Development and economic assessment of machine learning models to predict glycosylated hemoglobin in type 2 diabetes.

Background: Glycosylated hemoglobin (HbA1c) is recommended for diagnosing and monitoring type 2 diabetes. However, the monitoring frequency in real-world applications has not yet reached the recommended frequency in the guidelines. Developing machine learning models to screen patients with poor glycemic control in patients with T2D could optimize management and decrease medical service costs. Methods: This study was carried out on patients with T2D who were examined for HbA1c at the Sichuan Provincial People's Hospital from April 2018 to December 2019. Characteristics were extracted from interviews and electronic medical records. The data (excluded FBG or included FBG) were randomly divided into a training dataset and a test dataset with a radio of 8:2 after data pre-processing. Four imputing methods, four screening methods, and six machine learning algorithms were used to optimize data and develop models. Models were compared on the basis of predictive performance metrics, especially on the model benefit (MB, a confusion matrix combined with economic burden associated with therapeutic inertia). The contributions of features were interpreted using SHapley Additive exPlanation (SHAP). Finally, we validated the sample size on the best model. Results: The study included 980 patients with T2D, of whom 513 (52.3%) were defined as positive (need to perform the HbA1c test). The results indicated that the model trained in the data (included FBG) presented better forecast performance than the models that excluded the FBG value. The best model used modified random forest as the imputation method, ElasticNet as the feature screening method, and the LightGBM algorithms and had the best performance. The MB, AUC, and AUPRC of the best model, among a total of 192 trained models, were 43475.750 (¥), 0.972, 0.944, and 0.974, respectively. The FBG values, previous HbA1c values, having a rational and reasonable diet, health status scores, type of manufacturers of metformin, interval of measurement, EQ-5D scores, occupational status, and age were the most significant contributors to the prediction model. Conclusion: We found that MB could be an indicator to evaluate the model prediction performance. The proposed model performed well in identifying patients with T2D who need to undergo the HbA1c test and could help improve individualized T2D management.

A personalized prediction model for urinary tract infections in type 2 diabetes mellitus using machine learning.

Patients with type 2 diabetes mellitus (T2DM) are at higher risk for urinary tract infections (UTIs), which greatly impacts their quality of life. Developing a risk prediction model to identify high-risk patients for UTIs in those with T2DM and assisting clinical decision-making can help reduce the incidence of UTIs in T2DM patients. To construct the predictive model, potential relevant variables were first selected from the reference literature, and then data was extracted from the Hospital Information System (HIS) of the Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital for analysis. The data set was split into a training set and a test set in an 8:2 ratio. To handle the data and establish risk warning models, four imputation methods, four balancing methods, three feature screening methods, and eighteen machine learning algorithms were employed. A 10-fold cross-validation technique was applied to internally validate the training set, while the bootstrap method was used for external validation in the test set. The area under the receiver operating characteristic curve (AUC) and decision curve analysis (DCA) were used to evaluate the performance of the models. The contributions of features were interpreted using the SHapley Additive ExPlanation (SHAP) approach. And a web-based prediction platform for UTIs in T2DM was constructed by Flask framework. Finally, 106 variables were identified for analysis from a total of 119 literature sources, and 1340 patients were included in the study. After comprehensive data preprocessing, a total of 48 datasets were generated, and 864 risk warning models were constructed based on various balancing methods, feature selection techniques, and a range of machine learning algorithms. The receiver operating characteristic (ROC) curves were used to assess the performances of these models, and the best model achieved an impressive AUC of 0.9789 upon external validation. Notably, the most critical factors contributing to UTIs in T2DM patients were found to be UTIs-related inflammatory markers, medication use, mainly SGLT2 inhibitors, severity of comorbidities, blood routine indicators, as well as other factors such as length of hospital stay and estimated glomerular filtration rate (eGFR). Furthermore, the SHAP method was utilized to interpret the contribution of each feature to the model. And based on the optimal predictive model a user-friendly prediction platform for UTIs in T2DM was built to assist clinicians in making clinical decisions. The machine learning model-based prediction system developed in this study exhibited favorable predictive ability and promising clinical utility. The web-based prediction platform, combined with the professional judgment of clinicians, can assist to make better clinical decisions.

Develop an ADR prediction system of Chinese herbal injections containing Panax notoginseng saponin: a nested case-control study using machine learning.

OBJECTIVE: This study aimed to develop an adverse drug reactions (ADR) antecedent prediction system using machine learning algorithms to provide the reference for security usage of Chinese herbal injections containing Panax notoginseng saponin in clinical practice. DESIGN: A nested case-control study. SETTING: National Center for ADR Monitoring and the Electronic Medical Record (EMR) system. PARTICIPANTS: All patients were from five medical institutions in Sichuan Province from January 2010 to December 2018. MAIN OUTCOMES/MEASURES: Data of patients with ADR who used Chinese herbal injections containing Panax notoginseng saponin were collected from the National Center for ADR Monitoring. A nested case-control study was used to randomly match patients without ADR from the EMR system by the ratio of 1:4. Eighteen machine learning algorithms were applied for the development of ADR prediction models. Area under curve (AUC), accuracy, precision, recall rate and F1 value were used to evaluate the predictive performance of the model. An ADR prediction system was established by the best model selected from the 1080 models. RESULTS: A total of 530 patients from five medical institutions were included, and 1080 ADR prediction models were developed. Among these models, the AUC of the best capable one was 0.9141 and the accuracy was 0.8947. According to the best model, a prediction system, which can provide early identification of patients at risk for the ADR of Panax notoginseng saponin, has been established. CONCLUSION: The prediction system developed based on the machine learning model in this study had good predictive performance and potential clinical application.

Predictive models of medication non-adherence risks of patients with T2D based on multiple machine learning algorithms.

OBJECTIVE: Medication adherence plays a key role in type 2 diabetes (T2D) care. Identifying patients with high risks of non-compliance helps individualized management, especially for China, where medical resources are relatively insufficient. However, models with good predictive capabilities have not been studied. This study aims to assess multiple machine learning algorithms and screen out a model that can be used to predict patients' non-adherence risks. METHODS: A real-world registration study was conducted at Sichuan Provincial People's Hospital from 1 April 2018 to 30 March 2019. Data of patients with T2D on demographics, disease and treatment, diet and exercise, mental status, and treatment adherence were obtained by face-to-face questionnaires. The medication possession ratio was used to evaluate patients' medication adherence status. Fourteen machine learning algorithms were applied for modeling, including Bayesian network, Neural Net, support vector machine, and so on, and balanced sampling, data imputation, binning, and methods of feature selection were evaluated by the area under the receiver operating characteristic curve (AUC). We use two-way cross-validation to ensure the accuracy of model evaluation, and we performed a posteriori test on the sample size based on the trend of AUC as the sample size increase. RESULTS: A total of 401 patients out of 630 candidates were investigated, of which 85 were evaluated as poor adherence (21.20%). A total of 16 variables were selected as potential variables for modeling, and 300 models were built based on 30 machine learning algorithms. Among these algorithms, the AUC of the best capable one was 0.866±0.082. Imputing, oversampling and larger sample size will help improve predictive ability. CONCLUSIONS: An accurate and sensitive adherence prediction model based on real-world registration data was established after evaluating data filling, balanced sampling, and so on, which may provide a technical tool for individualized diabetes care.

Nobiletin protects human retinal pigment epithelial cells from hydrogen peroxide-induced oxidative damage.

Nobiletin (3',4',5,6,7,8-hexamethoxyflavone), a dietary polymethoxylated flavonoid found in Citrus fruits, has been reported to have antioxidant effect. However, the effect of nobiletin on human retinal pigment epithelium (RPE) cells induced by hydrogen peroxide (H2 O2 ) is still unclear. Therefore, we investigated the protective effect of nobiletin against H2 O2 -induced cell death in RPE cells. Our results demonstrated that nobiletin significantly increased cell viability from oxidative stress. Nobiletin inhibited H2 O2 -induced ROS production and caspase-3/7 activity in ARPE-19 cells. Furthermore, nobiletin significantly increased Akt phosphorylation in ARPE-19 cells exposed to H2 O2 . Meanwhile, LY294002, an inhibitor of PI3K/Akt, abolished the protective effect of nobiletin against H2 O2 -induced decreased cell viability and increased caspase-3/7 activity in ARPE-19 cells. In summary, these data show that nobiletin protects RPE cells against oxidative stress through activation of the Akt-signaling pathway. Thus, nobiletin should be an oxidant that attenuates the development of age-related macular degeneration.

Clinical efficacy of integrative therapy in the treatment of non - proliferative diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

·AIM: To study the clinical efficacy of integrative therapy in the treatment of non-proliferative diabetic retinopathy.·METHODS: Ninety patients ( 90 eyes ) in our hospital with non - proliferative diabetic retinopathy were randomly divided into three groups. All three groups were treated with diabetes drugs to control blood sugar. The first group was treated with western medicine, the second group was treated with Chinese medicine decoction Traditional Chinese Medicine ( TCM ) treatment, and the third group was treated with the combination of those two methods. All patients were recorded and analyzed changes of clinical effects after 6 courses of treatment.·RESULTS: After 6 courses of treatment, the total efficacy rate of the third group was 86%, markedly higher than that of the first group (57%, P<0. 05) as well as the second group (60%, P<0. 05). The integrative group improved more markedly in terms of vision, macular edema absorption, and ERG b-wave amplitude restoration, the difference being statistically significant (P<0. 05) when compared to the first and the second group.· CONCLUSION: Integrative treatment of diabetic retinopathy could effectively improve the therapeutic effect in patients with non-proliferative retinopathy.

Lutein supplementation improves visual performance in Chinese drivers: 1-year randomized, double-blind, placebo-controlled study.

OBJECTIVES: Although it is known that the carotenoid lutein can affect visual performance, we still have much to learn about its effect in occupational populations, like drivers. The aim of this study was to examine the effect of lutein supplementation on visual function in healthy drivers with long-term light exposure. METHODS: The study was a randomized, double-blind, placebo-controlled, 1-y intervention study. It included 120 normal participants (drivers). The active (A) group consumed 20 mg of lutein daily. Participants were assessed at baseline, 1, 3, 6, and 12 mo (V0, V1, V2, V3, and V4, respectively). Assessment included visual acuity, serum lutein concentrations, macular pigment optical density (MPOD), and visual performance. At the onset and at the end of the intervention, dietary intakes of lutein and visual-related quality of life were measured. RESULTS: There was a trend (in the active group) toward an increase in best spectacle-corrected visual acuity measured, but there were no significant differences. Serum lutein and central MPOD in the active group increased significantly, whereas no change was observed in the placebo group. We observed significant increases in contrast and glare sensitivity, especially in the mesopic condition. There were significant improvements in the score of the National Eye Institute 25-Item Visual Functioning Questionnaire driving subscale in the active group. CONCLUSIONS: Daily supplementation with 20 mg of lutein increases MPOD levels. Lutein may benefit driving at night and other spatial discrimination tasks carried out under low illumination.

[Protective effect of valproic acid on ischemia-reperfusion induced injury in retina of rat].

OBJECTIVE: To study the protective effects and the related mechanism of valproic acid (VPA) on ischemia-reperfusion-induced injury in retina of rat. METHODS: Experimental study. Ninety Wistar rats were divided randomly into three groups: normal (blank) control group, retinal ischemia-reperfusion (experimental control) group treated with PBS, retinal ischemia-reperfusion (experimental) group treated with VPA. Retinal ischemia was induced by acute high intraocular pressure. Rats were executed at 6, 12, 24, 48 h and 7 d after reperfusion. The eyeballs were enucleated for retinal histopathological examination. The fluoro-gold retrograde labeling was performed and the survival of retinal ganglion cells was analyzed by calculating the densities in fluoro-gold labeled retinal ganglion cell. The protein expression of heat shock protein 70 (Hsp70) and the acetylation of transcription factor Sp1 were analyzed by Western blot assay. The levels of bcl-2 and bax mRNA were analyzed by RT-PCR assay. To determine the significance of differences, analysis of paired-samples t-test was carried out. RESULTS: (1) The HE staining of retinal histological section showed that the edema of retina were attenuated significantly by VPA in experimental group, the difference between the experimental group and the experimental control group was statistically significant (t = 7.491, P < 0.05). The fluoro-gold retrograde labeling showed that the survival of retinal ganglion cells in experimental group [(1629 ± 63)/mm(2)] was significantly higher than experimental group [(908 ± 65)/mm(2)], the difference between the two groups was statistically significant (t = 7.248, P < 0.05). (2) The immuno-blot analysis showed that VPA resulted in significant increase in expression of Hsp70 protein in ischemic retinas, the difference between experimental group and experimental control group was statistically significant (t = 6.176, P < 0.05). The acetylation of Sp1 was significantly higher in experimental group than experimental control group, the difference between the two groups was statistically significant (t = 11.264, P < 0.05). The RT-PCR analysis showed that VPA increased the expression of bcl-2 mRNA in experimental group (0.403 ± 0.009), the difference between experimental group and experimental control group (0.314 ± 0.012) was statistically significant (t = 5.489, P < 0.05). VPA attenuated the expression of bax mRNA in experimental group (0.383 ± 0.009), the difference between experimental group and experimental control group (0.492 ± 0.016) was statistically significant (t = 5.723, P < 0.05). CONCLUSIONS: (1) VPA protected retina from ischemic injury. (2) The upregulation of Hsp70 and bcl-2, downregulation of bax maybe involved in the mechanism of the protection.

Effects of estrogen on collagen gel contraction by human retinal glial cells.

BACKGROUND: There are definite gender differences in patients with macular holes. Menopausal women over 50 years are most affected. We aimed to observe the effect of estrogen on collagen gel contraction by cultured human retinal glial cells. It is speculated that estrogen could strengthen the tensile stress of the macula by maintaining the correct morphology and contraction. METHODS: Estrogen was used to determine its effects on collagen gel contraction, and its function was measured using morphological changes in cells. Human retinal glial cells were cultured in collagen solution. The cells were then exposed to collagen gels and the degree of contraction of the gel was determined. RESULTS: Estrogen at differing concentrations had no effect on the growth of human retinal glial cells. However, after exposed to collagen gel block, less contraction was noted in the estrogen-treated group than in the control group. CONCLUSIONS: Estrogen can inhibit collagen gel contraction by glial cells. These results suggest a mechanism for macular hole formation, which is observed in menopausal females.

An experimental study of a modified dahuang zhechong pill on the--angiogenesis of RF/6A cells in vitro.

OBJECTIVE: To investigate the effects of a modified Dahuang Zhechong Pill (MDZP) on the angiogenesis of rhesus choroid-retina endothelial (RF/6A) cells and its preliminary mechanism. METHODS: A 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) method was used to assess the effect of a MDZP on RF/6A cell proliferation induced by vascular endothelial growth factor (VEGF). Transwell inserts were used to assess the effect of the MDZP on RF/6A cell migration. Matrigel was used to assess the effect of the MDZP on the tube formation of RF/ 6A cells. Western blotting and quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) were used to detect the protein and mRNA expression, respectively, of VEGF and matrix metalloproteinase-2 (MMP-2) in RF/6A cells treated with the MDZP. RESULTS: RF/6A cell proliferation induced by VEGF was inhibited by 0.2 mg/mL MDZP. At 0, 12.5, 25 and 50 mg/mL MDZP, the number of cells that migrated through Transwell membranes was 73.33 +/- 4.51, 61.33 +/- 4.04, 28.67 +/- 6.66 and 17.67 +/- 4.16, respectively, and the number of tubes formed in Matrigel was 20.33 +/- 0.58, 13.33 +/- 1.53, 11.00 +/- 1.00 and 1.33 +/- 0.58, respectively. At 100 and 200 mg/mL MDZP, the protein and mRNA expression of VEGF and MMP-2 were inhibited in RF/6A cells. At 400 mg/mL MDZP, the expression of VEGF mRNA and MMP-2 protein were inhibited in RF/6A cells. CONCLUSIONS: MDZP inhibits the angiogenesis of RF/6A cells via the suppression of proliferation, migration and tube formation of RF/6A cells. Inhibition of the protein and mRNA expression of VEGF and MMP-2 in RF/6A cells may be an important mechanism.

[Chinese herbal medicine Lingqi Huangban Granule protects retinal pigment epithelial cells against oxidative stress-induced injury in vitro].

OBJECTIVE: To observe the protective effects of drug-contained serum of Lingqi Huangban Granule (LQHBG), a compound traditional Chinese herbal medicine, on oxidative stress-induced injury in rabbit retinal pigment epithelial (RPE) cells in vitro. METHODS: The oxidative stress of rabbit RPE cells in vitro was induced with hydrogen peroxide (500µmol/L) and different concentrations of LQHBG were administered to rats to prepare medicated serum. RPE cells were randomized into normal control group (no hydrogen peroxide), model group (hydrogen peroxide), model plus serum group (hydrogen peroxide and 10% control serum), model plus low-dose LQHBG group (hydrogen peroxide and low-dose LQHBG-medicated serum) and model plus high-dose LQHBG group (hydrogen peroxide and high-dose LQHBG-medicated serum). Teminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay and flow cytometry (FCM) were used to measure apoptosis of cultured rabbit RPE cells. Protein expressions of caspase-3 and Bcl-X(L) were observed by Western blot method. RESULTS: FCM results showed that the apoptotic rates of the normal control group, model group, control serum group and serum containing low- and high-dose LQHBG groups were (4.85±0.26)%, (20.02±1.37)%, (21.84±0.94)%, (13.56±0.55)%, and (8.58±0.39)%, respectively; compared with the model group, the apoptotic rates of RPE cells in the low- and high-dose LQHBG groups were obviously reduced in a dose-related manner (P<0.05). TUNEL results showed that nuclei of apoptotic cells were stained brown; the number of apoptotic cells in the low- and high-dose LQHBG groups was obviously less than that in the model group. The protein expression of caspase-3 was up-regulated in the model and control serum groups, which was higher than that in the high-dose LQHBG group (P<0.05). The protein expression of Bcl-X(L) was down-regulated in the model and control serum groups, which was lower than that in the low- and high-dose LQHBG groups (P<0.01). CONCLUSION: Drug-contained serum of LQHBG obviously reduces apoptosis and partly protects rabbit RPE cells from oxidative stress-induced injury. The protective function is due to an improvement in antioxidant abilities, down-regulation of the expression of caspase-3 and up-regulation of the expression of Bcl-X(L).

[Protective effects of Ginkgo biloba extract on morphology and function of retinal ganglion cells after optic nerve transection in guinea pigs].

OBJECTIVE: To investigate the effects of Ginkgo biloba extract (EGb 761) on the morphology and function of retinal ganglion cells (RGC) in guinea pigs with optic nerve transection. METHODS: Seventy-five albino guinea pigs were randomly divided into five groups: normal control group, sham-operated group, untreated group, normal saline group and EGb 761 group. No operation was performed in the normal control group. Optic nerve was merely exposed in the sham-operated group, but transected at 1.0 mm from posterior pole of the eye ball in the untreated, normal saline and EGb 761 groups. Guinea pigs in the EGb 761 group or the normal saline group received daily intraperitoneal injection of EGb 761 (100 mg/kg) or corresponding volume of normal saline from 7 days before experiment to 28 days after experiment. Three guinea pigs in each group were sacrificed for apoptosis assay (TUNEL method) of RGC. Pattern electoretinograms (PERGs) were recorded 14 and 28 days after transection, respectively. At the end of the examination, six guinea pigs were killed for histological examination and RGC count. RESULTS: No TUNEL-positive cells were observed in the normal control, sham-operated and EGb 761 groups, but there were TUNEL-positive cells in the untreated group and the normal saline group. The numbers of RGCs in the untreated and normal saline groups were less than those in the normal control and sham-operated groups at 14 days or 28 days (P<0.05). Although the number of RGCs in the EGb 761 group was less than those in the normal control and sham-operated groups (P<0.05), it was more than those in the untreated and normal saline groups (P<0.05). N(95) amplitude in EGb 761 group was higher than those in the untreated and normal saline groups (P<0.05) and close to those in the normal control and sham-operated groups (P>0.05) at 14 days or 28 days. The number of RGCs was positive correlated to N(95) amplitude (r=0.859, P=0.001 5). CONCLUSION: EGb 761 can inhibit the apoptosis of RGCs in guinea pigs after optic nerve transection, thus protect the morphology and function of RGCs.

[Protective effect of Huangban Granule against light-induced retinal damage in rats].

OBJECTIVE: To observe the protective effect of Huangban Granule, a compound of traditional Chinese herbal medicine, on rats with retinal damage induced by light. METHODS: A total of 24 male Sprague-Dawley (SD) rats were randomly divided into normal control group, untreated group and Huangban Granule group. Retinal light damage was induced by exposure to constant white fluorescent light for 5 hours at an illumination of 2,800 Lux. The Huangban Granule was given 10 days before light exposure until the animals were sacrificed in Huangban Granule group, and an equal volume of distilled water for the rats in untreated group. Electroretinogram (ERG) was recorded in all animals 2 weeks after light exposure and the animals were sacrificed for histopathological examination of retina. The outer nuclear layers (ONLs) on the superior and inferior retina were counted. RESULTS: Fourteen days after light exposure, the ONLs on the superior retina were 3 to 6 in the untreated group and 7 to 9 in treatment group. There were 9 to 11 layers in normal group. The mean number of ONLs in the untreated group (4.68+/-1.64) was less than that in the treatment group (8.23+/-1.35) (P<0.01). B-wave amplitudes were (319.38+/-71.96) muV and (135.16+/-42.30) muV in Huangban Granule group and the untreated group respectively (P<0.01). A-wave amplitudes were (184.63+/-47.23) muV and (83.35+/-27.75) muV (P<0.01), and oscillatory potential amplitudes were (239.38+/-20.19) muV and (125.44+/-26.23) muV (P<0.01) respectively in the two group. There was no significant difference in implicit times of a-wave and b-wave among the three groups. CONCLUSION: Huangban Granule obviously protects both function and morphology of the retina from light-induced retinal damage in rats.

Edaravone (MCI-186), a free radical scavenger, attenuates retinal ischemia/reperfusion injury in rats.

AIM: To investigate the effect of edaravone (MCI-186), a free radical scavenger, against ischemia/reperfusion (I/R) injury in the rat retina. METHODS: Retinal ischemia was induced in male Sprague-Dawley rats by elevating intraocular pressure to 110 mmHg for 60 min. The rats were intraperitoneally injected with edaravone at a dose of 3 mg/kg at 30 min before ischemia, and then treated with edaravone (3 mg/kg, ip) twice daily for 1 or 5 d after I/R. The levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in the retinal tissues were determined on d 1 after I/R injury. The apoptosis of retinal neurons was detected on d 1 after I/R injury by terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling staining. The electroretinogram (ERG) was recorded on d 5 after reperfusion. RESULTS: Edaravone lowered MDA levels, raised SOD activity, and attenuated I/R-induced apoptosis of retinal neurons within the inner nuclear, ganglion cell, and outer nuclear layers of the rat retina. Moreover, edaravone suppressed I/R-induced reduction in a- and b-wave amplitudes of ERG. CONCLUSION: Edaravone can protect the retina from I/R injury in rats through reducing oxidative stress and inhibiting apoptosis of retinal neurons, which suggests that edaravone might be a potential choice for the treatment of I/R-induced eye disorders.

Characterization of a highly active promoter, PBbgpd, in Beauveria bassiana.

The promoter of glyceraldehyde-3-phosphate dehydrogenase (gpd) gene from Aspergillus nidulans (PgpdA) is widely used to direct expression of target genes constitutively in fungi. However, in some species, a heterogeneous promoter is found to be of low efficiency. To obtain a high-efficiency promoter for transformation of Beauveria bassiana, an entomopathogenic fungus widely used as an mycoinsecticide, a glyceraldehyde-3-phosphate dehydrogenase gene (Bbgpd) promoter, was cloned and characterized. Four deletion constructs (-2118, -1153, -726, and -354) of the 5'-upstream sequence of Bbgpd linked to a bar::gus fusion gene (phosphinothricin-resistance::beta-glucuronidase fused gene), which were used as selected marker gene and report gene, respectively, were generated. GUS activities of transgenic strains harboring -726, -1153, and -2118 deletion constructs were much stronger than that of the promoter of Aspergillus nidulans gpdA (PgpdA), with a twofold to threefold increase over that in the PgpdA construct. The -726 fragment was necessary to direct GUS expression in B. bassiana. No -354 transgenic progenies were obtained, possibly because it failed to initiate the transcription of bar::gus fusion gene. A remarkable increase of GUS activity was found between the -1153 and -726 constructs, indicating that some active transcriptional elements were located in this region. With a high expression level and relatively short sequence, PBbgpd can be used to drive target genes in B. bassiana transgenic research.