[Analysis of clinicopathological features and prognosis of sporadic synchronous multiple primary colorectal cancers].

Objective: To investigate the impact of relative locations of multiple foci and microsatellite status of sporadic, synchronous, multiple, primary, colorectal carcinomas on clinicopathological features and prognosis. Methods: The clinicopathologic and prognostic data of 278 patients with sporadic, synchronous, multiple, primary, colorectal carcinomas who had been admitted to the Department of Colorectal Surgery at Zhejiang Cancer Hospital from January 2008 to July 2022 were retrospectively collected. The patients were categorized into three groups based on the relative locations of their multiple cancer foci: (1) a right-sided group that comprised patients with multiple cancer foci in the cecum, ascending colon, hepatic flexure of the colon, and transverse colon; (2) a left-sided group that comprised patients with multiple cancer foci in the splenic flexure of the colon, descending colon, sigmoid colon, and rectum; and (3) a left- and right-sided group that comprised patients with multiple cancer foci in the right half of the colon and left half of the colon/rectum. Additionally, the patients were further divided into two groups based on microsatellite status: a high microsatellite instability (MSI-H) and a low MSI/stable MSI (MSI/L&MSS) group. We compared differences in clinical characteristics and prognostic indicators between these groups. The χ2 test was utilized to compare selected clinical characteristics, whereas Kaplan-Meier survival analyses and log-rank tests were performed to compare their effects on prognosis. Result: Among 278 patients with SSCRC, 256 (92.1%) presented with two cancer foci and 22 (7.9%) with more than two foci. Additionally, 255 patients (91.7%) had adenocarcinomas, whereas the remaining 23 (8.3%) had mucinous adenocarcinomas. Lymph node metastases were identified in 136 patients (48.9%); the cancer foci had infiltrated beyond the muscular layer in 238 (85.6%); and 147 patients (52.9%) were diagnosed with TNM Stage III-IV disease. There were 155 patients (55.8%) in the left-sided group, 55 (19.8%) in the right-sided group, and 68 (24.5%) in the left- and right-sided group. Immunohistochemical examination of all four mismatch repair proteins were performed in 199 cases, revealing that 166 of these patients had MSI/L&MSS and 33 MSI-H disease. In the left-sided, left- and right-sided, and right-sided groups, the proportion of women was 16.8% (26/155), 26.5% (18/68), and 49.1% (27/55), respectively; these differences are statistically significant (χ2=22.335, P<0.001). The proportions of patients with more than three cancer foci were 5.2% (8/155), 16.2% (11/68), and 5.5% (3/55), respectively; these differences are statistically significant (χ2=8.438, P=0.015). The proportions of mucinous adenocarcinomas were 4.5% (7/155), 8.8% (6/68), and 18.2% (10/55), respectively; these differences are statistically significant (χ2=10.026, P=0.007). The proportions of patients with lymph node metastases were 55.5% (86/155), 48.5% (33/68), and 30.9% (17/55); these differences are statistically significant (χ2=9.817, P=0.007). The proportions of patients with Stage T3 & T4 disease in each group according to location were 81.3% (126/155), 88.2% (60/68), and 94.5% (52/55), respectively; these differences are statistically significant (χ2=6.293,P=0.043). The proportions of TNM Stage III-IV tumors were 59.4% (92/155), 54.4% (37/68), and 32.7% (18/55), respectively; these differences are statistically significant (χ2=11.637, P=0.003). Age, size of cancer foci, presence of distant metastasis, adenoma, nerve invasion, and vascular invasion did not differ significantly between the three groups (all P>0.05). Compared with those with MSI-H, patients with MSI/L&MSS disease were more likely to be aged >65 years and male (50.6% [84/166] vs. 15.2% [5/33], χ2=13.994,P<0.001; 80.7% [134/166] vs. 54.5% [18/33], χ2=10.457,P=0.001), more likely to be in the left-sided group (63.3% [105/166] vs. 24.2% [8/33], χ2=18.232, P<0.001), had a higher proportion of cancer foci of diameter <4 cm (54.8% [91/166] vs. 33.3% [11/33], χ2=5.086,P=0.024), and a lower proportion of mucinous adenocarcinomas (4.2% [7/166] vs. 27.3% [9/33], χ2=19.791,P<0.001), more likely to develop distant metastases (22.3% [37/166] vs. 6.1% [2/33], χ2=4.601,P=0.032), more likely to have lymph node metastases (57.2% [95/166) vs. 24.2% [8/33], χ2=11.996,P<0.001) and nerve invasion (28.9% [48/166] vs. 6.1% [2/33], χ2=7.643, P=0.006), had a higher proportion of TNM Stage III-IV disease (60.2% [100/166] vs. 24.2% [8/33], χ2=14.374, P<0.001), and a smaller proportion of family history of tumors (28.9% [48/166] vs. 60.6% [20/33], χ2=12.228, P<0.001). All the above-listed differences are statistically significant (all P<0.05). The differences in number of cancer foci, depth of infiltration, presence or absence of adenomas, and vascular invasion were not statistically significant (all P>0.05). In the 33 patients with MSI-H status and mismatch repair protein loss, the highest frequency of deletion was found in PMS-2 (66.7%, 22/33), followed by MLH-1 (57.6%, 19/33), whereas the proportions of MSH-2 (33.3%, 11/33) and MSH-6 (24.2%, 8/33) deletions were relatively low. There were statistically significant differences in the 3-year overall survival rates among the groups according to relative locations of cancer foci. The 3-year overall survival rates were 96.8%, 79.6%, and 88.5% in the right-sided, left- and right-sided, and left-sided groups, respectively (P=0.021). As to microsatellite status, the 3-year overall survival rate of patients with MSI-H disease was 93.8%, which is significantly better than the 78.4% for those with MSI/L & MSS (P=0.026). Conclusions: Among sporadic, synchronous, multiple, primary, colorectal carcinomas, those with right-sided disease had the deepest local infiltration, whereas those with left-sided disease had the greatest number of lymph node metastases, most advanced clinical TNM stage, lowest percentage of MSI-H disease, and the poorest prognosis.

The choice of spectral similarity algorithms influences suspected soil sample provenance.

Similarity algorithms are commonly used in soil forensic applications to help identify similar samples from an existing reference library as possible source locations of unknown target samples. These algorithms are well-suited to compare soil spectra. However, different similarity algorithms may lead to different clusters of similar samples, and thus different strengths of evidence in forensic investigations. To quantify this, we conducted a study to evaluate the influence of seven similarity algorithms on soil provenance, using as a sample set a soil spectral library consisting of 280 soil profiles from Anhui Province, China. This library includes three spatial scales of datasets: provincial (DSp), county (DSc) and field (DSf). A set of ten samples covering a wide range of spectra variations were selected from the DSf dataset as the "unknown" samples, with the remaining being used as the reference samples. This study aimed to: (1) evaluate how several commonly-used similarity algorithms, namely Euclidean distance (ED), Mahalanobis distance (MD), Spectral angle mapper (SAM), and Spectral information divergence (SID), as well as variants of several of these measured in standardized principal component space computed from the spectra (ED_PCA, MD_PCA and SAM_PCA), influence the identification of the matched similar samples; (2) determine the overlap in sample selection between different similarity algorithms; (3) propose best practices for similarity algorithms applied to soil forensic analysis using spectroscopy. The use of different similarity algorithms did influence the selection of most similar samples. The similarity algorithms calculated in PC space (ED_PCA, MD_PCA and SAM_PCA) performed slightly better than their counterparts calculated in spectral space. Due to the availability of a detailed spectral library, regardless of the different similarity algorithms used, the matched most similar samples were all located close to the unknowns, mostly within 3 km, with one exception. That is, the varied choices of different similarity algorithms hardly influenced the conclusion of soil provenance in this case. In general, MD_PCA, SAM and ED were the best similarity algorithms overall. However, since there was no single best algorithms for all cases, we recommend the joint use of MD_PCA, SAM and ED as an ensemble. Indications of possible sample provenance from these similarity measured can be useful evidence to complement evidence from other methods in a forensic investigation.

[The correlation between methylation in HPV16 long control region and cervical intraepithelial neoplasia grade 2 or more: a Meta-analysis].

Objective: To investigate the correlation between methylation in human papillomavirus 16 (HPV16) long control region (LCR) and cervical intraepithelial neoplasia grade ≥2 (CIN2+). Methods: The literature retrieval was conducted by using the databases of PubMed, Embase, Cochrane Library, Web of Science, CNKI, Wanfang data and Weipu according to the inclusion and exclusion criteria, and the retrieval period was from the establishment of the databases to February 27th, 2022. Software RevMan 5.3 and Stata 15.1 were used for Meta-analysis. Results: A total of 17 literatures were included involving 1 421 subjects. Results of Meta-analysis showed that OR of the correlation between methylation of HPV16 LCR and CIN2+ was 1.56 (95%CI: 0.70-3.47). Subgroup analysis showed that methylation of the 5' terminal, enhancer and promoter regions were not associated with CIN2+, while in four E2 binding sites (E2BS), the methylation of E2BS1, E2BS3 and E2BS4 increased the risk of CIN2+, with the ORs of 3.92 (95%CI: 1.92-7.99), 10.50 (95%CI: 3.67-30.04) and 3.65 (95%CI: 1.58-8.41), respectively. However, subgroup analysis on E2BS2 was not performed due to the limitation of the number of literatures. According to the different sources of population, the risk of CIN2+ in Chinese population was associated with methylation of HPV16 LCR (OR=2.14, 95%CI: 1.31-3.50). There was a correlation between the risk of CIN2+ and HPV16 LCR methylation in the population with pyrosequencing of HPV16 LCR, and OR was 1.75 (95%CI: 1.03-2.98). Conclusion: The risk of CIN2+ is correlated with the methylation of E2BS in HPV16 LCR, which can be used as potential biomarkers.

[Progress in research of genomic epidemiology of pathogenic fungi].

Genomic epidemiology, based on whole-genome sequencing technology and bioinformatics analysis, can make up for the shortcomings of traditional molecular typing methods and provide a novel insight for the genetic evolution and transmission of pathogenic fungi. The combination of genetic information and epidemiological methods of pathogenic fungi can predict fungi transmission routes and risks, and provide a theoretical basis for the development of public health strategies for fungi infection prevention and control. This paper summarizes the development of molecular epidemiology and genomic epidemiology, as well as the application of genomic epidemiology methods in the analyses of genetic relationship, origin, evolution, drug resistance, virulence, and genome-wide association of pathogenic fungi, and discusses the development of pathogenic fungi genomic epidemiology.

[CT-based morphological and hemodynamics analysis for rupture risk of mirror intracranial aneurysm].

Objective: To compare the morphological and hemodynamic features of mirror intracranial aneurysms (MIAs) on CT angiography (CTA), and to elucidate the rupture risk factors of MIAs. Methods: This study retrospectively collected 29 patients with 58 digital subtraction angiography (DSA) or surgically confirmed MIAs from January 2010 to December 2016 in Jinling Hospital, Medical School of Nanjing University. Among them, there are 6 males and 23 females, aged from 40 to 83 (61±11) years old. Based on the results of hemorrhagic manifestation, 58 MIAs were divided as the ruptured (n=29) group and unruptured group (n=29). In addition, according to the location of aneurysms, they were further divided into the subgroup of posterior communicating MIAs (n=32) and non-posterior communicating MIAs (n=26). Clinical data of the patients and the morphological parameters of the MIAs were collected. Computational fluid dynamics (CFD) analysis was performed to obtain hemodynamic parameters, such as pressure (P), wall shear stress (WSS), wall shear stress gradient (WSSG), and oscillatory shear index (OSI). The coefficient of variation (CV) was used to describe the aforementioned hemodynamic parameters of intracranial aneurysms, so the index after CV adjustment is expressed as PCV , WSSCV , WSSGCV , OSICV . Characteristics between ruptured and unruptured groups were compared. Conditional logistic regression analysis was conducted to evaluate the rupture risk factors of MIAs. Results: Among the 29 pairs of mirror aneurysms, 16 pairs were distributed in bilateral posterior communicating arteries (55%), 9 pairs distributed in bilateral middle cerebral arteries (31%), and 4 pairs distributed in bilateral internal carotid arteries (14%). Compared with the unruptured MIAs group, the ruptured aneurysms group usually had a larger maximum diameter, neck width, and size ratio (SR) [4.98 (3.18, 6.79) mm vs 3.20 (2.10, 4.31) mm, 4.19 (3.46, 5.95) mm vs 4.05 (3.23, 5.02) mm, 1.69 (0.81, 2.28) vs 0.96 (0.67, 1.49)] (all P<0.05). In the subgroup hemodynamic analysis of MIAs, the ruptured aneurysms had higher WSSCV and WSSGCV than the contralateral unruptured ones [1.00(0.87, 1.21) vs 0.65(0.57, 0.87), 1.09(0.56, 1.90) vs 0.57(0.50, 1.13), 1.52 (1.34, 1.80) vs 1.21 (1.07, 1.38), 1.52±0.46 vs 1.21±0.23] (all P<0.05), while the PCV was lower than the contralateral unruptured ones [0.004 (0.002, 0.008) vs 0.010 (0.006, 0.013), 0.003 (0.002, 0.011) vs 0.009 (0.002, 0.066)] (both P<0.05). Logistic regression analysis showed that high WSSGCV was an independent risk factor for MIAs rupture (OR=279.20(95%CI:1.10-71 028.28)). Conclusion: The maximum diameter, neck width, and SR were considered as a reliable morphological parameters to distinguish the ruptured status of MIAs, higher WSSGCV in the aneurysm sac are highly correlated with MIAs rupture.

Application of principal component analysis with instrumental analysis and sensory evaluation for assessment of chicken breast meat juiciness.

1. The objectives of this study were to use principal component analysis (PCA) to analyse the variability of the three instrumental and 14 descriptive sensory properties of chicken breast meat. The meat was cooked until the internal temperature reached 85°C and further cooked for 0, 20, and 40 min. The second objective was to identify the most critical variables for assessing meat juiciness.2. Cooking loss and moisture content exhibited high correlation with sensorial moisture release and mouth feel.3. The distribution of objects on the axes of the first two principal components (PCs) enabled the identification of three groups undergoing different cooking durations. The four major PCs explained 80.0% of the total variability.4. Cooking loss, moisture content, water-holding capacity, sensorial moisture release and mouth feel were demonstrated as the most effective variables for the first two PCs. PCA with instrumental and sensory analyses proved an effective procedure for systematically and comprehensively judging chicken meat juiciness.

How similar is "similar," or what is the best measure of soil spectral and physiochemical similarity?

Spectral similarity indices were used to select similar soil samples from a spectral library and improve the predictive accuracy of target samples. There are many similarity indices available, and precisely how to select the optimum index has become a critical question. Five similarity indices were evaluated: Spectral angle mapper (SAM), Euclidean distance (ED), Mahalanobis distance (MD), SAM_pca and ED_pca in the space of principal components applied to a global soil spectral library. The accordance between spectral and compositional similarity was used to select the optimum index. Then the optimum index was evaluated if it can maintain the greatest predictive accuracy when selecting similar samples from a spectral library for the prediction of a target sample using a partial least squares regression (PLSR) model. The evaluated physiochemical properties were: soil organic carbon, pH, cation exchange capacity (CEC), clay, silt, and sand content. SAM and SAM_pca selected samples were closer in composition compared to the target samples. Based on similar samples selected using these two indices, PLSR models achieved the highest predictive accuracy for all soil properties, save for CEC. This validates the hypothesis that the accordance information between spectral and compositional similarity can help select the appropriate similarity index when selecting similar samples from a spectral library for prediction.

Forensic soil source identification: comparing matching by color, vis-NIR spectroscopy and easily-measured physio-chemical properties.

This study evaluates to what degree soil samples associated in characteristic space are also close in geographical space, i.e., the possible location from which an unknown sample was obtained in a forensic investigation. The study compares similarity computed from Munsell colors, RGB colors, and full visible-near infrared (vis-NIR) spectra by the spectral angle mapper with similarity based on six easily-measured physio-chemical properties. The reference area is Anhui Province, China with three scales of datasets: provincial, county, and field. Ten diverse "unknown" samples were selected by the Kennard-Stone algorithm from the field-scale dataset and their matches in characteristic space from the several datasets were found by the different methods. The geographic distances of the matches to the "unknowns" were used to evaluate the source identification ability. When a detailed library with local samples is present, a limited set of physio-chemical properties achieved higher geographic accuracy than the color and spectral methods. However, with a regional library the spectral and color methods are superior. Distances in RGB space reveal finer differences than exact matching in Munsell space, but whole-spectra matching outperforms both, because of the rich information influenced by more soil properties than influencing color. We recommend the use of soil vis-NIR spectra as a priority indicator for forensic soil analysis because of its success in this study and its ability to work non-destructively on small quantities of soil.

MiR-136 triggers apoptosis in human gastric cancer cells by targeting AEG-1 and BCL2.

OBJECTIVE: Gastric cancer is the second most prevalent cancer across the globe and accounts for about 10% of new cancer cases. It is one among the leading causes of cancer-related deaths around the world. Recently, microRNAs have been identified as important therapeutic targets for the treatment of several cancers owing to their potential to target multiple genes and hinder several biological processes such as proliferation and apoptosis. In the current study, we investigated the potential of miR-136 as therapeutic target for gastric cancer. MATERIALS AND METHODS: Total RNA was isolated by RNeasy RNA isolation kit and cDNA was prepared byRevertAid cDNA synthesis kit. The transcript analysis was carried out by quantitative RT-PCR. The transfection of miR-136 mimics or plasmids was carried out by using the Lipofectamine 2000 reagent. Apoptosis was detected by DAPI and acridine orange/ethidium bromide (AO/EB) staining. Protein expression was examined by Western blotting. RESULTS: The results indicated that the expression of miR-136 is significantly downregulated in gastric cancer cells. Transfection and subsequent overexpression of miR-136 in gastric cancer cells significantly promoted apoptosis as evident from DAPI and OA staining. In silico analysis revealed AEG-1 and BCL2 to be the potential targets of miR-136. Therefore, the expression of AEG-1 and BCL2 was determined in untreated control, cisplatin treated control and miR-136 transfected AGS gastric cancer cells. The results revealed that overexpression of miR-136 expression causes significant downregulation of AEG-1 and BCL2 protein expression. CONCLUSIONS: Taken together, we conclude that miR-136 promotes apoptosis in gastric cancer cells by targeting AEG-1 and BCL2. Therefore, miR-136 may prove as a potential therapeutic target for the treatment of gastric cancer.

Diode-wing-pumped electro-optically Q-switched 2 µm laser with pulse energy scaling over ten millijoules.

Characteristics of diode-wing-pumped highly efficient Tm:LuAG lasers running both in continuous wave (CW) and electro-optical Q-switching regimes have been investigated. Using a simple plane-plane cavity, a maximum CW output power of 8.5 W has been achieved with a corresponding slope efficiency of 44.5% by "wing pumping" at 790 nm. With a V-shaped cavity, a diode-wing-pumped MgO:LiNbO3 crystal based electro-optically Q-switched Tm:LuAG laser at 2022.9 nm delivered a maximum pulse energy of 10.8 mJ and a minimum pulse width of 52 ns at a corresponding repetition rate of 100 Hz. To the best of our knowledge, the achieved CW output power and Q-switched pulse energy have both set records for all-solid-state Tm:LuAG lasers, which well reveals an efficient way to generate high-power and high-energy lasers at 2 µm wavelength.

[Preoperative dynamic contrast-enhanced MRI can reduce the rate of tumor-positive resection margins after breast conserving surgery in patients with early non-mass breast carcinoma].

Objective: To investigate the value of preoperative dynamic contrast-enhanced MRI in reducing the rate of tumor-positive resection margins after breast conserving surgery in patients with early non-mass breast carcinoma. Methods: Seventy-two patients with early non-mass breast carcinoma received ultrasonographic and mammographic examination and subsequently underwent dynamic contrast-enhanced MRI examination before breast conserving surgery. The control group consisted of 74 patients who had early non-mass breast carcinoma. They only received ultrasonographic and mammographic examination and didn't undergo contrast-enhanced MRI examination. The comparison of the rate of tumor-positive resection margins between two groups was performed. The MRI findings that had the significant influence on the rate of tumor-positive resection margins were analyzed using Logistic regression model. Results: In 28 patients (28/72, 38.9%), dynamic contrast-enhanced MRI could correct or supplement the ultrasonographic and mammographic findings and resulted in the reasonable change of surgical program. The preoperative MRI examination group (n=30) had lower rate of tumor-positive resection margins than control group for invasive ductal carcinoma (23.3% vs 40.0%, P=0.02), but there was no significant difference (21.4% vs 26.9%, P=0.10) between two groups for ductal carcinoma in situ (n=28). The preoperative MRI examination group (n=14) had lower rate of tumor-positive resection margins than control group for the other pathologic types of breast carcinoma (14.3% vs 38.9%, P=0.02). The statistical analysis on the basis of Logistic regression model showed that some main MRI findings, including change surrounding the tumor, distance between tumor and nipple and tumor size, had the significant influence on the rate of tumor-positive resection margins. Conclusion: Preoperative dynamic contrast-enhanced MRI significantly increased the accuracy of resection margins evaluation, and greatly reduced the rate of tumor-positive resection margins after breast conserving surgery in patients with early non-mass breast carcinoma.

Role of microRNAs in the treatment of type 2 diabetes mellitus with Roux-en-Y gastric bypass.

The aim of this study was to investigate the effect of Roux-en-Y gastric bypass (RYGB) on the peripheral blood microRNAs (miRNAs) of patients with type 2 diabetes mellitus (T2DM). miRNAs are small 20- to 22-nucleotide (nt) noncoding RNAs. They constitute a novel class of gene regulators that negatively regulate gene expression at the post-transcriptional level. miRNAs play an important role in several biological processes. Twelve patients with T2DM who were scheduled to undergo laparoscopic RYGB surgery were separated into two groups, using a body mass index of 30 kg/m2 as a cut-off point. Venous blood was collected before operation and 12 months after operation. A significant change was observed in the peripheral blood miRNA expression profile of both groups after RYGB surgery compared with those before operation. The expression levels of hsa-miR-29a-3p, hsa-miR-122-5p, hsa-miR-124-3p, and hsa-miR-320a were downregulated. The methylation state of the CpG sites within an approximately 400-bp genomic DNA fragment of each of the four miRNA genes, including about 200 bp upstream and 100 bp downstream of the pre-miRNA, did not vary after RYGB surgery. With remission of T2DM in both groups, RYGB could modulate the expression level of many peripheral blood miRNAs associated with lipid metabolism, insulin secretion, beta-cell function, and insulin resistance. The expression level of peripheral blood diabetes-related miRNA varied in patients with T2DM after receiving RYGB surgery, laying a strong foundation for future studies on this subject. The molecular mechanisms underlying RYGB surgery that can cause aberrant expression of miRNA remains to be determined.

Composite artificial semi-knee joint system.

OBJECTIVES: The purpose of the study was to investigate the clinical implantation protocol of custom-made artificial semi-knee joint based on computer-aided design so as to improve the limb salvage efficiency. MATERIALS AND METHODS: The custom-made artificial semi-knee joint was designed and manufactured based on rapid prototyping technology. The repeated modifications were carried out in the design and manufacture of the semi-knee joint, together with the operation protocol. Clinical trial was conducted on 2 cases of osteosarcoma, one receiving allograft prosthesis composite transplantation, and the other receiving synthetic bone graft prosthesis composite transplantation. The clinical outcomes of the 2 patients were evaluated. RESULTS: The custom-made artificial semi-knee joint met the clinical customization needs. In clinical trial, 18-month follow-up demonstrated the satisfactory knee joint function recovery in near future. CONCLUSIONS: The custom-made artificial semi-knee joint based on computer-aided design can afford satisfactory knee joint function recovery following allograft bone transplantation.

Diode-pumped continuous wave tunable and graphene Q-switched Tm:LSO lasers.

We have investigated the lasing characteristics of Tm:LSO crystal in three operation regimes: continuous wave (CW), wavelength tunable and passive Q-switching based on graphene. In CW regime, a maximum output power of 0.65 W at 2054.9 nm with a slope efficiency of 21% was achieved. With a quartz plate, a broad wavelength tunable range of 145 nm was obtained, corresponding to a FWHM of 100 nm. By using a graphene saturable absorber mirror, the passively Q-switched Tm:LSO laser produced pulses with duration of 7.8 µs at 2030.8 nm under a repetition rate of 7.6 kHz, corresponding to pulse energy of 14.0 µJ.

Integration of seawater and grey water reuse to maximize alternative water resource for coastal areas: the case of the Hong Kong International Airport.

Development, population growth and climate change have pressurized water stress in the world. Being an urbanized coastal city, Hong Kong has adopted a dual water supply system since the 1950s for seawater toilet flushing for 80% of its 7 million inhabitants. Despite its success in saving 750,000 m(3)/day of freshwater, the saline sewage (consisting of about 20-30% of seawater) appears to have sacrificed the urban water cycle in terms of wastewater reuse and recycling. Can seawater toilet flushing be applied without affecting the urban water cycle with respect to sustainable water resource management? To address this issue, we examined the entire urban water cycle and developed an innovative water resource management system by integrating freshwater, seawater and reclaimed grey water into a sustainable, low-freshwater demand, low-energy consumption, and low-cost triple water supply (TWS) system. The applicability of this novel system has been demonstrated at the Hong Kong International Airport which reduced 52% of its freshwater demand.

Establishing a new electrical conduction pathway by anastomosis of the right auricle and right ventricle assisted by mesenchymal stem cells in a canine model.

BACKGROUND: Electronic pacemakers are the primary treatment of complete atrioventricular (AV) block, but their use is associated with many complications. The aim of the present study was to create an alternative treatment for these patients. MATERIALS AND METHODS: Mesenchymal stem cells (MSCs) isolated from the bone marrow of a 3-month-old dog were cultured in vitro. The MSCs were labeled with 4', 6-diamidino-2-phenylindole (DAPI) before transplantation. We anastomosed the right auricle and right ventricle in 24 dogs, and transplanted labelled MSCs into the anastomotic area of 8 dog hearts. Using immunostaining we assessed survival and differentiation of the implanted cells at 8 weeks posttransplantation. Electrocardiography confirmed the secondary electrical conduction pathway. RESULTS: The ventricular current was captured by the electronic pacemaker in 21 dogs. Compared with the control group (surgery alone), pacemaker stimulus current was significantly less in the MSC group (surgery+MSCs). CONCLUSIONS: Anastomosis of the right auricle and right ventricle assisted by MSCs may be a new treatment for patients with complete AV block in the future.

Anti-angiogenesis by lentivirus-mediated small interfering RNA silencing of angiopoietin-2 gene in pancreatic carcinoma.

Angiopoietin-2 (Ang2) has been shown highly expressed in resected human pancreatic carcinoma samples, but the role of it is less clear. We were, therefore, interested in exploring the effects of Ang2 silencing on the angiogenesis and growth of pancreatic carcinoma. Lentivirus mediated Ang2 small hairpin RNA (LV-RNAi) were transfected into pancreatic carcinoma cell line MIA PaCa-2. Three groups were designed in this study: the control group (Mia PaCa-2 cells), the LV-NC group (cells transfected with the control GFP-lentivirus) and the LV-RNAi group (cells transfected with LV-RNAi). The mRNA and protein level of Ang2 gene were detected by real-time polymerase chain reaction and Western blot respectively. MTT assay and Flow Cytometry were used to detect the cell growth and apoptosis. Anti-angiogenesis effect was measured by chick embryo chorioallantoic membrane (CAM) assay. In nude mice bearing tumors, after treatment with intratumoral injection of LV-RNAi, mice growth and tumor volume were observed, and the expression of Ang2, VEGF and CD34 were measured by immunohistochemistry. Compared with the control group and the LV-NC group, the mRNA and protein level of Ang2 gene were successfully knocked down in LV- RNAi group. Also the vessel count was decreased in CAM assay after LV-RNAi transfection. Meanwhile, no obvious cell viability and apoptosis changes were found in MTT assay and Flow Cytometry, respectively. LV-RNAi inhibited pancreatic carcinoma angio- genesis and growth by downregulating the expression of VEGF and CD34. These findings demonstrate that Ang2 gene silencing may exert a anti-angiogenesis effect in vitro and in vivo, and Ang2 targeted gene therapy has the potential to serve as a novel way for pancreatic carcinoma treatment.

Upregulation of miR-27a contributes to the malignant transformation of human bronchial epithelial cells induced by SV40 small T antigen.

The introduction of the Simian virus 40 (SV40) early region, the telomerase catalytic subunit (hTERT) and an oncogenic allele of H-Ras directly transforms primary human cells. SV40 small T antigen (ST), which forms a complex with protein phosphatase 2A (PP2A) and inhibits PP2A activity, is believed to have a critical role in the malignant transformation of human cells. Recent evidence has shown that aberrant microRNA (miRNA) expression patterns are correlated with cancer development. Here, we identified miR-27a as a differentially expressed miRNA in SV40 ST-expressing cells. miR-27a is upregulated in SV40 ST-transformed human bronchial epithelial cells (HBERST). Suppression of miR-27a expression in HBERST cells or lung cancer cell lines (NCI-H226 and SK-MES-1) that exhibited high levels of miR-27a expression lead to cell growth arrested in the G(0)-G(1) phase. In addition, suppression of miR-27a in HBERST cells attenuated the capacity of such cells to grow in an anchorage-independent manner. We also found that suppression of the PP2A B56γ expression resulted in upregulation of miR-27a similar to that achieved by the introduction of ST, indicating that dysregulation of miR-27a expression in ST-expressing cells was mediated by the ST-PP2A interaction. Moreover, we discovered that Fbxw7 gene encoding F-box/WD repeat-containing protein 7 was a potential miR-27a target validated by dual-luciferase reporter system analysis. The inverse correlation between miR-27a expression levels and Fbxw7 protein expression was further confirmed in both cell models and human tumor samples. Fbxw7 regulates cell-cycle progression through the ubiquitin-dependent proteolysis of a set of substrates, including c-Myc, c-Jun, cyclin E1 and Notch 1. Thus, promotion of cell growth arising from the suppression of Fbxw7 by miR-27a overexpression might be responsible for the viral oncoprotein ST-induced malignant transformation. These observations demonstrate that miR-27a functions as an oncogene in human tumorigenesis.

α4 is highly expressed in carcinogen-transformed human cells and primary human cancers.

A regulator of the protein phosphatase 2A (PP2A), α4, has been implicated in a variety of functions that regulate many cellular processes. To explore the role of α4 in human cell transformation and tumorigenesis, we show that α4 is highly expressed in human cells transformed by chemical carcinogens including benzo(a)pyrene, aflatoxin B(1), N-methyl-N'-nitro-N-nitrosoguanidine, nickel sulfate and in several hepatic and lung cancer cell lines. In addition, overexpression of α4 was detected in 87.5% (74/80) of primary hepatocellular carcinomas, 84.0% (21/25) of primary lung cancers and 81.8% (9/11) of primary breast cancers, indicating that α4 is ubiquitously highly expressed in human cancer. Functional studies revealed that elevated α4 expression results in an increase in cell proliferation, promotion of cell survival and decreased PP2A-attributable activity. Importantly, ectopic expression of α4 permits non-transformed human embryonic kidney cells (HEKTER) and L02R cells to form tumors in immunodeficient mice. Furthermore, we show that the highly expressed α4 in transformed cells or human tumors is not regulated by DNA hypomethylation. A microRNA, miR-34b, that suppresses the expression of α4 through specific binding to the 3'-untranslated region of α4 is downregulated in transformed or human lung tumors. Taken together, these observations identify that α4 possesses an oncogenic function. Reduction of PP2A activity due to an enhanced α4-PP2A interaction contributes directly to chemical carcinogen-induced tumorigenesis.

Cloning, expression and characterization of the serine protease gene from Chaetomium thermophilum.

AIMS: Microbial proteases play an essential role in scientific research and commercial applications. This study is to clone, sequence, and express a thermostable protease gene from the thermophilic fungi Chaetomium thermophilum and to generate yeast strains expressing C. thermophilum protease suitable for industrial applications. METHODS AND RESULTS: Degenerate primers were designed based on the conserved domain of other identified serine proteases and cDNA fragment of C. thermophilum gene pro was obtained through reverse transcriptase-polymerase chain reaction (RT-PCR). The full-length cDNA of 2007 bp was generated using RACE amplification. The cDNA contains an open reading frame of 1596 bp encoding 532 amino acids. Sequence analysis of the deduced amino acid sequence revealed high homology with the catalytic domains of the subtilisin serine proteases. The C. thermophilum gene pro was expressed in Escherichia coli BL21 (DE3) and Pichia pastoris, respectively and soluble protein was obtained in P. pastoris. The expressed protease was secreted into the culture medium by the yeast in a functional active form and the purified recombinant protease exhibits optimum catalytic activity at pH 8.0 and 60 degrees C. The enzyme is stable at 60 degrees C. The integration of gene pro into P. pastoris genome is stable after 10 generations and the yeast transformants showed a consistent protease expression. CONCLUSIONS: Gene pro encoding a serine protease from C. thermophilum was cloned, sequenced, and overexpressed successfully in P. pastoris. The expressed protease was purified and the properties of the recombinant protease are characterized. SIGNIFICANCE AND IMPACT OF THE STUDY: Chaetomium thermophilum is a soil-borne thermophilic fungus and the protease cloned from it is stable in a high temperature and a wide rage of pH. The overexpression of the enzyme in a mesophilic micro-organism offers a potential value for scientific research and commercial applications.