[Landscape and metastases of the lymph nodes in prostatic anterior fat pad at radical prostatectomy].

Objectives: To examine the landscape and metastases of the lymph nodes in prostatic anterior fat pad (PAFP) at radical prostatectomy (RP), and to describe the clinical characteristic of the patients with lymph node metastases in PAFP. Methods: The clinical and pathological data of 287 prostate cancer patients underwent RP from December 2019 to August 2021 in Department of Urology, Sun Yat-sen University Cancer Center were collected and analyzed retrospectively. All patients were male, aging (66±7) years (range: 42 to 83 years). The preoperative prostate-specific antigen (PSA) (M(IQR)) were 16.00(29.64) µg/L (range: 0.01 to 99.90 µg/L). There were 244 patients with localized or locally advanced prostate cancer and 43 patients with metastatic prostate cancer. All PAFP were dissected at RP routinely and were sent for pathologic analysis respectively. The PAFP was dissected from the prostate apex caudally toward the bladder neck and dissection extended to the joint of the prostate and the endopelvic fascia bilaterally. All the specimen of PAFP were examined and reported by subspecialty pathologists of genitourinary tumors. Statistical analysis was performed by Student t test, Wilcoxon rank-sum test, χ2 test or Fisher exact test. Results: There were 8.0% (23/287) patients with lymph nodes in PAFP, 3.8% (11/287) patients with PAFP lymph node metastases. Pathologically upstaged occurred in 1 patient due to the PAFP lymph node as the solitary metastatic lesion. Patients with lymph node metastases in PAFP presented higher preoperative PSA (M(IQR): 48.2(73.0) µg/L vs. 15.4(26.5) µg/L, Z=3.158, P=0.002), clinical T stage and N stage (Z=2.977, P=0.003; Z=2.780, P=0.005) and preoperative Gleason score (Z=2.205, P=0.027). Conclusions: Routine dissection of PAFP at RP and separately pathological analysis may allow more lymph nodes and lymph node metastases detection. More accurate pathological N stage may be acquired and consequently may improve the survival of patients by offering more appropriate adjuvant or salvage therapy.

[Correlation between the expression profile of cytokeratin 19/glypican 3 and recurrence of hepatocellular carcinoma after interventional therapy].

Objective: To observe the correlation between the expressions profile of cytokeratin 19/glypican 3 (CK19/GPC3) and recurrence of hepatocellular carcinoma after interventional therapy. Methods: Clinical and pathological information of 251 eligible cases with hepatocellular carcinoma who underwent interventional therapy in You'an Hospital from November 2007 to May 2016 were retrospectively collected. Univariate and multivariate Cox regression analysis was used to analyze the relevant risk factors that may affect their prognosis. Kaplan-Meier survival analysis was used to draw the survival curve. Log-rank test was used to compare the difference in survival rates between the groups. Results: Kaplan-Meier univariate analysis showed that histological grade, CK19/GPC3 expression profile, alpha-fetoprotein level and Hep Parl were closely related to tumor recurrence. Multivariate Cox regression analysis showed CK19/GPC3 expression profile (HR = 1.634, 95%CI: 1.041 ~ 2.564, P = 0.033), histological grade (HR = 1.445, 95%CI: 1.037 ~ 2.014, P = 0.030), alpha-fetoprotein level (HR = 1.410, 95%CI: 1.042 ~ 1.908, P = 0.026), Hep Parl (HR = 0.570, 95%CI: 0.349 ~ 0.930, P = 0.025) were the four independent factors for prediction of recurrence after interventional therapy. Conclusion: Hepatocellular carcinoma patients with CK19(+)/GPC3(+) and CK19(-)/GPC3(+) phenotypes who meet the Milan criteria have a higher risk of recurrence after interventional therapy than CK19(-)/GPC3(-) phenotypes.

In vivo study of microarc oxidation coated Mg alloy as a substitute for bone defect repairing: Degradation behavior, mechanical properties, and bone response.

Large segmental bone defect healing remains a great challenge in clinic. Limited by the source of autograft, bone graft substitute tends to be the research focus. In the present study, we propose a strategy by using microarc oxidation (MAO) coated magnesium scaffold as a large segmental bone graft substitute, utilizing its combination of strength, degradability, and controllable corrosion rate. Bare substrate, 10 µm and 20 µm thick MAO coated Mg scaffolds were implanted into ulna bone of New Zealand white rabbits, employing a 15 mm wide bone defect model. The biocompatibility and in vivo degradation of the implants, the bone defect healing response, and mechanical properties of the injured bone were investigated. The surface cytocompatibility evaluation results show that the MAO coated Mg are more suitable for cell proliferation. Micro-CT results show that abundant new bone formed and initially bridged the 15 mm gap at 8 weeks. Histological results indicate the newly formed bone was full of maturation at 12 weeks. Three point bending tests reveal that the injured bone possessed sufficient mechanical strength after 12 weeks. A 3-step in vivo degradation mechanism was proposed for the implants. In summary, we observed an actual trial of 15 mm wide bone defect healing where the newly formed bone bridged the bone gap at 8 weeks successfully. These data suggest a great potential of MAO coated magnesium to be a bone graft substitute.

MiR-30c exerts tumor suppressive functions in colorectal carcinoma by directly targeting BCL9.

OBJECTIVE: Colorectal carcinoma (CRC) remains a leading health threat worldwide due to its high mortality. MicroRNA (miR-30c) is an important tumor suppressor in various cancers. B cell lymphoma 9 (BCL9) is one of the candidate genes for cancers. The synergistic effects of miR-30c and BCL9 in CRC progression remain to be carefully elucidated. PATIENTS AND METHODS: Fifty pairs of CRC samples and matched adjacent non-tumor tissues were collected from Yantai Yuhuangding Hospital between 2015 and 2017. MiR-30c and BCL9 expression levels were measured by quantitative Real-time polymerase chain reaction (qRT-PCR) in CRC tissues and cell lines. The 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to determine the influence of miR-30c on the proliferation ability of CRC cells. Target Scan was used to predict the potential target of miR-30c. Then, luciferase assay was performed to confirm the prediction. In addition, we also investigated the biological influence of BCL9 on miR-30c-mediated functions in CRC. RESULTS: We found that miR-30c was significantly decreased in CRC tissues and cell lines while the BCL9 expression level was prominently increased in CRC tissues and cells. Additionally, the miR-30c expression was negatively correlated with BCL9 expressions in CRC tissues. Furthermore, the findings of this study also showed that BCL9 was a direct target of miR-30c in CRC and miR-30c could inhibit the CRC proliferation by binding to its 3'-UTR. CONCLUSIONS: This study showed that miR-30c overexpression inhibited CRC proliferation via the regulation of BCL9, suggesting that miR-30c may be a new molecular therapeutic target for CRC.

Effects of miR-132 on proliferation and apoptosis of pancreatic cancer cells via Hedgehog signaling pathway.

OBJECTIVE: Micro ribonucleic acids (miRNAs) and Hedgehog (Hh) signaling pathway play key roles in the proliferation, migration and invasion of tumor cells. The aim of this study was to investigate the role of miR-132 and Hh signaling pathway in the proliferation and apoptosis of pancreatic cancer cells, and to investigate the possible underlying mechanism. MATERIALS AND METHODS: The expressions of miR-132 and Shh (a ligand of Hh) in clinical pancreatic cancer specimens and pancreatic cancer cell lines were determined by qRT-PCR. Meanwhile, the correlation between the two molecules was analyzed. Pancreatic cancer cell line (MiaPaCe-2a) was transfected with miR-132 mimics and inhibitor. The effects of miR-132 up- and down-regulation on the expressions of miR-132, Shh, Cyclin-D1, cleaved Caspase-3 and cleaved Caspase-9 were detected. In addition, the exact role of miR-132 in the proliferation, apoptosis and distribution of MiaPaCe-2a cells were investigated. RESULTS: The expression level of miR-132 in pancreatic cancer specimens and pancreatic cancer cell lines was significantly elevated when compared with that of control group. Meanwhile, miR-132 expression was negatively correlated with the expression level of Shh. Moreover, transfection with miR-132 mimics evidently up-regulated miR-132 expression. Moreover, miR-132 up-regulation significantly decreased the mRNA and protein expressions of Shh, facilitated the proliferation of MiaPaCe-2a cells, reduced the protein expressions of Cyclin-D1, cleaved Caspase-3 and cleaved Caspase-9, and suppressed cell apoptosis. On the contrary, miR-132 inhibitor transfection significantly inhibited the proliferative activity of MiaPaCe-2a cells, decreased the proportion of cells in G1 phase, and increased the proportion of cells in G2/M phase. CONCLUSIONS: MiR-132 promotes proliferation and inhibits apoptosis of pancreatic cancer cells through Hh signaling pathway.

MicroRNA-31 inhibits osteosarcoma cell proliferation, migration and invasion by targeting PIK3C2A.

OBJECTIVE: To elucidate the role of microRNA-31 (miR-31) in osteosarcoma and the molecular mechanism of miR-31 in the proliferation, migration, and invasion of osteosarcoma. PATIENTS AND METHODS: Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) was used to examine the expression of microRNA-31 in human osteosarcoma tissues. Pearson's chi-squared test was used to analyze the correlation between microRNA-31 and clinicopathological features. Proliferation, migration, invasion, and PI3K3C2A protein in treated osteosarcoma cells were detected by Cell Counting Kit-8 (CCK-8) assay, transwell assay without Matrigel, transwell assay with Matrigel, and Western blot analysis, respectively. RESULTS: qRT-PCR showed that miR-31 was down-regulated in osteosarcoma tissues compared with paired para-tumor bone tissues. The lower level of miR-31 was closely associated with high-grade osteosarcoma, metastasis, and poor overall survival. CCK-8 and transwell assay showed that miR-31 inhibited osteosarcoma cells proliferation, migration, and invasion. According to luciferase assay, miR-31 inhibits osteosarcoma cell proliferation, migration, and invasion through inhibiting PIK3C2A. Reversely, overexpression of PIK3C2A inhibited partial effect of miR-31 on proliferation, migration, and invasion in vitro. CONCLUSIONS: MiR-31 inhibits osteosarcoma cell proliferation, migration, and invasion by targeting PICK3C2A. MiR-31 can thus be used as a therapeutic target in osteosarcoma treatment.

Association of the upregulation of LncRNA00673 with poor prognosis for colorectal cancer.

OBJECTIVE: To investigate the effect and clinical significance of long non-coding RNA 00673 (lncRNA00673) in the occurrence and development of colorectal cancer (CRC) through the research on the expression level, biological effect and clinical significance of lncRNA00673 in CRC. PATIENTS AND METHODS: The relative expression of lncRNA00673 in 71 pairs of CRC tissues and cells was detected by quantitative Real-time polymerase chain reaction (qRT-PCR). The correlation of the relative expression of lncRNA00673 with the clinicopathological features of CRC patients was analyzed. The lncRNA00673 interference sequence was designed and synthesized, and its transfection efficiency was detected by qRT-PCR assays. 3-(4,5-Dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay and clone formation experiments were performed to investigate the effect of lncRNA00673 on the proliferation ability of CRC cells. RESULTS: In CRC tissues of 71 patients, there were 51 patients whose lncRNA00673 level was up-regulated compared with that of cancer-adjacent tissues. The highly expressed lncRNA00673 was positively correlated with tumor, node and metastasis (TNM) staging, regional lymph node metastasis, distant metastasis and tumor size in CRC patients. Cox proportional-hazards regression model showed that the highly expressed lncRNA00673 was an independent risk factor for the overall survival of CRC patients. Kaplan-Meier curve analysis showed that highly expressed lncRNA00673 was significantly associated with the relatively lower overall survival (OS). MTT and clone formation experiments showed that knockdown of lncRNA0673 could inhibit the proliferation of CRC cells. CONCLUSIONS: The expression level of lncRNA00673 is up-regulated in CRC tissues and cells, which is related to the degree of malignancy and poor prognosis. LncRNA00673 can be used as a potential molecular marker for the prognosis of CRC.

[Prevention for glucocorticoid-induced osteonecrosis of femoral head: a long-term clinical follow-up trail].

Objective: To evaluated the outcome of prevention and treatment for glucocorticoid-induced osteonecrosis of femoral head with anticoagulant and vasodilator drugs. Methods: A prospective, randomized, double-blind study was performed. From August 2003 to August 2006, 58 patients with large amounts of hormone therapy in the Zhongshan Hospital Affiliated Dalian University were enrolled and randomly assigned to the control group (placebo) or preventive group (anticoagulant and vasodilator drugs). And we prospectively analyzed the clinical outcomes of 24 patients with glucocorticoid-induced osteonecrosis of femoral head early stage (treatment group)treated by anticoagulant and vasodilator drugsat the same time. Disease incidence rate and progression were evaluated by radiography and magnetic resonance imaging (MRI), Follow-up of patients with femoral head survival curve was drawn. The Harris Hip Score and the Short Form 36 health survey were used to rate hip function and quality of life, respectively. Results: Thus, a total of 80 patients were assessed in this study, 24 cases in control group[follow up from 7.5 to 13.0(10.7±1.6)years], 22 cases in preventive group and 24 cases in treatment group. There was significant difference in theincidence rate of Osteonecrosis of femoral head, survive rate of femoral head and HHS score between the control groupand preventive group(41.7% vs 13.6%, 66.7% vs 70.8% , P<0.01). Conclusion: Anticoagulant and vasodilator drugs could effect on preventing theglucocorticoid-induced osteonecrosis of femoral head, reducing disease progression, or improving life quality.

Porous tantalum seeded with bone marrow mesenchymal stem cells attenuates steroid-associated osteonecrosis.

OBJECTIVE: Bone marrow mesenchymal stem cells (BMMSCs) have been widely applied in osteonecrosis. However, lack of biomechanical support limited application of BMMSCs. And porous tantalum (PTA) has been identified as a cell-friendly scaffold for bone regeneration. Herein, we aimed to investigate the efficacy of PTA seeded with BMMSCs in the treatment of osteonecrosis. MATERIALS AND METHODS: After the production of PTA seeded with BMMSCs, MTT and GFP were performed to identify the proliferation and adhesion of BMMSCs respectively, which was further examined by scanning electron microscopy (SEM). And real-time PCR was also used to determine mRNA level of osteogenic markers, including Alp, OCN, OPN, Col I and Runx-2 in BMMSCs. Nineteen adult rabbits were applied for building steroid-associated osteonecrosis (SAON) models. Bone formation rate (BFR) and mineral apposition rate (MAR) were determined. And Goldner Trichrome Staining was used in these SAON models, which further confirmed the efficacy of PTA seeded with BMMSCs in SAON. RESULTS: PTA seeded with BMMSCs showed excellent biocompatibility. Additionally, SEM assay showed that BMMSCs adhered tightly and spread fully in the pores of PTA. Next, the expression of ALP and OPN mRNA in BMMSCs were significantly (p < 0.05) higher in the PTA-treated group compared to those in the PTA-untreated group. Furthermore, compared to those treated by only PTA, the dynamic bone formation in rabbits treated by PTA seeded with BMMSCs was significantly increased (p < 0.001) at both week 3rd and week 6th. CONCLUSIONS: The product, PTA seeded with BMMSCs, was successfully produced, and was determined as high efficacy for treatment of steroid-associated osteonecrosis. PTA seeded with BMMSCs may afford a promising option for treating osteonecrosis.

Study of human acellular amniotic membrane loading bone marrow mesenchymal stem cells in repair of articular cartilage defect in rabbits.

The aim of this study was to investigate the repair effect of human acellular amniotic membrane (HAAM) loading bone marrow mesenchymal stem cells (BMSCs) on articular cartilage defect in rabbits. Rabbit BMSCs were isolated and cultured, and they were then inoculated on HAAM to prepare the complex of HAAM and BMSCs. Twenty-four rabbits were randomly divided into groups A and B, with 12 animals in each group. The left and right sides were used as the experimental and control sides, respectively. The models of bilateral articular cartilage defect were established. The defect areas on the experimental side in groups A and B were implanted with the complex of HAAM and BMSCs and HAAM alone, respectively. The control sides of the two groups were not implanted with any material. In the 8th and 12th week after surgery, gross observation, histological examination and cartilage defect scoring were performed. In the 8th and 12th postoperative week, gross observation and histological observation showed that dense cartilage-like cells appeared in group A but not in group B, indicating preferable cartilage repair. The cartilage defect score on the experimental side in group A was 5.31 ± 0.68 in the 8th week and 3.23 ± 0.52 in the 12th week, and that in group A was significantly lower than in group B (P < 0.05). HAAM loading BMSCs has a good repair effect on articular cartilage defect under an in vitro environment.

Association of the g.27563G>A osteoprotegerin genetic polymorphism with bone mineral density in Chinese women.

Osteoporosis is a common multifactorial disease in postmenopausal women. This study aimed to investigate the association of the g.27563G>A osteoprotegerin (OPG) genetic polymorphism with bone mineral density (BMD) and osteoporosis. A case-control study was carried out with 435 osteoporosis postmenopausal women cases and 442 age-matched healthy controls. The BMD at the femoral neck hip, lumbar spine (L2₋4), and total hip were assessed by Norland XR-46 dual-energy X-ray absorptiometry. The genotypes of the g.27563G>A genetic polymorphism were detected by created restriction site-polymerase chain reaction and verified by DNA sequencing methods. We detected that the g.27563G>A genetic polymorphism was a non-synonymous mutation that resulted in an arginine (Arg) to glutamine (Gln) amino acid replacement (p.Arg333Gln). Significant differences were found in the BMD of the femoral neck hip, lumbar spine (L2₋4), and total hip among different genotypes of the g.27563G>A genetic polymorphism. Subjects with the genotype GG had significantly higher BMD values than those with genotypes GA and AA (P < 0.05). Our data indicated that the A allele of the g.27563G>A genetic polymorphism in OPG could be associated with lower BMD values in the Chinese postmenopausal women evaluated, and that it might be an increased risk factor for osteoporosis.

Atmospheric deposition, mineralization and leaching of nitrogen in subtropical forested catchments, South China.

In recent years, China has conducted considerable research focusing on the emission and effects of sulphur (S) on human health and ecosystems. By contrast, there has been little emphasis on anthropogenic nitrogen (N) so far, even though studies conducted abroad indicate that long-range atmospheric transport of N and ecological effects (e.g. acidification of soil and water) may be significant. The Sino-Norwegian project IMPACTS, launched in 1999, has established monitoring sites at five forest ecosystems in the southern part of PR China to collect comprehensive data on air quality, acidification status and ecological effects. Here we present initial results about N dynamics at two of the IMPACTS sites located near Chongqing and Changsha, including estimation of atmospheric deposition fluxes of NOx and NHx and soil N transformations. Nitrogen deposition is high at both sites when compared with values from Europe and North America (25-38 kg ha(-1) yr(-1)). About 70% of the deposited N comes as NH4, probably derived from agriculture. Leaching of N from soils is high and nearly all as NO3-. Transformation of N to NO3- in soils results in acidification rates that are high compared to rates found elsewhere. Despite considerable leaching of NO3- from the root zone of the soils, little NO3- appears in streamwater. This indicates that N retention or denitrification, both causing acid neutralization, may be important and probably occur in the groundwater and groundwater discharge zones. The soil flux density of mineral N, which is the sum of N deposition and N mineralization, and which is dominated by the N mineralization flux, may be a good indicator for leaching of NO3- in soils. However, this indicator seems site specific probably due to differences in land-use history and current N requirement.

[Observation on sex-linkage inheritance of skin color in some local breeds of chicken].

The sex-linkage inheritance of skin color was observed in some local breeds of chicken. The results showed that there was a pair of melanin genes PP in autosomes in Taihe Silky Fowl, an Id gene in sex chromosome in Xianju Fowl, Xiaoshan Fowl and Beijing You Fowl, and an id gene in sex chromosome in Langshan Fowl, Gushi Fowl and Taihe Silky Fowl, respectively. The offspring produced by crossing of Taihe Silky Fowl(Male) with the fowl(Female) carrying Id gene (such as Xianju Fowl, Xiaoshan Fowl and Beijing You Fowl) can autosexingly determinate by skin color, male chicken has yellow skin, female chicken has black skin.

[Treatment of ischemic necrosis of femoral head by the transfer of vascular pedicled iliac periosteum].

OBJECTIVE: To investigate the therapeutical effect of treatment of ischemic necrosis of femoral head by the transfer of vascular pedicled iliac periosteum. METHODS: From June 1983 to August 1997, 106 cases with ischemic necrosis of femoral head (II stage in 64 cases, III stage in 39 cases, IV stage in 3 cases) were treated by the transfer of vascular pedicled iliac periosteum with ascending branch of lateral femoral circumflex vessel or deep circumflex iliac vessel pedicle. RESULTS: Followed up 2 years and 4 months to 16 years, there were excellent in 54 cases, better in 38 cases, moderate in 9 cases, poor in 5 cases, and 86.8% in excellent rate according to the criterion of the therapeutical effect on the repair and reconstruction of adult ischemic necrosis of femoral head. CONCLUSION: Treating ischemic necrosis of femoral head by the transfer of vascular pedicled iliac periosteum has the advantage of constant pedicle, easily drawing materials and reliable therapeutical effect.

Copper/zinc superoxide dismutase, nuclear DNA content, and progression in human gliomas.

To our knowledge, there have been no previous reports regarding the immunohistochemistry and image cytometry to demonstrate elevated Copper/zinc superoxide dismutase (Cu/Zn SOD) expression and numbers of the clonal cells in human gliomas. In 30 well-studied patients with gliomas, immunoreactivity for Cu/Zn SOD and cytometric evidence of DNA ploidy in the G2M cell cycle phase were evaluated from routinely prepared tissue blocks. Cu/Zn SOD positive tumor cells were shown in 8 of 13 glioblastomas (mean quantitative immunoreactivity SOD score; 1), 3 of 8 anaplastic gliomas (score; 0.6), and none of 9 low-grade gliomas. The differences in SOD score was not significant. In hypertetraploid glioblastomas, time to progression was shorter than for hypertetraploid of anaplastic gliomas, while SOD scores were not significantly different. The same relationship held for tetraploid specimens. Considering variables in combination, hypertetraploid gliomas with high SOD immunoreactivity showed a significantly short time to progression (p < 0.05) (1-5 months after radiotherapy and chemotherapy) compared with hypertetraploid, low-SOD immunoreactivity gliomas or tetraploid, low-SOD immunoreactivity gliomas. The tumor cells with high SOD activity also tended to be resistant for radiotherapy and anticancer drugs. Those results were suggested that the high grade glioma with a single clone and low SOD activity were effective for radiotherapy associated with oxidative stress, and that the high grade gliomas with more than two clones and high SOD activity were very less effective for same therapy. Cu/Zn SOD activity and the degree of the clonality in human gliomas should be very important factors influencing a choice of oxidative cytotoxic treatment.

[Therapeutic effect of hyperbaric oxygen on recovery of surgically repaired peripheral nerve injury].

114 patients with peripheral nerve injury were treated microsurgically. 54 patients with 65 nerve injuries were given hyperbaric oxygen as supplementary treatment. The excellent and good results of the 65 nerves accounted for 89.2%, but of those nerves in patients not treated with hyperbaric oxygen for 73.2% (P less than 0.05). The result was better in the hyperbaric oxygen group regardless of the repairing method used, particularly in those nerve grafting had been done (24.9, P less than 0.05). As for the time lapsed from injury to surgery. The results of fresh injuries showed no difference in either group, but hyperbaric oxygen was of much benefit to the old cases: the longer the delay the greater in difference between different groups, but better results were obtained in the hyperbaric oxygen group. The early the operation the better the result in either group.