[Comparison of analgesic effect of modified superior trunk block and traditional interscalene brachial plexus block with liposomal bupivacaine after arthroscopic rotator cuff repair].

Objective: To compare the postoperative analgesic effect of modified superior trunk block and traditional interscalene brachial plexus block in arthroscopic rotator cuff repair. Methods: A total of 40 patients undergoing arthroscopic rotator cuff repair in the Second Affiliated Hospital of Wenzhou Medical University from October to November 2023 were prospectively included, whose American Society of Anesthesiologists (ASA) grade were Ⅰ-Ⅱ. They were divided into modified superior trunk block group (group S) and interscalene brachial plexus block group (group I) by random number table according to different nerve block methods, with 20 cases in each group. Local anesthetics was a mixture of 1.33% liposomal bupivacaine and 0.5% levobupivacaine hydrochloride injection in equal volume. Patients in group S were injected 5 ml mixture for ultrasound-guided modified superior trunk block, and patients in group I were injected with 15 ml mixture for ultrasound-guided traditional interscalene block respectively. Both groups underwent superficial cervical plexus block (5 ml mixture). Standardized general anesthesia and standardized postoperative analgesia were followed. The primary outcome measures included 48 h resting numerical rating scale (NRS) scores after surgery and the incidence of hemidiaphragmatic paralysis (HDP) at 30 min after block. The secondary outcome measures included resting NRS scores during the post anesthesia care unit (PACU), 12, 24, and 36 h after surgery, postoperative opioid consumption and satisfaction with analgesia, pulse oxygen saturation (SpO2) at 30 min after block, sensory and motor block duration, and the incidence of perioperative adverse reactions. The non-inferiority cut-off value of resting NRS scores for patients in group S was set as"1 point"at each observation time point after surgery. Results: In group S, one patient was excluded because the target nerve was blocked by the subclavian vein and could not be blocked, nineteen patients [11 males and 8 females, aged (52.2±9.0) years] were eventually included. In group I, there were 7 males and 13 females, aged (55.0±5.1) years. Resting NRS scores of group S and Group I at 48 h after surgery were 0 (0, 0) and 0 (0, 0.8) point, respectively, with no statistical significance (P>0.05). The median difference was 0 (95%CI:0-0) point and the upper 95%CI was 0 point, which was lower than the preset non-inferiority cut-off value"1 point"(non-inferiority P<0.001). The incidence of HDP in group S and group I were 5% (1/19) and 75% (15/20), respectively, with statistically significant (P<0.001). There were no significant differences in resting NRS scores at PACU and 12, 24, 36 h after surgery, opioid dosage, satisfaction with analgesia, SpO2 at 30 min after block, sensory and motor block duration between two groups (all P>0.05). No respiratory adverse events such as hypoxemia and airway spasm occurred in two groups after extubation. One patient in group I showed symptoms of breath shortness when entering PACU, and 3 patients felt uncomfortable due to prolonged numbness and weakness of the blockade limb (>2 days). No nerve block procedures and opioid drugs relative adverse reactions and no neurological complications happened in both groups. Conclusion: Liposomal bupivacaine usage for modified superior trunk block can provide long-term postoperative analgesic effects which is noninferior to traditional interscalene brachial plexus block and causes less HDP in patients undergoing arthroscopic rotator cuff repair.

Acute pulmonary embolism caused by highly aggregated intravenous immunoglobulin.

BACKGROUND AND OBJECTIVES: Six patients died and one patient survived following infusion of a specific lot of intravenous immunoglobulin (IVIG) within half an hour in May 2008. This study elucidated the underlying pathogenesis. MATERIALS AND METHODS: A variety of protein fractionation and identification approaches were employed to determine the abnormal components in IVIG products obtained from the hospital where the patients were treated. Animal studies using mice and monkeys were conducted to elucidate the pathophysiological mechanisms. In animal experiments, the effect and distribution of immunoglobulin was investigated using HE staining and immunohistochemistry (IHC) separately, while platelets and fibrinogen depletion were utilized to determine a possible link between thromboembolism formation in animals and the lethal effect of the IVIG. The size and distribution of the protein aggregates were determined with Coulter Counter Multisizer-3 after the dilution of the IVIG with plasma, and the lethal effect of the protein aggregates was simulated with artificial microparticles. RESULTS: The IVIG retrieved from the hospital was found to have striking similarities to the heat-treated IVIG in terms of protein aggregation profiles and lethal effects. Post-mortem examination indicated that immunoglobulin aggregates were mainly found in the lung of the animals, while depletion of platelets and fibrinogen from the IVIG preparations failed to prevent the death of the animals. Similar amount of artificial microparticles caused animal death in similar fashion. CONCLUSIONS: Our findings indicate that the retrieved IVIG exerted its lethal effects by blocking the pulmonary circulation without markedly altering the coagulation cascade or immunological events.

Lattice distortion enabling enhanced strength and plasticity in high entropy intermetallic alloy.

Intermetallic alloys have traditionally been characterized by their inherent brittleness due to their lack of sufficient slip systems and absence of strain hardening. However, here we developed a single-phase B2 high-entropy intermetallic alloy that is both strong and plastic. Unlike conventional intermetallics, this high-entropy alloy features a highly distorted crystalline lattice with complex chemical order, leading to multiple slip systems and high flow stress. In addition, the alloy exhibits a dynamic hardening mechanism triggered by dislocation gliding that preserves its strength across a wide range of temperatures. As a result, this high-entropy intermetallic circumvents precipitous thermal softening, with extensive plastic flows even at high homologous temperatures, outperforming a variety of both body-centered cubic and B2 alloys. These findings reveal a promising direction for the development of intermetallic alloys with broad engineering applications.

[Construction of natural population cohort on telephone follow-up management quality control system and discussion regarding critical issues by REDCap system].

With completing a baseline survey of a large natural population cohort, conducting regular follow-up has become a key factor in further improving the quality of cohort construction and ensuring its sustainable development. Typical cohort follow-up methods include repeat surveys, routine monitoring, and community-oriented surveillance. However, in practical applications, there are often issues such as high costs, difficulty, and high error rates. Telephone follow-up is an important supplementary method to the methods mentioned above, as it has the characteristics of low cost, fast response, and high quality. However, the with difficult organization, quality control is challenging, response rates are low, and management levels vary widely, which limits its widespread use in large-scale population cohort studies. Given the above problems, this study draws on customer relationship management based on the actual needs of the China Northwest Cohort follow-up. It relies on the REDCap electronic data collection platform to build a telephone follow-up management and quality control system. Targeted solutions are provided for key issues in telephone follow-up implementation, including organizational structure, project management, data collection, and process quality control, to improve the quality control level of telephone follow-up comprehensively and thereby enhance the quality and efficiency of follow-up. We hope to provide standardized follow-up programs and efficient quality control tools for newly established and existing cohort studies.

Optical and electrical properties of Ga-doped ZnO nanorod arrays fabricated by thermal evaporation.

Well-aligned Ga-doped ZnO nanorod arrays with high optical and electrical property were fabricated by catalyst-free thermal evaporation on p-silicon substrate. As the Ga/Zn atom ratio in the source material was tuned from 0 to 0.2, wurtzite structure ZnO nanorod arrays were realized with length of -6 microm and growth direction along c-axis. With the addition of Ga, the intensity of the near-band-edge emission was enhanced and the deep-level emissions maintained neglectable. As the Ga/Zn atom ratio increased from 0 to 0.1, the red shift of the near-band-edge emission occurred due to Ga-doping induced band gap renormalization effect related with the enhancement of the carrier density, while the blue shifts of the emission were found once the Ga/Zn ratio is higher than 0.1 resulting from Burstein-Moss effect. The configuration of the vertical-aligned Ga-doped ZnO nanorod arrays on p-Si substrate makes it straightforward for the fabrication of p-n nanodiode, which shows an excellent rectifying characteristic with threshold voltage as low as -4.7 V with the Ga/Zn atomic ratio of 0.2.

[A predictive model based on risk factors for early mortality in patients with newly diagnosed multiple myeloma].

Objective: To investigate risk factors for early mortality (EM) in patients with newly diagnosed multiple myeloma (NDMM) and to build an EM-predictive model. Methods: In a cohort of 275 patients with NDMM, risk factors for EM at 6, 12, and 24 months after diagnosis (EM6, EM12, and EM24, respectively) were determined to establish a model to predict EM. Results: The rates of EM6, EM12, and EM24 were 5.5% , 12.7% , and 30.2% , respectively. The most common cause for EM was disease progression/relapse, accounting for 60.0% , 77.1% , and 84.3% of EM6, EM12, and EM24, respectively. EM6 was associated with corrected serum calcium >2.75 mmol/L and platelet count <100×10(9)/L, whereas risk factors for EM12 included age >75 years, ISS Ⅲ, R-ISS Ⅲ, corrected serum calcium >2.75 mmol/L, serum creatinine >177 µmol/L, platelet count <100×10(9)/L, and bone marrow plasma cell ratio ≥ 60% . In addition to the risk factors for EM12, EM24 was also associated with male sex and 1q21 gain. By multivariate analysis, age >75 years, platelet count <100×10(9)/L, and 1q21 gain were independent risk factors for EM24 but there were no independent risk factors significantly associated with EM6 and EM12. Using a scoring system including these three risk factors, a Cox model for EM24 was generated to distinguish patients with low (score<3) and high (score ≥ 3) risk. The sensitivity and specificity of the model were 20.7% and 99.2% , respectively. Further, an internal validation performed in a cohort of 183 patients with NDMM revealed that the probability of EM24 in high-risk patients was 26 times higher than that in low-risk patients. Moreover, this model was also able to predict overall survival. The median overall survival of patients with scores of 0, 1, 2, 3, 4, and 5 were 59, 41, 22, 17.5, and 16 months, respectively. Conclusion: In the study cohort, the EM6, EM12, and EM24 rates were 5.5% , 12.7% , and 30.2% , respectively, and disease progression or relapse were main causes of EM. An EM24-predictive model built on three independent risk factors for EM24 (age>75 years, platelet count<100×10(9)/L, and 1q21 gain) might predict EM risk and overall survival.

MicroRNA-28-5p regulates glioma cell proliferation, invasion and migration by targeting SphK1.

The article "MicroRNA-28-5p regulates glioma cell proliferation, invasion and migration by targeting SphK1, by H.-S. Chen, A.-Q. Lu, P.-Y. Yang, J. Liang, Y. Wei, Y.-W. Shang, Q. Li, published in Eur Rev Med Pharmacol Sci 2019; 23 (15): 6621-6628-DOI: 10.26355/eurrev_201908_18551-PMID: 31378904" has been withdrawn from the authors stating that "after our follow-up experiments and in-depth research, we found that the previous experimental data had some loopholes and deviations. After the experiment was improved, we found that some experimental data could not be repeated again. To avoid academic adverse effects, we ask the magazine to withdraw the manuscript". The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/18551.

Protective effects of gliclazide on high glucose and AGEs-induced damage of glomerular mesangial cells and renal tubular epithelial cells via inhibiting RAGE-p22phox-NF-kB pathway.

OBJECTIVE: Gliclazide is one of the most widely used therapeutic drugs for diabetes. As a second-generation sulfonylurea oral hypoglycemic drug, it can lower blood glucose level and delay the occurrence and development of diabetic nephropathy (DN). However, the underlying mechanism remains unclear. Therefore, the aim of this study was to explore whether gliclazide had protective effects on high glucose and advanced glycation end products (AGEs)-induced injury of human mesangial cells (HMCs) and renal tubular epithelial cells. MATERIALS AND METHODS: HMC and renal tubular epithelial cell lines [human kidney 2 (HK-2)] were cultured in vitro. All cells were then divided into the follow groups: 1) blank control group (5.6 mmol/L glucose), 2) AGEs group [400 µg/mL AGE-bovine serum albumin (AGE-BSA)], 3) high glucose group (25 mmol/L glucose), 4) gliclazide + AGEs group (400 µg/mL AGE-BSA + 20 µmol/L gliclazide) and 5) gliclazide + high glucose group (25 mmol/L glucose + 20 µmol/L gliclazide). Cell counting kit-8 (CCK-8) assay was adopted to determine cell viability. Flow cytometry was used to detect cell apoptosis. The levels of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were measured as well. Furthermore, the mRNA expressions of receptor for AGE (RAGE), p22phox and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) were measured via fluorescence quantitative Real-time polymerase chain reaction (qRT-PCR). RESULTS: Compared with control group, significantly accelerated apoptosis of HMCs and HK-2, increased MDA level, decreased SOD and GSH-Px levels, and up-regulated mRNA expressions of RAGE, p22phox and NF-κB were observed in HMCs and HK-2 of high glucose group and AGEs group. Meanwhile, there were obviously alleviated apoptosis of HMCs and HK-2, decreased MDA level, increased SOD and GSH-Px levels, as well as down-regulated mRNA expressions of RAGE, p22phox and NF-κB in HMCs and HK-2 of gliclazide group compared with high glucose and AGEs group. Furthermore, significant correlations were found between the mRNA expression of RAGE and the apoptosis rate of HMCs and HK-2 (HMCs: r=0.701, p=0.004 and HK-2: r=0.633, p=0.011). CONCLUSIONS: Gliclazide has protective effects on high glucose and AGEs-induced damage of glomerular mesangial cells and renal tubular epithelial cells via inhibiting RAGE-NADPH oxidase-NF-kB pathway.

MicroRNA-28-5p regulates glioma cell proliferation, invasion and migration by targeting SphK1.

OBJECTIVE: MicroRNAs (miRNAs) are a conserved class of endogenous and short non-coding RNAs that post-transcriptionally regulate the expression of genes involved in diverse cellular processes. MiR-28-5p has been reported to be associated with several cancers, including human glioma. However, the roles of miR-28-5p in glioma development are poorly understood. MATERIALS AND METHODS: Sixteen human glioma tissues and paired adjacent normal tissues were acquired through the Gansu Provincial Hospital. We performed quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) to detect the miR-28-5p expression between 16 paired adjacent normal and glioma tissues, as well as the miR-28-5p expression between normal human astrocytes cells and five glioma cell lines. To examine the functional roles of the downregulated miR-28-5p in glioma, cell viability and colony formation assays were performed for the analysis of cell growth. We overexpressed miR-28-5p by transient transfection of miRNAs mimics and performed the transwell Matrigel invasion assay and transwell migration (without Matrigel) assay. To investigate the roles of miR-28-5p in SphK1 expression, Western blot and Real Time-Polymerase Chain Reaction assays were performed. RESULTS: In this work, we demonstrated that miR-28-5p is downregulated in glioma tissues compared to the adjacent normal tissues. Functional studies showed that miR-28-5p overexpression inhibited the cell viability, colony formation and proliferation; meanwhile, it induced the cell apoptosis. The transwell invasion assay indicated that miR-28-5p blocked the invasion and migration of glioma cells. SphK1 (Sphingosine kinase 1 antibody) is predicted as a targeted candidate of miR-28-5p. Then, the Luciferase reporter assay, Western blot and Real Time-Polymerase Chain Reaction (PCR) validated that miR-28-5p negatively regulated SphK1 expression by directly targeting its 3'untranslated regions (3'UTR) in U87 cells. Furthermore, rescue assay suggested that overexpression of SphK1 without its 3'UTR could prevent the miR-28-5p from inducing the inhibition of glioma tumor cells. CONCLUSIONS: Our findings showed that miR-28-5p could suppress the growth, invasion and migration of glioma cells by suppressing the SphK1 expression. The results demonstrated that miR-28-5p might serve as an important potential therapeutic target for glioma.

[Prognostic analysis of 182 newly diagnosed multiple myeloma patients with high risk cytogenetic abnormalities].

Objectives: To evaluate the clinical characteristics and prognosis of high risk cytogenetic abnormalities (HRCA) and various combinations of cytogenetic abnormality in patients with newly-diagnosed multiple myeloma (NDMM) . Methods: This retrospective study collected 182 NDMM patients in the First Affiliated Hospital of Jilin University between Nov. 2009 and May 2018. HRCA included 1q+, del (17p) , t (4;14) , and t (14;16) detected by FISH, and non-HRCA included del (13q) , t (11;14) detected by FISH. The clinical characteristics among three groups, including cases who carrying a single HRCA, 1 HRCA in combination with non-HRCA and cases carrying two or more HRCAs (double/triple-hit) were observed. Kaplan-Meier curve was used to analyze both progression-free survival (PFS) and overall survival (OS) for the three groups. Results: The survivals of patients with 1 HRCA in combination with non-HRCA were similar to those with two or more HRCAs (double/triple-hit) , the median PFS (mPFS) was 19.1 m vs 12.1 m (P=0.248) and median OS (mOS) was 29.6 m vs 29.3 m (P=0.774) . Furthermore, the prognosis of these two groups were both inferior to patients with a single HRCA, respectively. (mPFS: 32.2 m, P=0.040, P=0.001; mOS: 42.3 m, P=0.021, P=0.041) . Strikingly, both the mPFS and the mOS of patients with 1 HRCA in combination with non-HRCA (regardless of high risk or not) were significantly shorter than that of cases with a single HRCA (mPFS: 15.1 m vs 32.2 m, HR=2.126, 95%CI 1.176-3.843, P=0.005; mOS: 29.3 m vs 42.3 m, HR=1.442, 95%CI 0.705-2.950, P=0.011) . Conclusion: It is of prognostic significance value for detecting double/triple-hit based on FISH cytogenetics in NDMM.

[Adverse effects of double-hit combining ISS-â ¢ stage and 1q gain or del (17p) on prognosis of patients with newly diagnosed multiple myeloma].

Objective: To evaluate the prognostic significance of combining ISS-Ⅲ and high risk cytogenetic abnormalities [HRCAs, including 1q gain/amplification and del (17p) ] in patients with newly-diagnosed multiple myeloma (NDMM) . Methods: The clinical characteristics and relevant variables were retrospectively analyzed in a total of 270 NDMM patients diagnosed between November 2009 and May 2018. ISS-Ⅲ stage and HRCAs [detected by FISH, including 1q gain/amplification and del (17p) ] were defined as risk factors (hit) . Based to the number of hit per case, these patients were divided into four groups carrying 0 to 3 risk factors, respectively. Progress-free survival (PFS) and overall survival (OS) were then analyzed using the Kaplan-Meier estimator. Results: Patients who carried single hit (n=120, 44.4%) had shorter median PFS (23.0 vs 28.9 months; P>0.05) and OS (42.3 vs 53.7 months; P>0.05) than those with no risk factors (n=66, 24.4%) . Of note, the outcome of patients who had two or more risk factors (double/triple, n=84, 31.1%) was much worse than those with either no or one risk factor, indicated by significantly reduced median PFS (14.5 months; HR=1.584, 95%CI 1.082-2.319; P=0.003 for double/triple vs single hit) and OS (18.4 months, HR=2.299, 95%CI 1.485-3.560; P<0.001 for double/triple vs single hit) . Strikingly, patients who had three risk factor (triple hit, n=5, 1.9%) displayed the poorest survival with extraordinarily shorter PFS (0.9-15.1 months) and OS (0.9-18.9 months) compared to those carrying two risk factors (double hit) . Analogous results were obtained when different combinations of ISS stages and HRCAs were analyzed. Conclusion: These results suggest a potential but rather important role of combining multiple (e.g. double or triple) adverse factors determined via the routine ISS staging and FISH detection of cytogenetic abnormalities in risk stratification and prognostic prediction, which might be helpful to identify high risk patients more precisely at diagnosis. It also raised a possibility that a small group of ISS-Ⅲ patients carrying both 1q gain/amplification and del (17p) might represent an "extremely-high risk" subset of MM.

[The prognostic significance of dynamic monitoring of minimal residual disease (MRD) status in patients with newly-diagnosed multiple myeloma].

Objective: To evaluate the prognostic value of kinetic changes in minimal residual disease (MRD) status, as well as its relationship with risk stratification, therapeutic response and treatment in patients with newly-diagnosed multiple myeloma (MM) . Methods: A total of 135 patients with newly-diagnosed MM were screened, and 105 patients who achieved VGPR or more as the best responses were included into this study. The MRD status was determined by multiparameter flow cytometry (MFC) at multiple intervals after two cycles of treatment until clinical relapse, death, or last follow-up. The statistical methods included Kaplan-Meier analysis, Cox regression, etc. Results: ①In all 135 patients, 57.8% (78/135) patients achieved MRD negativity (MRD(-)) after treatment. In 105 patients who achieved VGPR and thus included in this study, the MRD(-) rate was 72.4% (76/105) , with a median interval of 3 months from starting treatment to achievement of MRD(-) status. ②The 2-year PFS rate of patients with MRD(-) status was significantly higher than that of MRD(+) status (62.2% vs 41.3%, P=0.001) , while MRD persistence (MRD(+)) was an independent factor for poor prognosis (multivariate analysis for PFS: P=0.044, HR=3.039, 95%CI 1.029-8.974) . ③Loss of MRD(-) status (i.e., MRD reappearance) showed inferior outcomes compared with MRD sustained negative ones, the PFS was 18 months versus not reach (P<0.001) and the OS was not reach for both (P=0.002) . ④The 2-year PFS and OS rates of patients with duration of MRD(-)status≥12 months were significantly higher than those of the control group (PFS: 77.7% vs 36.7%, P<0.001; OS: 96.4% vs 57.9%, P<0.001 respectively) . Duration of MRD(-) status was associated with a marked reduction in risk of relapse or death (univariate analysis for PFS: P<0.001, HR=0.865, 95%CI 0.815-0.918; for OS: P=0.001, HR=0.850, 95%CI 0.741-0.915 respectively) . ⑤Moreover, even in patients carrying high-risk cytogenetic abnormalities (CA) or ineligible for ASCT, MRD negativity remained its prognostic value to predict PFS (high-risk CA medianPFS: not reach vs 19 months, P=0.006; ineligible for ASCT medianPFS: not reach vs 25 months, P=0.052 respectively) . ⑥Last, treatment with the bortezomib-based regimens contributed to prolonged MRD(-) duration (median MRD(-) duratio: 25 months vs 10 months, P=0.034) . Conclusion: Our findings supported MRD(+) status as an independent poor prognostic factor in MM patients, which implicated that duration of MRD(-) status also played a significant role in evaluation of prognosis, while loss of MRD(-)status might serve as an early biomarker for relapse. Therefore, monitoring of MRD kinetics might more precisely predict prognosis, as well as guide treatment decision, especially for when to start retreatment in relapsed patients.

[Effect of 1q21 amplification on bortezomib therapeutic response and prognosis of newly diagnosed multiple myeloma patients].

Objective: To investigate the effect of 1q21 amplification (1q) on the therapeutic response and prognosis of bortezomib(Btz) in the treatment of newly diagnosed multiple myeloma (MM) patients. Methods: A total of 180 newly diagnosed MM were included for analyses of clinical characteristics, cytogenetics, objective response rate (ORR), progression-free survival (PFS) and overall survival (OS), retrospectively. Gene expression profiling (GEP) was analyzed using publicly available R2 platform. Results: ① In 180 patients, 1q was found in 51.1% cases. Of them, 174 patients had complete follow-up data, including 88 cases with 1q and 86 without 1q (non-1q). ②Incidence of 1q was positively associated with percentage of IGH rearrangement (72.2%, P=0.017) and 1p deletion (1p) (27.8%, P=0.040). ③ The median PFS was 15.0 and 20.3 months for the 1q group and non-1q group, and the median OS was 29.4 and 44.0 months, respectively. Both PFS and OS of 1q group was significantly shorter than those of the non-1q group (P=0.029 and 0.038, respectively). Multivariate analysis further revealed that 1q was an independent prognostic factor for both PFS (HR=1.910, 95% CI 1.105-3.303, P=0.020) and OS (HR=2.353, 95% CI 1.090-5.078, P=0.029). ④ In 91 evaluable cases with 1q, very good partial remission (VGPR) rate was higher after treatment with Btz than those without Btz (62.1% vs 40.0%, P=0.032). Of note, the patients with 1q who received auto-HSCT after induction with Btz had significantly longer PFS than those without auto-HSCT (19 months vs 13 months, P=0.048). ⑤GEP analysis revealed that 1q21 amplification predominantly up-regulated expression of >50% genes within 1q21 region, and also altered expression of 28% genes in chromosome 1 and 10% genes in whole genome, particularly related to DNA repair and cell cycle. Conclusions: 1q is an independent adverse prognostic factor in patients with newly diagnosed MM. It is often associated with 1p deletion and IGH rearrangement. Patients with 1q respond well to Btz-based regimen, but they fail to gain long-term benefit from this treatment itself. However, auto-HSCT following Btz induction might improve survival of patients with 1q, suggesting a potential strategy to treat this high-risk subset of MM. GEP analysis warrants further attention in understanding the mechanisms underlying the high-risk of 1q.

Comparative proteome analysis of rat brain and coronary microvascular endothelial cells.

The endothelium of different organs displays a remarkable heterogeneity, although it presents many common functional and morphological features. However, despite our knowledge of heterogeneity among endothelial cells from different sites, the differences between brain microvascular endothelial cells (BMEC) and coronary microvascular endothelial cells (CMEC) are poorly defined. The aim of this study was to investigate whether BMEC are distinct from CMEC at the protein level. Using the proteomic approach, we comparatively analyzed the proteome of cultured BMEC and CMEC. We reproducibly separated over 2000 polypeptides by using two-dimensional electrophoresis (2-DE) at pH range of 3-10. Using PDQuest software to process the 2-DE gel images, forty-seven protein spots were differentially expressed in the two-endothelial cells. Of these, thirty-five proteins are highly expressed in BMEC, whereas twelve proteins are highly expressed in CMEC. Fifteen proteins in BMEC and seven proteins in CMEC were identified with high confidence by matrix-associated laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS). Our data suggested that BMEC and CMEC were different in several aspects including cytokine and growth-related molecules, stress-related proteins, metabolic enzymes, signal transduction proteins and others. The identification of a set of proteins preferentially expressed in BMEC and CMEC provided new data on the heterogeneity of the endothelium.

Integrated advanced natural wastewater treatment system for small communities.

A study was conducted to evaluate the nutrient removal capability of an existing and successfully operated overland flow and wetland wastewater treatment system following a waste stabilization pond. Seasonal temperature effects on performance were also investigated. The treatment system studied consists of a two-cell waste stabilization pond followed by an overland flow system and a wetland system. The influent and effluent samples were analyzed for BOD5, suspended solids (SS), pH, temperature, ammonia nitrogen, nitrate nitrogen, and total phosphorus. The results of the study indicate that the combined pond, overland flow and wetland system provided excellent treatment of municipal wastewater. The overall average BOD5 removal by the entire treatment system was about 90.0% and the overall average suspended solids removal was about 93.4%. The ammonia nitrogen and total phosphorus removal efficiencies of the entire treatment system were 90.7% and 84.2%, respectively.

Development of an effective sample preparation approach for proteomic analysis of silkworm eggs using two-dimensional gel electrophoresis and mass spectrometry.

Sample preparation is still the first and important step toward successful two-dimensional gel electrophoresis (2DE) and identification in proteomics study. The 2DE profiling of eggs of silkworm species by using conventional one-step extraction, however, is unsatisfactory because high-abundance proteins such as egg-specific protein (ESP) and No 30 family (30 KP) in the extract lead to difficulties in detecting most of biologically relevant proteins. Based on the tendency of these abundant proteins to be soluble in Tris-HCl buffer, we report herein a robust approach in which the extract enriched in ESP and 30 KP was fractionationed and mixed with the re-extract of residual pellet in an optimal proportion. In comparison with the one-step method, the 2DE pattern was improved by this new method with over one-third enhancement in spots. A total of 48 unique proteins obtained have been furthermore identified by mass spectrometry (MS) and MS/MS. The identified proteins are found to include heat shock proteins families, ribosomal proteins, disulfide isomerase proteins, Glutathione S-transferase, and elongation factor, etc., which are mainly involved in some important processes. To our knowledge, this is the first time that the several proteins have been detected in silkworm eggs by proteomics means. This simple and reproducible approach would raise the opportunity of discovering and identifying more biomarkers and determining their possible roles in further studies.

Removal of organic and nitrogen and molecular weight distribution of residual soluble organic from entrapped mixed microbial cells and activated sludge processes.

The simultaneous removal of organic and nitrogen and molecular weight distribution (MWD) of residual soluble chemical oxygen demand (RSCOD) in final effluent were investigated using entrapped mixed microbial cells (EMMC) and conventional activated sludge processes. Two different types of processes using EMMC carriers demonstrated better organic and nitrogen removal performance because of the high solids retention time (SRT) compared with the activated sludge process. Regarding the RSCOD, the longer SRT process (EMMC) was affected by reducing the hydraulic retention time, resulting in the increase of high-molecular-weight materials. On the other hand, reducing the aeration period had significantly affected the MWD in the shorter SRT process (activated sludge), resulting in an increase of low-molecular-weight materials.

Bio-anaerobic treatability study for PCBs-contaminated oil.

This study investigated the bio-treatability of PCB contaminated oil for the development of design and operational parameters for the bioreactor. Input of external carbon and nutrient source in the aqueous phase was found to be required for the treatment of polychlorinated biphenyls (PCBs)-contaminated oil. Addition of surfactant was investigated for the emulsification of oil to reduce interference of contact with microorganisms and PCBs. The ratio of surfactant to oil was empirically optimized to 1 : 1. The higher PCB removal efficiency was obtained at 30 days of hydraulic retention time (HRT) in the semi-batch reactor study without cell recycle. The removal efficiency measured in mixed liquor was maintained at over 85% on average at 32 +/- 2 degrees C and 30% at 22 +/- 2 degrees C. More than 0.2 g/l/d of the organic loading rate was suggested to be maintained for various PCB loading rates (0.02-0.6 mg-PCB/l/d). For high biomass retaining and easy collection of treated oil, an Anaerobic Sequencing Batch Reactor (ASBR) was investigated. The removal of Aroclor was observed as more than 50% in the oil phase with 3 days reaction time and about 40% in overall phases, i.e. oil, liquid, biomass phases at 22 +/- 2 degrees C. US EPA verification results on the process performance are included in this presentation.

Cloning of a human insulin-stimulated protein kinase (ISPK-1) gene and analysis of coding regions and mRNA levels of the ISPK-1 and the protein phosphatase-1 genes in muscle from NIDDM patients.

Complementary DNA encoding three catalytic subunits of protein phosphatase 1 (PP1 alpha, PP1 beta, and PP1 gamma) and the insulin-stimulated protein kinase 1 (ISPK-1) was analyzed for variations in the coding regions related to insulin-resistant glycogen synthesis in skeletal muscle of 30 patients with non-insulin-dependent diabetes mellitus (NIDDM). The human ISPK-1 cDNA was cloned from T-cell leukemia and placental cDNA libraries and mapped to the short arm of the human X chromosome. Single-strand conformation polymorphism (SSCP) analysis identified a total of six variations in the coding regions of the PP1 genes: two in PP1 alpha at codons 90 and 255; one in PP1 beta at codon 67; and three in PP1 gamma at codons 11,269, and 273, respectively. All were, however, silent single nucleotide substitutions. SSCP analysis of the ISPK-1 gene identified one silent polymorphism at codon 266 and one amino acid variant at codon 38 (Ile-->Ser). This variant was primarily found in one male NIDDM patient. This subject, however, did not exhibit an impairment of muscle insulin-stimulated glycogen synthase activation. No significant differences were found in mRNA levels in muscle of the four genes between 15 NIDDM patients and 14 healthy subjects. Our findings suggest that 1) genetic abnormalities in the coding regions of PP1 alpha, PP1 beta, PP1 gamma, and ISPK-1 are unlikely to be frequently occurring causes of the reduced insulin-stimulated activation of the glycogen synthesis in muscle from the analyzed group of NIDDM patients; 2) the mRNA levels of PP1 alpha, PP1 beta, PP1 gamma, and ISPK-1 are normal in muscle from the NIDDM patients; and 3) putative inherited defects in insulin-stimulated activation of muscle glycogen synthesis in patients with insulin-resistant NIDDM may be located further upstream of ISPK-1 in the insulin action cascade.