer. Aromatase which catalyzes the formation of aromatic C18 estrogens from C19 androgens is regarded to be responsible for the cancer local production of estrogen. Studies not only from aromatase transfected breast cancer cells, but also transgenic mouse, which overexpressed aromatase, demonstrated that in situ produced estrogen plays more important roles than circulating estrogens in breast tumor promotion and progression. Matrix-metalloproteinases (MMPs) have been implicated in the proeolytic process, which play important roles in the aggressiveness of cancer cells including invasion of adjacent tissue and metastasis to distant sites. Expression of MMP2 and 9 may be stimulated by estrogens in hormonal dependent breast cancers, since tumor aromatase can stimulate breast cancer growth and progression in both an autocrine and a paracrine manner. Theoretically aromatase overexpression, that causes relatively high estrogen concentration in situ, may be positively related to MMP2 and 9 expression, indicate worse prognosis in breast cancers, and maybe insensitive to tamoxifen therapy. In the present study, we studied the expression of aromatase activity and MMPs in human breast carcinoma both in vitro and in vivo. In human breast carcinoma cell lines including MCF-7, MDA-MB-231 and MDA-MB-435, the expression of aromatase levels both in mRNA and protein activity was related to MMP2 and MMP9. In humam breast cancer samples, we demonstrated that aromatase expressions were strongly associated with MMP2 and MMP9 levels. It was interesting to observe that the positive relationship was only present in the ER and/or PR positive patients. This may indicate that both MMP2 and MMP9 were up regulated by estrogen produced by aromatase through ER. So in endocrine therapy, either blocking the ER by tamoxifen or inhibiting the aromatase by aromatase inhibitors for example letrozole, may both inhibit tumor growth and lower the metastatic potential especially in ER positive breast cancer patients by means of down-regulation of MMP2/9.

Fiberoptic ductoscopy for breast cancer patients with nipple discharge.

BACKGROUND: Breast cancer and precancer are thought to originate in the lining of the milk duct, but until recently, we have not had direct access to this area other than in tissue removed blindly by core biopsy or fine-needle aspiration. Fiberoptic ductoscopy (FDS) is an emerging technique that allows direct visual access of the ductal system of the breast through nipple orifice cannulation and exploration. To date, this technique has been used only in pilot studies. Previously, we have demonstrated that fiberoptic ductoscopy in patients with and without nipple discharge is a safe and effective means of visualizing the intraductal lesion. When combined with cytology, it is a screening technique that has high predictive value. METHODS: We applied ductoscopy to 415 women with nipple discharge with the specific intent of detecting those patients with nipple discharge who had intraductal carcinoma (DCIS) as the basis of their discharge. RESULTS: In this cohort of patients, ductoscopy was successful in visualizing an intraductal lesion in 166 patients (40%). In these cases, ductal lavage following ductoscopy increased the yield of cytologically interpretable ductal epithelial cells 100-fold compared to discharge fluid alone. In the majority of these patients, FDS examination detected lesions that had the appearance of typical papillomas. However, in 10 patients, the intraductal lesion exhibited one of several atypical features, including bleeding, circumferential obstruction, and gross fungating projections. In eight of these patients, the subsequent histopathology turned out to be DCIS. In two of these eight patients, endoscopic biopsy revealed cytologically malignant cells; in two others, ductal lavage (washings) revealed cytologically malignant cells. In three additional patients, although FDS examination uncovered a typical papilloma that was not biopsied, ductal lavage (washings) revealed cytologically malignant cells. On surgical pathology review of the extirpated lesions, all 11 patients were subsequently shown to have DCIS. Of these 11 cases of DCIS that were initially detected with a combination of FDS and ductal lavage cytology, six were completely negative on mammogram and physical exam. CONCLUSION: Although nipple discharge is an unusual presentation for DCIS, in patients with nipple discharge, FDS with ductal lavage cytology is a useful technique for diagnosing DCIS prior to definitive surgery.

Pregnancy-associated breast cancer: a case control and long-term follow-up study in China.

Pregnancy associated (PA) breast cancer is defined when diagnosed during pregnancy or within one year afterwards. To analyze the prognostic factors related to this disease and assess the impact of pregnancy on breast cancer patients, 88 PA breast cancer patients initially treated in the Shanghai Cancer Hospital from 1957 to 1990 were reviewed. A non-PA group including 176 patients individually well matched to the PA group was also analyzed. Univariate analysis suggested prognostic value for clinical tumor size, TNM stage, and breast feeding time among the classic prognostic factors, pregnancy associated factors and treatment modalities were evaluated. Multivariate analysis demonstrated clinical tumor size, TNM stage and axillary lymph node metastasis as independent prognostic factors. Compared with the non-PA group, the PA group was significantly correlated with delay at diagnosis, large size of the tumor, late TNM stage, extension to the skin or chest wall and administration with oophorectomy. The overall survival rates of 5-year, 10-year and 20-year were 40.39%, 36.29% and 30.70%, respectively, which were worse than those in the non-PA group, but did not reach a significant difference (p=0.0536). We are conducting further basic research to analyze the biologic characteristics of PA breast cancer.

Reduction of myocardial ischemia-reperfusion injury by isovolumic hemodilution.

In order to study the effects of isovolumic hemodilution and its combination with Danshen solution on acute ischemic reperfused canine myocardium, 24 adult hybrid dogs were used and divided into four groups. Group I was the control group, groups II-IV were treated with Dextron 40, Danshen solution and a combination of the two, respectively. The results showed that either Dextron 40 or Danshen solution alone had a significant increase of +/- dp/dt-max when compared with group I (p < 0.05). Although no significant difference existed between group II and III, the former showed more rapid action. The combination of the two therapies improved +/- dp/dt-max and LVSP, and significantly reduced the necrotic sizes and the MDA contents in the ischemic myocardia compared with not only group I, but also group II or group III (all p < 0.05). The results suggest that isovolumic hemodilution or Danshen may protect the ischemic reperfused myocardium and the former may come into action more rapidly, and that the combination of the two may show a better synergism than each one of the two by itself.

[Role of pp60c-src in mitogen-activated protein kinase activation of vascular smooth muscle cells].

The effect of pp60c-src on mitogen-activated protein kinase (MAPK) activation induced by angiotensin II (Ang II) in vascular smooth muscle cells (VSMCs) was observed to explore the signal transduction process of VSMCs proliferation mediated by Ang II. Following transfection of the synthetic antisense c-src oligodeoxynucleotides (ODNs) wrapped in lipofectin into cultured rat VSMCs, pp60c-src content markedly decreased, pp60c-src kinase activity was only 14.4% of control, while MAPK activity was not significantly changed (P>0.05). When these transfected VSMCs were further stimulated by Ang II, the increase in MAPK activity was only 1.6% of control. These data suggest that pp60c-src is needed for MAPK activation in VSMCs stimulated by Ang II, and pp60c-src may be an important signaling molecule in abnormal proliferation of VSMCs.

[Timing of radical operation on Hirschsprung's disease in newborns].

From June 1987 to June 1993, 42 neonates with Hirschsprung's disease (NHD) were treated with modified Duhamel's procedure. The follow-up from 2 months to 5 years (mean 30 mons) showed satisfactory results in all survivors. Comparing with 21 infants with Hirschsprung's disease treated (IHD) with modified Swenson's method showed that: (1) The follow-up results of NHD were better than IHD; (2) The extent and degree of hypertrophy and dilatation of proximal and transitional segment were less serious in NHD and the length of resection was shorter than that of IHD (NHD: 14.5-24.1 cm, IHD: 33-40.5 cm); (3) The degree of pathological changes such as mucosal inflammation, ulcer formation, and degeneration of ganglion cells was related to the age, duration and the length of agangliosis. Hirschsprung's disease has a chronic progressive course. Early operation can decrease the secondary structural and functional damage to the intestine.

Effects of tetrandrine on rabbit platelet aggregation and platelet activating factor generation.

AIM: To study the effects of tetrandrine (Tet) on platelet aggregation and platelet activating factor (PAF) generation in rabbit platelet-rich plasma (PRP). METHODS: The aggregation rate of platelets induced by calcimycin (Cal) and PAF and the inhibition rate of Tet on platelet aggregation were measured. The amount of PAF in PRP stimulated with Cal and treated with Tet was also measured. RESULTS: Cal 1-8 mumol.L(-1) and PAF 9.5-190.5 pmol.L(-1) induced platelet aggregation. At the final concentrations of 4-64 mumol.L(-1), Tet inhibited the aggregation induced by Cal 4 mumol.L(-1) and PAF 142.9 pmol.L-1. The IC50 (95% confidence limits) were 8.6 (6.0-12.2) mumol.L-1 for Cal and 14.0 (6.4-30.4) mumol.L(-1) for PAF. In the PRP aggregation by Cal, there was a marked increase in PAF content. Tet dependented the release of PAF from platelets by Cal in a concentration-dependent manner, with IC50 of 21 (8-54) mumol.L(-1). CONCLUSION: The inhibition effect of Tet on platelet aggregation might be concerned with the reduction of endogenous PAF generation.

[Immunologic histochemical observation of anti-crypt keratin of cryptic epithelium of tonsilla in children with chronic tonsillitis].

OBJECTIVE: To investigate anti-crypt keratin (CK) immunologic histochemical changes in children with chronic tonsillitis. METHOD: Removed tonsilla were fixed by 10% formaldehyde. Immunologic histochemical method was used to determine the changes of anti-broad spectrum (KD 68, 56, 56, 50) CKSP. RESULT: In 230 cases, obvious keratosis was 90.9%, no keratosis was 9.1%, 3 cases were found with fungus filaments and bacteria in the bottom of crypts. Anti-broad spectrum and hypermolecule CK of tonsil cryptic epithelium were positive reaction, anti-broad spectrum CK of cryptic keratosis in all cases was positive reaction. CONCLUSION: During the period of episode, cryptic epitheliums of tonsilla was destroyed repeatly, therefore, immunoglobulin production was reduce. Because the immune function of tonsilla was reduced, bacteria and virus might be invade into organism. This reduplicative malignant circles must be interrupted or blocked only by tonsillectomy.

Fiberoptic ductoscopy for patients with nipple discharge.

BACKGROUND: Breast carcinoma and precancer are thought to start in the lining of the milk duct or lobule, yet until recently, we have not had direct access to this area other than by blindly removing tissue by core biopsy or fine-needle aspiration. Fiberoptic ductoscopy (FDS) is an emerging technique allowing direct visual access to the ductal system of the breast through nipple orifice exploration. METHODS: We applied ductoscopy to 259 women who had nipple discharge, and we analyzed the visual findings, the cytological washings, and the subsequent histopathology. RESULTS: In 92 (36%) of these women, fiberoptic ductoscopy was successful in detecting an intraductal papillary lesion. Of these observed lesions, 68 (74%) were single papilloma, 21 (23%) were multiple discrete papillomas, and 3 (3%) were diffuse intraductal thickening which corresponded to diffuse papillomatosis on histopathological analysis. The overall positive predictive value of FDS screening was 83%. Of the lesions observed, 29.8% were located in the main (segmental) duct, 43.9% lesions in the first branch, 17.5% lesions in the second branch, 7.9% in the third branch, and 0.9% in the fourth branch. These lesions had an overall average distance of 2.7 cm from the nipple orifice. Ductal washings performed at the time of ductoscopy were effective at obtaining representative exfoliated ductal cells which could be evaluated for the presence of clumps (> 50 cells), clumps with atypia or single ductal cells. The presence of clumps with positive FDS increased the positive predictive value to 86%. CONCLUSIONS: Fiberoptic ductoscopy currently offers a safe alternative to ductography in guiding subsequent breast surgery in the treatment of nipple discharge.

p21/waf1/cip1 and mdm-2 expression in breast carcinoma patients as related to prognosis.

p21/waf1/cip1 and mdm-2 are downstream effectors of p53. p21 plays a major role in negatively regulating cell-cycle progression, while mdm-2 inhibits p53 effects, and its role has been implicated in oncogenesis. In this study, we investigated the expression profiles of p21, mdm-2 and p53 in human breast-carcinoma tissues. The aim was to determine whether a correlation exists between the expression profiles of these markers and tumor differentiation, ER status and prognosis. We studied tumor specimens obtained from 106 patients and found a highly significant association among low histology grade, p53 over-expression, high mdm-2 expression and lack of p21 expression. Our studies also demonstrate that, in human breast cancer, low levels of p21 and higher mdm-2 levels directly correlate with the onset of lymph-node metastases and shortened patient survival. Furthermore, the expression profiles of p21, mdm-2 and p53 were independently correlated with patient survival.

Prognostic role of p27Kip1 and apoptosis in human breast cancer.

Human breast carcinoma is biologically heterogeneous, and its clinical course may vary from an indolent slowly progressive one to a course associated with rapid progression and metastatic spread. It is important to establish prognostic factors which will define subgroups of patients with low vs high risk of recurrence so as to better define the need for additional therapy. Additional characterization of the molecular make-up of breast cancer phenotypes should provide important insights into the biology of breast cancer. In the present study, we investigated apoptosis, expression of p27Kip1 and p53 retrospectively in 181 human breast cancer specimens. In addition, their relevance to the biological behaviour of breast cancer was examined. Our studies found a significant association among high histological grade, high p53, low apoptosis and low p27. Our results also demonstrated that, in human breast cancer, low levels of p27 and apoptotic index (AI) strongly correlated with the presence of lymph node metastasis and decreased patient survival. In node-negative patients, however, p27 also had prognostic value for relapse-free and overall survival in multivariate analysis. Furthermore p27 and AI had predictive value for the benefits of chemotherapy. These latter observations should prompt prospective randomized studies designed to investigate the predictive role of p27 and AI in determining who should receive chemotherapy in node-negative patients.

Identification of a unique binding protein specific for a novel retinoid inducing cellular apoptosis.

The retinoid 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalenecarboxylic acid (AHPN, CD437) induces apoptosis in a variety of cell types, many of which are cancer cells that resist the antiproliferative and/or differentiating effects of retinoids. While the retinoids exert their effects by binding to the retinoic acid nuclear receptors (RARs) or retinoid X receptors (RXRs), AHPN (CD437) binds to another protein with different ligand specificity. In nuclear extracts from HL-60R cells the binding of AHPN (CD437) was only minimally competed by either retinoic acid (tRA)or 9-cis-retinoic acid (9-cis-RA), the natural ligands for the RARs and RXRs, respectively. Moreover, AHPN (CD437) was unable to compete with either tRA or 9-cis-RA for binding to endogenous retinoid receptors in nuclear extracts from the MDA-MB-468 breast carcinoma cell line. Size exclusion chromatography revealed AHPN binding to a 95 kDa protein(s) which is neither an RAR or RXR. Our results suggest that apoptosis induction by AHPN (CD437) may occur through interaction with another protein and is independent of the RAR/RXR-signaling pathways.