[Predictive value of ultrasound-guided anesthesia injection in arthroscopy for borderline developmental dysplasia of the hip].

Objective: To evaluate the value of ultrasound-guided intra-articular anesthetic injection in predicting postoperative outcomes for borderline developmental hip dysplasia (BDDH). Methods: A follow-up study. The clinical data of 37 BDDH patients who received ultrasound-guided intra-articular anesthetic injection and arthroscopic examination in the Department of Sports Medicine, Senior Department of Orthopedics, the Fourth Medical Center of PLA General Hospital from May 2018 to February 2021 were retrospectively analyzed. Among them, there were 17 males and 20 females with a mean age of (37.9±12.8) years. All patients underwent ultrasound-guided intra-articular anesthetic injection prior to arthroscopy, and were evaluated with hip physical examination before and after injection, as well as before and after arthroscopy, in order to obtain the visual analog score (VAS) of pain for seven assessments. The total VAS score was calculated based on these evaluations. Follow-up was conducted for at least 12 months. The effective rate of injection referred to the ratio of the improvement of VAS score after anesthetic injection to the total VAS score before injection. Pearson correlation analysis and Bland-Altman analysis were used to test the correlation between modified Harris hip score (mHHS) after ultrasound-guided intra-articular anesthetic injection and mHHS score after arthroscopic surgery. A binary logistic regression model was established to analyze the substantial clinical benefit (SCB) for patients. Following the logistic regression analysis, a receiver operating characteristic (ROC) curve was constructed to evaluate the predictive power of ultrasound-guided intra-articular anesthetic injection in achieving SCB in those patients. The optimal cut-off value for injection efficacy was determined based on the ROC curve when SCB was achieved. Results: The follow-up time for all patients was (26.3±7.6) months. After anesthetic injection for 20 minutes, the total VAS score of pain [M(Q1,Q3)] decreased from 13(8,23) points before injection to 1(0,4) points; and the mHHS score [M(Q1,Q3)] increased from 60(46,70) points before arthroscopy to 90(84,96) points after, with statistically significant differences before and after injection and before and after arthroscopy (both P<0.001). Pearson correlation analysis showed that the mHHS score after intra-articular anesthetic injection was positively correlated with the mHHS score after surgery (r=0.961, P<0.001). The area under the ROC curve for predicting SCB after arthroscopy with ultrasound-guided intra-articular anesthetic injection was 0.769 (95%CI: 0.561-0.976), the Youden index was 0.663, the cut-off value was 0.569 2, the sensitivity was 96.3%, and the specificity was 70.0%. Conclusions: The results of ultrasound-guided intra-articular anesthetic injection before arthroscopy can indicate the presence of intra-articular lesions, and the degree of pain relief after injection is proportional to the functional recovery after arthroscopy. Patients with intra-articular anesthetic injection efficacy>56.92% have better results in hip arthroscopy.

[Clinical efficacy of the treatment of bilateral gluteal muscle contracture by inside-out iliotibial band release under arthroscopy in the supine position].

Objective: To investigate the clinical efficacy of bilateral gluteal muscle contracture treated with inside-out iliotibial band release under arthroscopy in the supine position. Methods: A prospective non-randomized controlled trial. Forty-six patients admitted to the Department of Sports Medicine, Senior Department of Orthopedics, the Fourth Medical Center of PLA General Hospital from April 2021 to August 2022 for bilateral gluteal muscle contracture and proposed surgical treatment were enrolled. The subjects were divided into two groups according to the preferred surgical protocols of the patients: the supine position group was treated with inside-out iliotibial band release under arthroscopy in the supine position, and the operation in lateral position group was carried out with outside-in iliotibial band release under arthroscopy in the lateral position. The total duration of non-surgical operations and the total duration of surgical operations were recorded for all patients. The gluteal muscle contracture disability scale within 3 days before surgery and at least 2 months after surgery were compared between the two groups, and the occurrence of complications between the two groups was compared too. Results: There were 26 cases in the supine position group, 11 males and 15 females with a mean age of (31.8±7.3) years; and there were 20 cases in the lateral position group, 7 males and 13 females with a mean age of (30.6±6.3) years. The differences in gender, age, body mass index (BMI) and postoperative follow-up time between the two groups were not statistically significant (all P>0.05). The total duration of non-surgical operations was shorter in the supine position group than in the lateral position group [(47.9±10.4) min vs (63.9±7.5) min, P<0.001]. There was no statistically significant difference in the total duration of surgical operations between the supine position group and the lateral position group [31.0(27.0, 43.5) min vs 33.0(24.8, 38.0) min, P>0.05]. The postoperative gluteal muscle contracture disability scales were significantly improved in both the supine position and lateral position groups when compared with those before the operation [93.0 (85.0, 98.0) vs 61.0 (50.5, 66.8), P<0.001 and 88.5±6.9 vs 63.6±9.6, P<0.001, respectively]. There was no statistically significant difference in the gluteal muscle contracture disability scale between the supine position and lateral position groups before and 2 months after surgery [59.3±11.9 vs 63.6±9.6 and 93.0 (85.0, 98.0) vs 89.5(84.0, 94.8), both P>0.05, respectively]. Two patients in each group developed subcutaneous hematoma after surgery, and all of them resolved within 2 weeks after surgery, the difference in complication incidence rate was not statistically significant (P>0.05). No postoperative complications such as fat liquefaction in the operated area, infection, decreased hip abductor muscle strength or nerve injury in the lower extremity were observed in both groups. Conclusion: The treatment of bilateral gluteal muscle contracture by inside-out iliotibial band release under arthroscopy in the supine position can effectively improve clinical efficiency, with definite efficacy, and it is an operative program worth promoting.

[Outcomes of fluoroscopy-free hip arthroscopy technique in the treatment of femoroacetabular impingement syndrome].

Objective: To evaluate the effect of fluoroscopy-free technique in the arthroscopic treatment of femoroacetabular impingement syndrome (FAIS). Methods: A retrospective cohort study. Clinical data of FAI patients treated with hip arthroscopy in the No.4 Medical Center, PLA General Hospital from October 2018 to December 2021 were retrospectively analyzed. The patients were divided into two groups according to the surgical procedure: the fluoroscopy group and the fluoroscopy-free group. The operation time and modified Harris hip score (mHHS), international hip outcome tool (iHOT12) and visual analogue scale (VAS) of hip joint pain before and after the operation were observed and compared between the two groups. And the incidence of surgical complications in the two groups were compared too. Results: A total of 460 patients (213 males and 247 females) [aged (32.6±8.3) years (15-67 years)] with valid follow-up were included in this study. There were 275 cases in the fluoroscopy-free group and 185 cases in the fluoroscopy group. The operation time was shorter in the fluoroscopy-free group when compared with that in the fluoroscopy group, and the difference was statistically significant [(93.36±12.54) min vs (115.62±6.03) min, P<0.001]. In both groups, the VAS scores decreased and the mHHS scores and iHOT12 scores improved significantly at the last follow-up when compared with those before the operation (all P<0.001); however, there was no significant difference in the scores between the two groups (all P>0.05). The complication rate in the fluoroscopy-free group was 10.18% (28/275), and it was 10.81%(20/185) in the fluoroscopy group (P=0.829). Conclusion: Fluoroscopy-free hip arthroscopy technique for FAI can avoid radiation and shorten the operation time, but it does not increase the incidence of complications with reliable clinical outcomes.

[Risk factors and chemoprophylaxis of ulcerative colitis-colorectal cancer].

Ulcerative colitis-colorectal cancer (UC-CRC) is one of the most serious complications in patients with ulcerative colitis (UC), with worse prognosis and higher mortality than sporadic colorectal cancer (CRC). Since most UC-CRC developed through the "inflammation-dysplasia-carcinoma" approach, early detection of dysplasia through identification of high-risk groups reasonable monitoring and active prevention are extremely important. However, there is no consensus on the risk factors of UC carcinogenesis and the drugs that can be used for chemoprevention currently. This article combined with relevant literature at home and abroad, reviewed the current risk factors and chemopreventive drugs for UC carcinogenesis, in order to provide reference for early prevention, early detection and early diagnosis of UC-CRC.

[The expression of GGCT in the bladder urothelial cell carcinoma and its clinical significance].

Objective: To evaluate the expression and clinical significance of γ-glutamylcyclotransferase (GGCT) in patients with bladder urothelial cell carcinoma. Methods: Immunohistochemical staining for GGCT were performed on tissue sections of 86 patients with bladder urothelial cell carcinoma and 10 normal controls, and the correlations between GGCT and clinicopathological characteristics and the prognosis were analyzed. Results: The positive rate of the expression of GGCT in 86 cases of bladder urothelial cell carcinoma was 61.6% (53/86). GGCT protein was located mainly in cancer cell cytoplasm, and it can be seen in the nucleus of the tumor cells in some cases. The level of GGCT expression was positively related to pathological classification (P<0.001), stage (P=0.020), and tumor size (P=0.025). Immunohistochemical semiquantitative analysis showed that the expression of GGCT in patients with T1 stage of non-muscle invasion bladder urothelial cell carcinoma was significantly higher than that with Ta stage (P=0.034). Kaplan-Meier analysis showed that the expression of GGCT was correlated with the recurrence-free survival in patients with non-muscle invasive bladder cancer, the recurrence-free survival rate was lower in the GGCT positive group (P=0.029). Multivariate COX regression analysis showed that the pathological stage (OR=5.029, P=0.009) and the number of tumors (OR=3.320, P=0.024)were the independent risk factors for recurrence-free survival in patients with early urothelial cell carcinoma of the bladder. Conclusions: The expression of GGCT is significantly increased in bladder urothelial cell carcinoma and is related to the malignant biological behavior and progression of tumor. Patients with GGCT positive early bladder tumor are inclined to recur.

Seed-borne endophytic Bacillus velezensis LHSB1 mediate the biocontrol of peanut stem rot caused by Sclerotium rolfsii.

AIMS: This study aimed to obtain an antagonistic endophyte against Sclerotium rolfsii from peanut seeds, evaluate the biocontrol efficacy towards peanut stem rot and explore its antifungal mechanism against S. rolfsii. METHODS AND RESULTS: Thirty-seven endophytic bacteria were isolated from peanut seeds, six of which exhibited stronger antagonistic activities against S. rolfsii (inhibition rate, IR of hyphae growth ≥70%). Strain LHSB1, the strongest antagonistic strain, was identified as Bacillus velezensis. LHSB1 showed 93·8% of radial growth inhibition of S. rolfsii hyphae and exhibited obvious antagonistic activity against another six pathogenic fungi of peanut. Pot experiments showed two different LHSB1 treatments both significantly reduced the disease incidence and severity of stem rot (P < 0·05) compared to the controls, and the biocontrol efficacy reached 62·6-70·8%, significantly higher than that of Carbendazim control (P < 0·05). Further analyses revealed LHSB1 culture filtrate significantly inhibited sclerotia formation and germination, caused the abnormalities and membrane integrity damage of S. rolfsii hyphae, which might be the possible mode of action of LHSB1 against S. rolfsii. Three antifungal lipopeptides bacillomycin A, surfactin A and fengycin A, were detected in LHSB1 culture extracts by UPLC-ESI-MS, which could be responsible for the biocontrol activity of LHSB1 against S. rolfsii. CONCLUSION: Our results suggested that the seed-borne endophytic B. velezensis LHSB1 would be a tremendous potential agent for the biocontrol of peanut stem rot caused by S. rolfsii. SIGNIFICANCE AND IMPACT OF THE STUDY: This comprehensive study provides a candidate endophytic biocontrol strain and reveals its antifungal mechanism against S. rolfsi. To the best of our knowledge, this is the first time that seed-borne endophytic B. velezensis was used as the biocontrol agent to control peanut stem rot.

Knockout of a P-glycoprotein gene increases susceptibility to abamectin and emamectin benzoate in Spodoptera exigua.

P-glycoprotein [P-gp or the ATP-binding cassette transporter B1 (ABCB1)] is an important participant in multidrug resistance of cancer cells, yet the precise function of this arthropod transporter is unknown. The aim of this study was to determine the importance of P-gp for susceptibility to insecticides in the beet armyworm (Spodoptera exigua) using clustered regularly interspaced short palindromic repeats/CRISPR-associated 9 (CRISPR/Cas9) gene-editing technology. We cloned an open reading frame (ORF) encoding the S. exigua P-gp protein (SeP-gp) predicted to display structural characteristics common to P-gp and other insect ABCB1 transporters. A knockout line with a frame shift deletion of four nucleotides in the SeP-gp ORF was established using the CRISPR/Cas9 gene-editing system to test its potential role in determining susceptibility to chemical insecticides or insecticidal proteins from the bacterium Bacillus thuringiensis (Bt). Results from comparative bioassays demonstrate that knockout of SeP-gp significantly increases susceptibility of S. exigua by around threefold to abamectin and emamectin benzoate (EB), but not to spinosad, chlorfenapyr, beta-cypermethrin, carbosulfan indoxacarb, chlorpyrifos, phoxim, diafenthiuron, chlorfluazuron, chlorantraniliprole or two Bt toxins (Cry1Ca and Cry1Fa). Our data support an important role for SeP-gp in susceptibility of S. exigua to abamectin and EB and imply that overexpression of SeP-gp may contribute to abamectin and EB resistance in S. exigua.

[Dynamic investigation of nutritional risk in patients with malignant tumor during hospitalization].

Objective: To prospectively investigate the changes in nutritional status of patients with malignant tumors during hospitalization by using nutritional risk screening (NRS2002), and to analyze the correlation between the nutritional status and clinical outcomes . Methods: This was a prospective and parallel research done by multi-center collaboration from 34 hospitals in China from June to September 2014.Hospitalized patients with malignant tumors inthese departments (Department of Gastroenterology, respiratory medicine, oncology, general surgery, thoracic surgery and geriatrics)were investigated. Only the patients with age≥ 18 years and hospitalization time between 7-30 days were included. During hospitalization, the physical indexes of human bodywere measured, and the NRS 2002 scores, and monitored the nutritional support at the time points of admission and 24 hours before discharge were recorded.And whether there was a nutritional risk in hospitalized patients and its association with clinical outcomes were investigated. Results: A total of 2 402 patients with malignancies were enrolled in this study. Seventy fourpatients who did not complete NRS2002 were eliminated, and 2 328 patients were included. The number of the main diseases was the top five, including 587 cases of colorectal cancer, 567 cases of lung cancer, 564 cases of gastric cancer, 146 cases of esophageal cancer, and 119 cases of liver tumor. At the time of discharge, compared with admission, the BMI, body weight, grip and calf circumferences of patients with malignant tumor were significantly decreased (P<0.05). The total protein, albumin, prealbumin and hemoglobin were significantly lower than those at admission (P<0.05). In 2 328 patients who were completed nutritional risk screening, the rate of malnutrition at admission was 11.1% (BMI =18.5, 258/2 328) and the rate of malnutrition at discharge was 10.9% (BMI =18.5, 254/2 328), there were no significant differences (χ(2)=0.019 7, P=0.888). There were 1 204 patients with nutritional risk at admission (51.7%, NRS2002 score≥3)and 1 352 patients with nutritional risk at discharge (58.1%, NRS2002 score≥3), with significant differences (χ(2)=49.9, P<0.001). The incidence of nutritional risk in patients with colorectal, stomach, and lung tumors at discharge was significantly higher than that at admission (P<0.05). The infective complications and other complications of patients with nutritional risk were significantly greater than those without nutritional risk at admission and at discharge.ICU hospitalization stay of patients with nutritional risk was increased significantly than those without nutritional risk at admission(P=0.042). Hospitalization expenses of patients with nutritional risk was increased significantly than those of patients without nutritional risk at discharge(P<0.01). Conclusion: The patients with malignant tumor have a higher incidence rate of malnutrition at both admission and discharge and malnutritionhas correlation with adverse clinical outcomes.The aboveindicators did not improve significantly at discharge.Doctors should pay more attention to the nutritional status (screening and evaluation)of patients before discharge and use appropriate and adequate nutrition support in order to prevent the weight loss and improve the life quality of patients.

[Clinical study of no necrotic cavity lavage after debridement and drainage in patients with infected pancreatic necrosis].

Objective: To observe the outcomes of no necrotic cavity lavage after debridement and drainage in patients with infected pancreatic necrosis(IPN). Methods: From February 2014 to August 2017, there were 89 patients who were diagnosed as IPN undergoing minimally invasive surgery with no necrotic cavity lavage and large caliber-wide channel drainage in Department of General Surgery, Xuanwu Hospital, Capital Medical University. There were 57 male and 32 female patients aging of (49.5±14.4)years (ranging from 23 to 84 years). The body mass index of 89 patients was (25.4±3.8)kg/m(2) (ranging from 17.6 to 36.7 kg/m(2)). Among the 89 patients, 37 cases(41.6%) of biliary pancreatitis, 10 cases (11.2%) of alcoholic pancreatitis, 16 cases(18.0%) of hyperlipidemic pancreatitis, and 26 cases(29.2%) of other reasons. Results: Of 89 patients, IPN in 6 patients(6.7%) resolved using only percutaneous catheter drainage; another 83 patients underwent laparoscopic debridement(n=3, 3.4%) or video-assisted debridement(n=80, 89.9%). No patient was conversed to laparotomy. The average operation frequency and surgery time was (2.3±1.7) times and (56.5±31.7) minutes.The median bleeding volume and total length of stay was 10(0-600) ml and 34(6-172) days separately. The complication rate(Clavien-Dindo grade≥Ⅲ) was 9.0%(8/89) which involved mainly abdominal hemorrhage (5/8) and digestive tract fistula formation (3/8). The overall mortality rate was 6.7%(6/89). Among them, 3 cases died of abdominal infection, bacteremia and multiple organ failure, 2 cases died of pulmonary infection and bacteremia and 1 case died of fungal infection. Conclusion: No necrotic cavity lavage after debridement and drainage operation is considered effective and safe for IPN patients.

[Research Progress of the Relationship between SUNDS and OSAHS].

Sudden unexplained nocturnal death syndrome （SUNDS） is always a difficulty in forensic medicine researches. Although the development of molecular genetics promotes the etiologic study of SUNDS, the pathogenesis of most such cases is still unclear. Sleep apnea syndrome （SAS） is one of the common forms of sleep disorders, and obstructive sleep apnea hypopnea syndrome （OSAHS） is the most common. In recent years, some domestic and international researches show that OSAHS is related to the development of cardiovascular disease, which may cause cardiac arrhythmia, even sudden death. This article reviews the relationship between SUNDS and OSAHS and aims to provide new ideas for the pathogenesis of SUNDS.

[Association of sleep duration and physical exercise with dyslipidemia in older adults aged 80 years and over in China].

Objective: To explore the impact of sleep duration, physical exercise, and their interactions on the risk of dyslipidemia in older adults aged ≥80 (the oldest old) in China. Methods: The study subjects were the oldest old from four rounds of Healthy Aging and Biomarkers Cohort Study (2008-2009, 2011-2012, 2014 and 2017-2018). The information about their demographic characteristics, lifestyles, physical examination results and others were collected, and fasting venous blood samples were collected from them for blood lipid testing. Competing risk model was used to analyze the causal associations of sleep duration and physical exercise with the risk for dyslipidemia. Restricted cubic spline (RCS) function was used to explore the dose-response relationship between sleep duration and the risk for dyslipidemia. Additive and multiplicative interaction model were used to explore the interaction of sleep duration and physical exercise on the risk for dyslipidemia. Results: The average age of 1 809 subjects was (93.1±7.7) years, 65.1% of them were women. The average sleep duration of the subjects was (8.0±2.5) hours/day, 28.1% of them had sleep duration for less than 7 hours/day, and 27.2% had sleep for duration more than 9 hours/day at baseline survey. During the 9-year cumulative follow-up of 6 150.6 person years (follow-up of average 3.4 years for one person), there were 304 new cases of dyslipidemia, with an incidence density of 4 942.6/100 000 person years. The results of competitive risk model analysis showed that compared with those who slept for 7-9 hours/day, the risk for dyslipidemia in oldest old with sleep duration >9 hours/day increased by 22% (HR=1.22, 95%CI: 1.07-1.39). Compared with the oldest old having no physical exercise, the risk for dyslipidemia in the oldest old having physical exercise decreased by 33% (HR=0.67, 95%CI: 0.57-0.78). The RCS function showed a linear positive dose-response relationship between sleep duration and the risk for hyperlipidemia. The interaction analysis showed that physical exercise and sleep duration had an antagonistic effect on the risk for hyperlipidemia. Conclusion: Physical exercise could reduce the adverse effects of prolonged sleep on blood lipids in the oldest old.

Effects of fluoride and chloride on the growth of Chlorella pyrenoidosa.

To compare the toxic action of fluoride (F(-)) and chloride (Cl(-)) on freshwater algae, effects of F(-) and Cl(-) on Chlorella pyrenoidosa growth were investigated by determination of the algal biomass and model analysis. Results showed that the exponential growth equation (D = D0 + ae(bT)) may be used to fit the relationship between cell density (D) of C. pyrenoidosa and exposure time (T), but F(-) and Cl(-) have significantly different effects on parameters a (initial quantity of algae at the exponential growth stage) and b (growth constant). In the range of experimental concentrations, F(-) inhibited C. pyrenoidosa growth, and percent inhibition increased with increasing exposure time and F(-) concentration. By contrast, Cl(-) either inhibited or enhanced C. pyrenoidosa growth depending on Cl(-) concentration and exposure time, and percent inhibition increased with increasing Cl(-) concentration but decreased with increasing exposure time. The dose-response equation: I = min + (max - min)/(1 + 10(lgEC50-lgC)) may be used to fit the relationship between percent inhibition (I) and F(-) or Cl(-) concentration (C); however, F(-) was distinct from Cl(-) in terms of effects on parameter 'min'. The EC50 values of F(-) and Cl(-) ranged from 118 to 170 mg/L and 19.73 to 36.33 g/L, respectively; the latter is much higher than the former. Thus F(-) has higher toxicity than Cl(-).

[From treatment to whole course management: envisioning comprehensive management of Talaromycosis marneffei].

Talaromycosis marneffei has been increasing in recent years. Our understanding of this disease has gradually deepened through extensive basic and clinical research, but there are still many limitations. In this article, by incorporating the latest research advancements, we discuss important issues in managing Talaromycosis marneffei trends, aiming to guide effective prevention and control of the disease, improving public health, and reducing the healthcare burden.

SD0006 promotes nucleus pulposus cell proliferation via the p38MAPK/HDAC4 pathway.

OBJECTIVE: p38 mitogen-activated protein kinase (p38MAPK) is a critical negative regulator for nucleus pulposus (NP) cell metabolism contributing to intervertebral disc degeneration (IDD). Histone deacetylase 4 (HDAC4) has the ability to mediate cell proliferation and is affected by p38MAPK. It is unclear whether the p38 MAPK inhibitor SD0006 (SD) can regulate IDD. Our study aims to explore the effect of SD in the progression of IDD, as well as its potential mechanism. PATIENTS AND METHODS: NP cells isolating from mild degenerated NP tissues were cultured, and IL-1ß or Asiatic acid (AA) was used to activate p38MAPK and accelerate the NP cell degradation. Then, SD was used to reject the p38MAPK activation. After that, the levels of phosphorylated p38MAPK (p-p38), HDAC4, proliferating cell nuclear antigen (PCNA), inflammatory factor, proliferative cell rate, and cell cycle were determined by Western blot, RT-PCR, flow cytometry, and immunofluorescence, respectively. RESULTS: The results showed that the activation of p38MAPK by IL-1ß and AA decreased the HDAC4 expression, affected the collagen-Ⅱ expression, upregulated the TNF-α, IL-6, MMP-3, and ADAMTS mRNA levels, and prevent the NP cell proliferation by mediating cell cycles. However, the application of SD alleviated the negative effect of p-p38 by upregulating HDAC4, anti-inflammation, and promoting cell proliferation, while the blocking of HDAC4 expression partly abolished the effect of SD. CONCLUSIONS: SD can prevent NP cell degeneration by promoting cell proliferation and suppressing inflammation via p38MAPK/HDAC4 pathway.

Theory and modeling of stereoselective organic reactions.

Theoretical investigations of the transition structures of additions and cycloadditions reveal details about the geometries of bond-forming processes that are not directly accessible by experiment. The conformational analysis of transition states has been developed from theoretical generalizations about the preferred angle of attack by reagents on multiple bonds and predictions of conformations with respect to partially formed bonds. Qualitative rules for the prediction of the stereochemistries of organic reactions have been devised, and semi-empirical computational models have also been developed to predict the stereoselectivities of reactions of large organic molecules, such as nucleophilic additions to carbonyls, electrophilic hydroborations and cycloadditions, and intramolecular radical additions and cycloadditions.

Effects of cyclooxygenase-2 non-selective and selective inhibitors on proliferation inhibition and apoptosis induction of esophageal squamous carcinoma cells.

The objective of this study is to investigate the effects of aspirin and nimesulide on cell proliferation, apoptosis and its potential mechanisms in EC9706 and EC109 esophageal squamous carcinoma cells. EC9706 and EC109 cells were incubated with varying concentrations of aspirin and nimesulide, and the effects on cell proliferation and apoptosis were monitored by 3-(4,5-dimethyl-thiazol-2yl)-2,5-diphenyltetrazolium bromide and flow cytometry. Reverse transcription-polymerase chain reaction and Western blot assays were used to investigate expression of Bcl-2 and Bax. Prostaglandin E2 production was measured by enzyme linked immunosorbent assay. Pretreatment with aspirin and nimesulide inhibited EC9706 and EC109 cell growth in a time and dose-dependent manner, accompanied with a decrease of prostaglandin E2 production. In EC9706 cells, the mechanism of aspirin and nimesulide induced growth inhibition was a consequence of cell cycle arrest at the G(0)/G(1) check point. In EC109 cells, growth arrest was by induction of apoptosis, associated with downregulation of Bcl-2, but not Bax. In conclusion, aspirin and nimesulide could inhibit cell proliferation and induce apoptosis in esophageal squamous carcinoma cells. Cyclooxygenase-2 inhibitor may be a promising therapeutic agent for human esophageal squamous cell carcinoma.

Introgression of a disrupted cadherin gene enables susceptible Helicoverpa armigera to obtain resistance to Bacillus thuringiensis toxin Cry1Ac.

A disrupted allele (r1) of a cadherin gene (Ha_BtR) is genetically associated with incompletely recessive resistance to Bacillus thuringiensis toxin Cry1Ac in a Cry1Ac-selected strain (GYBT) of Helicoverpa armigera. The r1 allele of Ha_BtR was introgressed into a susceptible SCD strain by crossing the GYBT strain to the SCD strain, followed by repeated backcrossing to the SCD strain and molecular marker assisted family selection. The introgressed strain (designated as SCD-r1, carrying homozygous r1 allele) obtained 438-fold resistance to Cry1Ac, >41-fold resistance to Cry1Aa and 31-fold resistance Cry1Ab compared with the SCD strain; however, there was no significant difference in susceptibility to Cry2Aa between the integrated and parent strains. It confirms that the loss of function mutation of Ha_BtR alone can confer medium to high levels of resistance to the three Cry1A toxins in H. armigera. Reciprocal crosses between the SCD and SCD-r1 strains showed that resistance to Cry1Ac in the SCD-r1 strain was completely recessive. Life tables of the SCD and SCD-r1 strains on artificial diet in the laboratory were constructed, and results showed that the net replacement rate (R0) did not differ between the strains. The toxicity of two chemical insecticides, fenvalerate and monocrotophos, against the SCD-r1 strain was not significantly different from that to the SCD strain. However, larval development time of the SCD-r1 strain was significantly longer than that of the SCD strain, indicating a fitness cost of slower larval growth is associated with Ha_BtR disruption in H. armigera.

Assessing the susceptibility of cruciferous Lepidoptera to Cry1Ba2 and Cry1Ca4 for future transgenic cruciferous vegetables.

Advances in transgenic plants expressing Bacillus thuringiensis (Bt) insecticidal gene(s) offer a promising alternative to traditional insecticides for control of lepidopteran pests on important cruciferous vegetable crops such as cabbage and cauliflower. A public-private partnership, the Collaboration on Insect Management for Brassicas in Asia and Africa (CIMBAA), was formed in 2005 with the goal of developing dual-gene Bt cauliflower and cabbage, initially for India, to replace the use of broad spectrum, traditional insecticides. As a first step in this effort, the major lepidopteran pests of cruciferous vegetable crops [Plutella xylostella (L.), Pieris rapae (L.), Pieris brassicae (L.), Crocidolomia binotalis (L.), Hellula undalis (F.), Diacrisia obliqua Walker, Spodoptera litura F., and Helicoverpa armigera (Hübner)] were collected over a large geographic area (India, Indonesia, Taiwan, China, Australia, and the United States) and tested against purified Cry1Ba2 and Cry1Ca4 toxins, the toxins proposed to be expressed in the CIMBAA plants. Our results demonstrate that Cry1Ba2 and Cry1Ca4 were effective against the primary target of the CIMBAA plants, P. xylostella, regardless of geographic location, and had LC50 values <1.3 ppm. Furthermore, one or both toxins were effective against the other major pest Lepidoptera, except for S. litura or H. armigera which were less susceptible. No cross-resistance has been found between Cry1Ba2 and Cry1Ca4, suggesting cry1Ba2+cry1Ca4 cauliflower and cabbage could be an effective and sustainable tool to control, P. xylostella, the key lepidopteran pest on cruciferous vegetable crops, as well as most other Lepidoptera. As the CIMBAA plants are being developed, further tests are needed to determine whether they will express these proteins at sufficient levels to control all the Lepidoptera. Sustainable use of the dual-gene plants also is discussed.