[Evaluation of the efficacy of medial open wedge high tibial osteotomy combined with anterior cruciate ligament reconstruction in the treatment of varus knee osteoarthritis with anterior cruciate ligament injury].

Objective: To evaluate the efficacy of medial open wedge high tibial osteotomy (MOWHTO) combined with anterior cruciate ligament (ACL) reconstruction in the treatment of varus knee osteoarthritis (OA) with ACL injury. Methods: A follow-up study. The study retrospectively analyzed the patients underwent MOWHTO combined with ACL reconstruction for treatment of varus knee OA with ACL injury in Tianjin Hospital between April 2018 and September 2022. The preoperative and postoperative posterior slope angle (PSA), hip-knee-ankle angle (HKA), visual analog scale (VAS) pain scores, Lysholm score, International Knee Documentation Committee (IKDC) score, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score, and Tegner score were compared. The follow-up indicators were recorded at 6 weeks, 3 months and 1 year after operation, and the complications were recorded. Results: The study included 32 patients (23 males, 9 females) with a mean age of (50.7±8.4) years. The mean follow-up time was (21.2±4.8) months. PSA increased from 9.2°±1.8° preoperatively to 11.1°±2.4° postoperatively, and HKA increased from 168.7°±2.2° to 181.5°±2.2° (both P<0.01). The indicators such as VAS score (6.8±1.1 vs 1.8±0.4), Lysholm score (52.6±7.1 vs 82.0±6.4), IKDC score (64.7±6.2 vs 80.3±10.0), WOMAC score (51.8±6.3 vs 81.8±6.5), and Tegner score (1.9±0.6 vs 5.0±1.0) were all improved after the operation (all P<0.01). Complications occurred in 5 patients (15.6%), including hematomas, sensory abnormalities, intermuscular vein thrombosis and correction angle loss. Conclusion: MOWHTO combined with ACL reconstruction is a safe and effective approach for the treatment of varus knee OA with ACL injury.

[Research on the protective effect of hearing protective device for workers exposed to noise in a motor manufacturing enterprise].

Objective: To understand the current situation of noise hazard in a motor manufacturing enterprise, and to explore the protective effect of workers wearing hearing protective device and its possible influencing factors. Methods: In November 2021, a total of 179 noise workers wearing hearing protective devices in a motor manufacturing company in a city were selected as research objects. Personal attenuation rating (PAR) of workers wearing hearing protective devices was measured. Baseline PAR was analyzed for different subgroups of basic demographic information, noise exposure, and the use of hearing protective devices to evaluate the effect of the intervention. Baseline PAR was compared using nonparametric tests. Results: There were 179 workers from 35 positions in 4 types of work, and the over-standard noise rate was 51.2% (42/82), among which the noise exposure intensity of motor equipment debugging workers was the highest [94.4 dB (A) ]. Compared the baseline PAR of different characteristics, it was found that the baseline PAR of male workers, workers whose daily noise exposure time were <8 h, workers who had used the hearing protective devices for 10 to 14 years, and workers who thought the hearing protective devices were comfortable were all higher, and the differences were statistically significant (P<0.05). Baseline PAR passing rate was 43.0% (77/179), and PAR of 102 workers who did not pass baseline test increased from 0 (0, 3) dB before intervention to 14 (12, 16) dB after intervention, with statistical significance (P<0.05) . Conclusion: The noise hazard in this motor manufacturing enterprise is serious, and the protective effect of workers wearing hearing protective devices is not good. Gender, daily noise exposure time, years and comfort of wearing hearing protective device are the possible influencing factors of poor protective effect.

[Establishment and validation of a predictive clinical model for postoperative surgical site infection in patients with colorectal surgery].

Objective: To investigate the risk factors of surgical site infection (SSI) after colorectal surgery, and to establish and validate a risk prediction model nomogram. Methods: An observational study was conducted to retrospectively collect data of 6527 patients aged ≥16 years who underwent colorectal surgery in 56 domestic hospitals from March 1, 2021 to February 28, 2022 from the national Surgical Site Infection Surveillance network. The incidence of SSI after surgery was 2.3% (149/6527). According to the ratio of 7:3, 6527 patients were randomly divided into the modeling cohort (4568 cases) and the validation cohort (1959 cases), and there was no statistically significant difference between the two datasets (P>0.05). Univariate analysis was performed using t test /Mann-Whitney U test /χ2 test. Multivariate analysis was performed using binary logistic regression to establish a preliminary model and select variables using Lasso analysis to establish an optimized model nomogram. The discrimination and calibration of the model were evaluated by ROC curve, calibration curve, and Hosmer-Lemeshow test. AUC value>0.7 is considered a good discrimination of the model. The Bootstrap method (repeated self-sampling 1000 times) was used to verify the constructed model internally and externally to evaluate the accuracy of the constructed model. Results: Multivariate analysis showed that history of chronic liver disease (OR=3.626, 95%CI: 1.297-10.137, P<0.001) and kidney disease (OR=1.567,95%CI:1.042-2.357,P=0.038), surgical antibiotic prophylaxis (OR=1.564, 95%CI:1.038-2.357,P=0.035), and emergency surgery (OR=1.432,95%CI: 1.089-1.885, P=0.021), open surgery (OR=1.418, 95%CI:1.045-1.924, P=0.042), preoperative stoma (OR=3.310, 95%CI:1.542-7.105,P<0.001), postoperative stoma (OR=2.323,95%CI: 1.537-8.134,P<0.001), surgical incision type above grade II (OR=1.619,95%CI:1.097-2.375,P=0.014), and each unit increase in total bilirubin (OR=1.003,95%CI:-0.994-1.012, P=0.238), alanine aminotransferase (OR=1.006, 95%CI:1.001-1.011,P=0.032), blood urea nitrogen (OR=1.003,95%CI:0.995-1.011,P=0.310), blood glucose (OR=1.024, 95%CI:1.005-1.043,P=0.027), C-reactive protein (OR=1.007, 95%CI:1.003-1.011,P<0.001), length of incision (OR=1.042, 95%CI:1.002-1.087,P=0.031), surgical duration (OR=1.003,95%CI:1.001-1.005,P=0.017), and surgical blood loss (OR=1.001,95%CI: 1.000-1.002,P=0.045) were risk factors for SSI after colorectal surgery. Each unit increase in albumin level (OR=0.969,95%CI:0.941-0.998,P=0.036) was an independent protective factor for SSI after colorectal surgery. The area under the curve of the optimized model obtained by internal and external validation were 0.768 (95%CI: 0.723-0.813) and 0.753 (95%CI: 0.680-0.832), respectively. The predicted value of the calibration curve was basically consistent with the actual value. Conclusions: The risk prediction model for SSI after colorectal surgery constructed in this study has good discrimination and calibration. The nomogram created in this model can provide an evaluation basis for the observed rate and expected event rate of SSI after clinical colorectal surgery.

[Relationships between lead-induced learning and memory impairments and gut microbiota disturbance in mice].

Objective: To explore the effect of lead exposure on the neurobehavior and gut microbiota community structure in mice. Methods: In August 2019, 64 C57BL/6 mice were randomly divided into 4 groups: control group (0 ppm) , low lead exposure group (20 mg/l) , medium lead exposure group (100 mg/l) and high lead exposure group (500 mg/l) . During the experiment, they were free to eat and drink. The drinking water of the lead exposure group was mixed with lead acetate, and sodium acetate was added in the control group. After 10 weeks of exposure, the Morris water maze was used to test the learning and memory ability of each group of mice, and then they were sacrificed for sampling. ICP-MS was used to detect lead content in whole blood and brain tissue. ELISA was used to determine the level of IL-1ß in mouse serum. 16S rRNA sequencing was used to detect the structural diversity of the intestinal flora in feces, and then the correlation between the flora and behavior indicators was analyzed. Results: In the Morris water maze experiment, compared with the control group, there was no significant difference in the body weight and swimming speed of the mice in the lead exposure groups. The escape latency of the mice in the 100 mg/l and 500 mg/l dose groups was prolonged, and the number of platform crossings decreased (P<0.05) ; meanwhile, the staying time of the mice in the 500 mg/l Pb-treated group in the target quadrant was lower than that of the control group, and the difference was statistically significant (P<0.05) . Compared with the control group, the blood lead content of the mice in each lead exposure group was significantly increased, and the brain lead content of mice in the 500 mg/l dose group was significantly elevated (P<0.05) . The serum IL-1ß levels of mice in each lead exposure group were higher than those of the control group (P<0.05) . At the phylum level, the relative abundance of the Proteobacteria phylum in all of Pb-treated groups was significantly increased (P<0.05) ; at the genus level, Allobaculum, Desulfovibrio, Lachnospiraceae_NK4A136_group, Turicibacter and Ureaplasma were significantly increased (P<0.05) . Among them. The relative abundance of Desuffaoibrio, Turici bacter, and Ureaplasma was negatively correlated with the residence time of mice in the quadrant of the platform (r=-0.32, -0.29, -0.44, P<0.05) . Conclusion: Lead exposure induced learning and memory impairments in mice, which may be related to the disturbance of the gut microbiota.

[Effect and mechanism of TNF-α and etanercept on the invasion ability of extravillous trophoblast cell in URSA patients].

Objective: To investigate the effect and mechanism of tumor necrosis factor α (TNF-α) and its inhibitor etanercept (ETA) on the invasion ability of extravillous trophoblast in patients with unexplained recurrent spontaneous abortion (URSA). Methods: (1) Patients were collected from March to June in 2019. They were divided into the URSA group (n=15) and the normal control group (n=15), according to whether diagnosed with URSA or not. The mRNA expression levels of TNF-α in villi tissue of patients in the two groups were detected by quantitative real-time PCR (qRT-PCR). (2) The mRNA and protein expression levels of matrix metalloproteinase-2 (MMP-2), Slug and CXC chemokine rceptor 4 (CXCR4) in HTR-8/SVneo cells were detected by qRT-PCR or western blot after being stimulated by exogenous TNF-α (0.2, 2, 20 ng/ml) alone or TNF-α along with ETA, or phosphate buffered saline (PBS) as control. (3) The invasion ability of HTR-8/SVneo cells was investigated by transwell test after stimulating by TNF-α alone or TNF-α along with ETA. (4) The mRNA and protein expression levels of MMP-2, Slug and CXCR4 in HTR-8/SVneo cells, which were stimulated by TNF-α (2 ng/ml) alone after nuclear factor-κB (NF-κB) inhibitor, BAY 11-7028, preconditioning, were detected by qRT-PCR or western blot. Results: (1) The mRNA expression level of TNF-α in villi tissue of URSA group (4.10±0.49) was 4.1 times as much as the normal control group (t=10.51, P<0.05). (2) The mRNA and protein expression levels of MMP-2, Slug and CXCR4 in HTR-8/SVneo cells of TNF-α group were significantly lower than those in PBS control group (P<0.05) and those in TNF-α along with ETA group (P<0.05). (3) The invasion ability of HTR-8/SVneo cells in TNF-α group was significantly decreased than PBS group and TNF-α along with ETA group (78±14 vs 373±26 vs 227±44, P<0.05). (4) The mRNA and protein expression levels of MMP-2, Slug and CXCR4 in HTR-8/SVneo cells with BAY 11-7028 preconditioning (mRNA: 1.03±0.10, 1.03±0.06, 1.09±0.08; protein: 1.09±0.03, 1.49±0.03, 1.12±0.03) were significantly higher than without preconditioning after being stimulated by TNF-α (all P<0.05). Conclusions: The expression of TNF-α in the villi of URSA patients is much higher than normal early pregnant women. TNF-α could decrease the capacity of invasion by suppressing the expression of MMP-2, Slug and CXCR4 through NF-κB signaling pathway in extravillous trophoblast cells. While ETA could improve the invasiveness capability of extravillous trophoblast cells through inhibiting the negative effect of TNF-α.

[Significance of circulating tumor cell monitoring in targeted therapy for gastrointestinal stromal tumors].

Objective: To explore the significance of circulating tumor cell (CTC) monitoring in evaluating the efficacy of targeted therapy for gastrointestinal stromal tumor (GIST). Methods: A prospective cohort study was performed. The data of patients with locally advanced GIST or liver metastasis who were admitted to The Affiliated Hospital of Nantong University from August 2013 to December 2018 were collected. Inclusion criteria: (1) patients aged older than 18 years; (2) patients who were diagnosed with GIST based on pathology; (3) patients without surgery, whose preoperative imaging evaluation of GIST found the violations of the surrounding organs or partial transfer of an estimated difficulty to achieve R0 resection, or the maximum diameter of the tumor > 10 cm, or the liver metastasis, or the expectation of higher risk of surgical complications; (4) patients who were treated with the imatinib 400 mg/d for the first time; (5) Eastern Cooperative Oncology Group (ECOG) score of 0-2. Exclusion criteria: (1) genetic testing revealed a D842V mutation in exon 18 of the PDGFRA gene; (2) alanine aminotransferase and/or aspartate aminotransferase > 2.5 times the normal upper limit; (3) serum total bilirubin >1.5 times of normal upper limit; (4) neutrophil count < 1.5×10(9)/L, or platelet count < 75×10(9)/L, or hemoglobin < 60 g/L; (5) creatinine > normal upper limit; (6) patients had serious cardiovascular and cerebrovascular diseases within 12 months before enrollment; (7) female patients were pregnant or lactating; (8) patients suffered from other serious acute and chronic physical or mental problems, and were not suitable for participating in this study judged by researchers. The patients who could not tolerate treatment regimen, or developed serious adverse reactions and did not follow the medication scheme after enrollment were excluded. Before imatinib treatment and 1-month and 2-month after treatment, quantitative PCR was used to detect the DOG-1 expression of monocytes in peripheral blood, and the ratio of DOG-1/ß-actin > 3×10(-5) was used as the CTC positive threshold of GIST. The positive rate of CTC, the efficacy of imatinib treatment (complete response, partial response, stable disease, progressive disease, and occurrence of adverse reactions), and the relationship between CTC positive rate and clinicopathological characteristics of patients were analyzed. Furthermore, the ratio of DOG-1 decrease/baseline DOG-1 after 1-month of treatment was used as an indicator to evaluate whether targeted therapy was effective. The receiver operating characteristic (ROC) curve was rendered, and the area under the curve (AUC) was calculated. Results: A total of 68 GIST patients were enrolled in this study, including 39 cases of locally advanced GIST and 29 cases with liver metastases, 32 males and 36 females with the mean age of (51.2±11.8) (range 31 to 74) years. After 2-month of imatinib treatment, 43 cases were evaluated as partial response, 11 cases as stable disease, and 14 cases as progressive disease, with an effective rate of 79.4% (54/68). During the treatment of imatinib, the incidence of grade 3 or higher adverse reactions was 22.1% (15/68), including 12 cases of grade 3 neutropenia and 3 of grade 4 drug eruption, which were all relieved after conservative treatment. The positive rates of CTC in 68 patients before treatment, 1-month and 2-month after treatment were 66.2% (45/68), 41.2% (28/68) and 23.5% (16/68), respectively. The positive rate of CTC was associated with tumor size, liver metastasis, mitotic count and risk level (all P<0.05). By analyzing the effective group and the ineffective group of targeted therapy, it was found that the positive rate of CTC in the effective group showed a decreasing trend, while the positive rate of CTC in the ineffective group showed an increasing trend. The AUC of predicting the efficacy of targeted therapy for GIST was 0.823 by detecting the change trend of CTC 1-month after treatment (P<0.001). When the DOG-1 content decreased by more than 57.5% 1-month after treatment, it can be used as an indicator to judge the effectiveness of the treatment, whose sensitivity was 72.2% and specificity was 100%. Conclusion: The detection of peripheral blood CTC can evaluate the efficacy of targeted therapy in GIST patients and can provide decision-making basis for further clinical treatment.

Current views of diabetic peripheral neuropathic pain comorbid depression - a review.

Diabetic peripheral neuropathic pain (DPNP) is defined as a pain directly caused by the abnormal somatosensory system in diabetics according to the International Association for the Study of Pain (IASP). The pain has a great impact on the quality of life of diabetic patients. It results in a decline of patients' personal ability to live, which may even cause depression. Over time, the decline in both physical and psychosocial function caused by neuropathic pain may lead to further aggravation of depressive symptoms. This article mainly reviews the prevalence rate, medical expenditure, clinical characteristics, neurobiological features and the treatment of DPNP comorbidity depression, hoping to find the research directions for further study in the future.

[Analysis of complications after medial opening wedge high tibial osteotomy].

Objective: To analyze the incidence of recent complications in patients with osteoarthritis of the knee (OA) after medial opening wedge high tibial osteotomy(MOWHTO) and its influence on clinical effect. Methods: The clinical data of 131 patients with knee OA who received MOWHTO at Department of Sports Medicine and Arthroscopy,Tianjin Hospital from April 2017 to September 2018 were analyzed retrospectively. There were 75 males and 56 females, aged (62.8±5.1) years (range:48 to 70 years). Complications and clinical outcomes of patients were recorded and the proximal medial angle of tibia (MPTA), the International Knee Documentation Committee Subjective Knee Form (IKDC), the Western Ontario and McMaster Universities(WOMAC) Osteoarthritis Index and Knee Injury and Osteoarthritis Outcome score(KOOS) were collected before and 1 year after operation and compared between complication group and non-complication group. Data were analyzed by paired-samples t test, independent samples t test and χ(2) test. Results: The follow-up time was (18.5±3.4) months (range:13 to 22 months). Complications occurred in 22 patients(16.8%), including 8 cases(6.1%) of hematoma, 5 cases(3.8%) of neurosensory abnormality, 4 cases(3.1%) of intramuscular venous thrombosis, 2 cases(1.5%) of deep venous thrombosis, 3 cases(2.3%) of loss of correction angle, 3 cases(2.3%) of superficial infection, 2 cases(1.5%) of deep infection, 2 cases(1.5%) of delayed union of fracture, 1 case(0.8%) of postoperative stiffness, 1 case (0.8%) of hinge point cortex fracture. There were no significant difference in MPTA ((86.5±2.0)° vs. (86.7±2.1)°, t=-0.41, P=0.68) , IKDC ((86.4±4.8) vs.(85.5±6.9), t=0.74, P=0.50) , WOMAC ((87.7±6.5) vs. (86.1±5.8), t=1.16, P=0.25). There were no significant difference in knee scores except for the KOOS pain score ((79.4±4.4) vs. (87.2±5.9), t=-5.90, P<0.01) and sports and recreation score ((83.2±3.0) vs. (88.0±4.7), t=-6.14, P<0.01) . Conclusion: Short-term complications of MOWHTO can be managed appropriately through early diagnosis and individualized treatment and have no significant negative effect on knee function recovery of patients.

MicroRNA-199a-5p regulates glioma progression via targeting MARCH8.

OBJECTIVE: MicroRNA-199a-5p (miR-199a-5p) was reported to play crucial roles in cancer progression. However, its role in glioma remains largely unknown. MATERIALS AND METHODS: RT-qPCR was employed to analyze miR-199a-5p expression level in glioma cell lines. The effects of miR-199a-5p on cell proliferation and invasion were investigated in vitro. RESULTS: We showed that miR-199a-5p expression level was significantly downregulated in glioma cell lines compared with a normal cell. In addition, miR-199a-5p overexpression suppresses glioma cell proliferation and invasion in vitro. Bioinformatic analysis and Luciferase assay suggested that the membrane-associated ring-CH-type finger 8 (MARCH8) was a target of miR-199a-5p. Furthermore, MARCH8 overexpression could partially reverse the effects of miR-199a-5p on glioma cells. CONCLUSIONS: These findings suggested that miR-199a-5p may function as a tumor suppressor in glioma by targeting MARCH8.

[Application of uterine lower part breakwater-like suture operation in placenta previa].

Objective: To explore the efficacy and safety of uterine lower posterior wall breakwater-like suture technique in controlling the intraoperative bleeding of placenta previa. Methods: From June 2016 to June 2017, 47 patients were diagnosed placenta previa in Union Hospital, Tongji Medical College of Huazhong University of Science and Technology. Posterior wall breakwater-like suture technique was used preferentially, as for cases with poor myometrium layer, lower anterior wall stitch suture was used at the same time. Bilateral descending branches of uterine artery ligation and Cook balloon compression of uterine lower segment was conducted when necessary. The clinic data of the 47 cases were analyzed. Results: Thirty cases (63.8, 30/47) were diagnosed placenta inccreta or percreta by ultrasound or MRI preoperatively. Senventeen cases were diagnosed as placenta accreta (36.2%, 17/47) . Thirty-four cases had the previous history of cesarean section. The average cervical canal length of 47 patients was (2.8±0.9) cm. There were 19 cases (40.4%,19/47) with 1 time posterior wall breakwater-like sutured and 16 cases (34.0%,16/47) with 2 or 3 times posterior wall breakwater-like sutured; 12 cases (25.5%,12/47) were treated with anterior wall stitch suture simultaneously.Ten cases (21.3%, 10/47) underwent uterine artery ligation, 17 cases (36.2%, 17/47) underwent COOK balloon compression on the staxis surface of lower segment. None of them had postpartum hemorrhage or performed internal iliac artery embolization. The median blood loss in the operation was 700 ml, the percentiles 25 was 500 ml, and the percentiles 75 was 1 200 ml. The blood loss ≥1 000 ml in 18 (38.3%, 18/47) patients,and the most serious one was 2 500 ml. The median blood transfusion volume (including allogenetic transfusion and autotransfusion) was 450 ml, the percentiles 25 was 228 ml, and the percentiles 75 was 675 ml. The average vaginal bleeding volume was (150±63) ml first day after operation. The mean hospitalization time was (4.7±1.0) days. The mean gestational weeks of pregnancy termination was (36.1±1.5) weeks, and the mean birth weight of newborns was (2 817±492) g. Apgar score:1-minute 7.8±1.1, 5-minute 8.9±0.8. No neonatal death, 16 cases were transferred to neonatal ICU (34.0%, 16/47) mainly for premature delivery and low birth weight. No complication was found in 6 months post-operation. Conclusions: Uterine posterior wall breakwater-like suture technique is a simple, safe and effective way in controlling intraoperative bleeding of placental previa.Lower anterior wall stitch suture could effectively stop bleeding and restore the normal uterine shape. Combined application of various methods could significantly reduce the incidence of postpartum hemorrhage and hysterectomy, and improve maternal and fetal prognosis.

[Clinical value of health-related quality of life evaluation in community patients with hepatitis B].

Chronic hepatitis B (CHB) is a major infectious disease which threatens people's health around the world. Health-related quality of life (HRQoL) has become an important index for evaluating the treatment and prognosis of patients with CHB. Many studies abroad have shown that the HRQoL of patients with CHB is influenced by various factors. Disease severity, clinical symptoms, and low self-efficacy are important influencing factors for patients' quality of life. Anti-viral therapy, proper physical therapy, or psychological intervention can significantly improve the HRQoL of community CHB patients, and HRQoL is even significantly improved in patients undergoing liver transplantation one year ago. These studies provide valid evidence for antiviral therapy for hepatitis B patients and liver transplantation techniques from the psychological level. At present, there still lacks studies on HRQoL of CHB patients in China. This article summarizes the measurement tools for HRQoL widely used in recent years in China and foreign countries, reviews the research advances in HRQoL in community patients with CHB, and discusses the perspectives for future studies.

Role of CASP-10 gene polymorphisms in cancer susceptibility: a HuGE review and meta-analysis.

We investigated a possible association between CASP-10 gene polymorphisms and susceptibility to cancer through a meta-analysis. Eight studies with a total of 29,936 cancer cases and 34,041 healthy controls were included. Meta-analysis results showed that the rs13006529 T carrier was significantly associated with increased cancer risk (OR = 1.17, 95%CI = 1.01-1.36, P = 0.03). However, rs3900115 and rs13010627 showed no association with cancer susceptibility (all P > 0.05). In the subgroup analysis by cancer type, we found that the rs13006529 T carrier was a risk factor for breast cancer (OR = 1.17, 95%CI = 1.01-1.36, P = 0.03). Similarly, no association was found between CASP-10 polymorphisms and susceptibility to lymphoma, myeloma, melanoma, or lung cancer (all P > 0.05). This meta-analysis suggests that the rs13006529 T carrier in the CASP-10 gene might be a risk factor for cancer susceptibility, especially for breast cancer.

Development of the heat treatment system for the 40 T hybrid magnet superconducting outsert.

The heat treatment of Nb(3)Sn coil with the glass fabric insulation is one of the key and critical processes for the outsert solenoids of the 40 T hybrid magnet, which could be wound with cable-in-conduit conductors using the insulation-wind-and-react technique. The manufacturing of the large vertical type vacuum/Ar atmosphere-protection heat treatment system has been completed and recently installed in the High Magnetic Filed Laboratory, Chinese Academy of Sciences. The heat treatment system composed mainly the furnace, the purging gas supply system, the control system, the gas impurities monitoring system, and so on. At present, the regulation and testing of the heat treatment system has been successfully finished, and all of technical parameters meet or exceed specifications.

Inhibition of quorum sensing in Serratia marcescens H30 by molecular regulation.

Quorum sensing in Serratia marcescens, which uses two types of signaling molecules-N-acyl homoserine lactones and furanosyl borate diester-play important regulatory roles in the synthesis of 2,3-butanediol and prodigiosin. In the hope of understanding the effect of quorum sensing on physiologic metabolism, we established two molecular strategies, one to express acyl-homoserine lactone hydrolase to inactivate AI-1 signaling molecule using an expression vector with lactose as the inducer and the other to mutate luxS gene with a suicide plasmid pUTKm2 to inhibit the synthesis of AI-2 signaling molecule.

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.

E2F1 and E2F2 determine thresholds for antigen-induced T-cell proliferation and suppress tumorigenesis.

E2F activity is critical for the control of the G(1) to S phase transition. We show that the combined loss of E2F1 and E2F2 results in profound effects on hematopoietic cell proliferation and differentiation, as well as increased tumorigenesis and decreased lymphocyte tolerance. The loss of E2F1 and E2F2 impedes B-cell differentiation, and hematopoietic progenitor cells in the bone marrow of mice lacking E2F1 and E2F2 exhibit increased cell cycling. Importantly, we show that E2F1 and E2F2 double-knockout T cells exhibit more rapid entry into S phase following antigenic stimulation. Furthermore, T cells lacking E2F1 and E2F2 proliferate much more extensively in response to subthreshold antigenic stimulation. Consistent with these observations, E2F1/E2F2 mutant mice are highly predisposed to the development of tumors, and some mice exhibit signs of autoimmunity.

Apoptosis induction in HCT116 cells by cytochalasins isolated from the fungus Daldinia vernicosa.

We examined the induction of apoptosis by cytochalasin (cc) derivatives (1-14) isolated from the Japanese fungus Daldinia vernicosa to HCT116 human colon cancer cell line based on their cytotoxicity, DNA ladder and DNA fragmentation ratio in agarose gel electrophoresis, and morphological changes. Most cc derivatives tested here induced apoptosis. Particularly cytochalasin 1 (cc1), monoacetate of 1 (cc1Ac), and cc14 were the most potent apoptosis inducers. These apoptotic activities were stronger than that of cytochalasin D as a known apoptosis inducer in HCT116 cell. However, cc4 and cc12 induced necrosis. The structure-activity relationship including their cytotoxicity will be discussed.

Elucidation of strict structural requirements of brefeldin A as an inducer of differentiation and apoptosis.

Brefeldin A (BFA) can induce a wide variety of human cancer cells to differentiation and apoptosis and is in development as an anticancer agent. To elucidate structural requirements for cytotoxicity and induction of differentiation and apoptosis, BFA was structurally modified into various derivatives including 4-epi-BFA in this study. Their inducing activities of apoptosis were evaluated with their cytotoxicities, DNA fragmentation and morphological changes in human colon cancer cell HCT 116. The cytotoxicity of 4-epi-BFA (TX-1923) (IC50 = 60 microM) was 300 times lower than that of BFA (IC50 = 0.2 microM). Furthermore, 4-epi-BFA induced DNA fragmentation and apoptotic morphological changes at much higher concentrations (70 and 50 microM, respectively) than BFA (0.11 and 0.36 microM, respectively). These results indicated that the configuration of 4-hydroxyl group of brefeldin A plays a key role in the cytotoxicity and induction of apoptosis. On the other hand, 7-O-acetyl-BFA, 4-O-acetyl-BFA, and 4,7-di-O-acetyl-BFA exhibited potential activities in cytotoxicity and inducibility of apoptosis. We suggested that the structural determinants for BFA include the moiety of the Michael acceptor, the conformational rigidity of the 13-membered ring, and the configuration of 4-hydroxyl group.

Functional morphology of antennal chemoreceptors of the parasitoid Microplitis croceipes (Hymenoptera: Braconidae).

The specialist parasitoid, Microplitis croceipes (Cresson) (Hymenoptera: Braconidae), uses chemical cues from plants damaged by herbivore-feeding and also plant by-products in host location and acceptance. These chemicals are detected by the wasp's antennae. We conducted scanning and transmission electron microscopy studies of male and female M. croceipes antennae and detected no distinct morphological differences in the chemoreceptors between the sexes. Male antennae are approximately twice as long as female antennae. We found five morphological sensillar types in both sexes: sensilla (s) trichodea were the most abundant and distributed over the whole antenna; s. chaetica were present in low numbers only on the scape and pedicel; and s. basiconica, s. coeloconica and elongated s. placodea were found only on the flagellum. Ultrastructural investigations revealed pore systems on s. basiconica and s. placodea. In s. placodea, sensory neurons run parallel to the longitudinal axis of the sensilla with terminal pores. We recorded responses from single olfactory receptor neurons in s. placodea to two plant-emitted volatiles, cis-3-hexenol and ocimene, and two anthropogenic compounds, cyclohexanone and 2-diisopropylaminoethanol. Male receptor neurons were more sensitive than those of females with significantly higher spike frequency being registered from neurons in males.