Toxic effects of permethrin on HMC3 microglia and its associated mechanism / 环境与职业医学

Background Permethrin is a commonly used pyrethroid insecticide and has been found to be potentially neurotoxic. Microglia are innate immune cells in the central nervous system and are involved in the development of a range of neurodegenerative diseases. Objective To observe possible toxic effects of permethrin on human microglia clone 3 (HMC3) in vitro and explore associated mechanism. Methods HMC3 were treated with 0, 10, 25, and 55 μmol·L−1 permethrin for 72 h. Cell cycle and apoptosis were measured using flow cytometry. Cyclin-dependent kinase 1 (CDK1), cyclin-dependent kinase inhibitor 1A (CDKN1A), cyclin B2 (CCNB2), cellular tumor antigen p53 (p53), factor-related apoptosis (FAS), caspase 3 (CASP3), and H2A histone family member X (H2AX) were detected by quantitative real-time PCR (qPCR). The differential genes and enrichment pathways of HMC3 after 0 and 25 μmol·L−1 permethrin treatment was analyzed by RNA sequencing. HMC3 was treated by 0, 10, 25, and 55 μmol· L−1 permethrin for 72 h. The content of nitric oxide (NO) in the supernatant was detected using Griess reagent. The secretion level of interleukin-6 (IL-6) was detected by enzyme linked immunosorbent assay (ELISA). The mRNA expression levels of mitogen-activated protein kinase (MAPK) pathway (including MAPK1, MAPK8, and MAPK14), interleukin-1β (IL-1β), IL-6, and matrix metalloproteinase (MMP) families (including MMP1, MMP2, MMP3, and MMP9) were detected by qPCR. The protein expressions of phosphorylated p38 mitogen-activated protein kinase (p-p38), phosphorylated extracellular signal-regulated kinase (p-ERK), IL-1β, IL-6, and MMP1 were detected by Western blot. Results HMC3 was arrested in G2/M phase after 0, 10, 25, and 55 μmol·L−1 permethrin treatment for 72 h, of which there was a statistically significant difference between the 55 μmol·L−1 permethrin treatment group and the control group (P<0.01), and the mRNA expression of CDKN1A was up-regulated according to the qPCR (P<0.05). There was no statistically significant difference in the proportions of apoptosis between the groups (P＞0.05). The RNA sequencing showed that the differential genes were enriched in the MAPK pathway, and the mRNA expressions of MAPK1, MAPK8, and MAPK14 were up-regulated after the permethrin treatment at 55 μmol·L−1 compared to the control group by qPCR (P<0.05). The Western blot revealed that, compared to the control group, the levels of p-p38 and p-ERK were increased after the 10 μmol·L−1 permetrin treatment (P<0.05), the p-ERK level was increased after the 25 μmol·L−1 permetrin treatment (P<0.05), and the p-p38 level was up-regulated after the 55 μmol·L−1 permetrin treatment (P<0.05). The secretion of NO in the supernatant of HMC3 increased after permetrin treatment compared to the control group (P<0.05), the mRNA and protein expressions and the secretion of IL-6 showed an upward trend, the mRNA and protein expressions of IL-1β were up-regulated (P<0.05), and the mRNA and protein expressions of MMP1 were up-regulated in the 25 and 55 μmol·L−1 permethrin groups (P<0.05). Conclusion Permethrin inhibits HMC3 cell proliferation in vitro, induces cell cycle arrest, activates MAPK pathway, and promotes the expression of inflammatory factors IL-1β and MMP1, which may be one of the mechanism of neurotoxicity induced by permethrin.

MiR-4443 promotes migration and invasion of breast cancer cells by inhibiting PEBP1 expression / 南方医科大学学报

OBJECTIVE@#To investigate the effect of miR-4443 expression on migration and invasion of breast cancer.@*METHODS@#We examined the expression of miR-4443 in breast carcinoma in situ and paired adjacent tissues from 3 breast cancer patients with high-throughput sequencing and verified the results using TCGA database. We also detected miR-4443 expressions using real-time quantitative PCR (RT-qPCR) in low invasive and highly invasive breast cancer cells (MCF-7 and MDA-MB-231 cells, respectively). The changes in apoptosis, migration and invasion of MCF-7 and MDA-MB-231 cells after transfection with miR-4443 mimics, mimics-NC, miR-4443 inhibitor or inhibitor-NC were analyzed using flow cytometry, wound healing assay and Transwell invasion assay. The target gene of miR-4443 was predicted by bioinformatics software and validated by a dual luciferase reporter gene system. RT-qPCR and Western blotting were performed to detect the expression of recombinant human phosphatidyl ethanolamine binding protein 1 (PEBP1) in the transfected cells.@*RESULTS@#The expression of miR-4443 was significantly higher in the breast cancer tissues than in the adjacent tissues (@*CONCLUSIONS@#MiR-4443 promotes the migration and invasion of breast cancer cells by inhibiting the expression of PEBP1, suggesting the possibility of suppressing miR-4443 expression as a potential therapeutic strategy for breast cancer.

Diagnostic value of combined detection of four-item urinary protein for early hypertensive kidney injury / 中国基层医药

Objective:To investigate the diagnostic value of combined detection of urine α 1-microglobulin(α 1-mG), N-acetyl-β-D-glucosaminidase (NAG), retinol binding protein (RBP) and β 2-microglobulin (β 2-mG) for early hypertensive kidney injury. Methods:From June 2016 to December 2018, 116 hypertension patients with renal damage (HRD group) and 44 cases with simple hypertension(HBP group) were selected in the Central People′s Hospital of Tengzhou in this study.And 36 cases of healthy people during the same period were selected as the control group.One hundred and sixteen cases of the HRD group were divided into Ⅰ-Ⅱ group (61 cases) and Ⅲ-Ⅴ group (55 cases) according to the classification of chronic kidney disease(CKD). The concentrations of α 1-mG, NAG, RBP and β 2-mG in urine were detected in patients and healthy people respectively.SPSS 19.0 software was used to perform statistical analysis. Results:The concentrations of urine α 1-mG, NAG, RBP, β 2-mG in the HRD group were (41.77±24.21)mg/L, (22.60±13.24)U/L, (2.86±1.73)mg/L, (1.76±0.95)mg/L, respectively, which in the HBP group were (12.49±8.10)mg/L, (13.45±8.61)U/L, (0.31±0.16)mg/L, (0.38±0.38)mg/L, respectively, which in the control group were (4.37±2.52)mg/L, (6.12±3.57)U/L, (0.29±0.17)mg/L, (0.28±0.15)mg/L, respectively.The concentrations of urine α 1-mG, NAG, RBP, β 2-mG in the HRD group were significantly higher than those in the HBP group( t=4.07, 4.25, 4.09, 4.03, all P<0.05) and the control group( t=3.15, 4.94, 2.49, 2.61, all P<0.05). The urine levels of α 1-mG, NAG, RBP, β 2-mG in phase Ⅰ-Ⅱ group were (21.62±13.45)mg/L, (21.96±12.49)U/L, (0.5±0.47)mg/L, (0.93±0.62)mg/L, respectively, which in the phase Ⅲ-Ⅴ group were (64.11±60.12)mg/L, (23.32±14.11)U/L, (5.48±4.77)mg/L, (2.68±2.55)mg/L, respectively.The concentrations of urine α 1-mG, NAG, RBP and β 2-mG in Ⅰ-Ⅱ group ( t=5.08, 4.99, 2.96, 1.66, all P<0.05) and Ⅲ-Ⅴ group ( t=3.95, 4.81, 4.33, 3.74, all P<0.05) were significantly higher than those in the control group.The levels of α 1-mG, RBP and β 2-mG in group Ⅲ-Ⅴ were higher than those in group Ⅰ-Ⅱ( t=5.37, 8.11, 4.52, all P<0.05). The positive detection rates of α 1-mG, NAG, RBP, β 2-mG and combination test in phase Ⅰ-Ⅱ group were 70.5%, 77.0%, 19.7%, 60.7%, 91.8%, respectively, which in the phase Ⅲ-Ⅴ group were 81.8%, 81.8%, 69.1%, 69.1% and 96.4%, respectively.The positive rate of urine α 1-mG, NAG, RBP and β 2-mG combination test was significantly higher than that of the single detection (phase Ⅰ-Ⅱ group: χ 2=7.71, 3.99, 61.4, 14.65; phase Ⅲ-Ⅴ group: χ 2=4.58, 4.58, 12.47, 12.47; all P<0.05). Conclusion:Urine α 1-mG, NAG, RBP and β 2-mG are important biochemical indicators in patients with early hypertensive kidney injury.The combined detection of the four tests has high diagnostic value in the diagnosis of early hypertensive nephropathy.

Clinical effects of perforating branch flaps of medial vastus muscle in repairing secondary wounds in donor sites of free anterolateral femoral perforator flaps / 中华烧伤杂志

Objective@#To investigate the clinical effects of perforating branch flaps of medial vastus muscle in repairing secondary wounds in donor sites of free anterolateral femoral perforator flaps.@*Methods@#From August 2014 to December 2016, 12 patients (8 males and 4 females, aged 35-72 years) with skin and soft tissue defects of extremities associated with tendon and bone exposure were treated in Hanzhong Central Hospital. The sizes of the primary wounds after debridement were 10 cm×8 cm-22 cm×14 cm, and the wounds were repaired with 12 cm×10 cm-24 cm×16 cm free anterolateral femoral perforator flaps. The anterolateral femoral donor sites, which were 8.0 cm×4.0 cm-14.0 cm×7.5 cm in the secondary wounds after skin extensional suture, were repaired with perforating branch flaps of medial vastus muscle in the size of 9.0 cm×5.0 cm-15.0 cm×8.5 cm. The medial femoral donor sites were sutured directly.@*Results@#All the perforating branch flaps of medial vastus muscle and free anterolateral femoral perforator flaps survived in 12 patients. Following up for 6 to 12 months, the medial femoral perforator flaps had good local shape and texture. The flaps of 8 patients without cutaneous nerve transection were sensitive. The sensation of the flaps of the other 4 patients gradually recovered, and the functions of the ipsilateral knee joints were normal.@*Conclusions@#The medial femoral perforator flap has a stable anatomy and abundant blood supply, which can be used to repair the secondary wound in the donor site of the free anterolateral femoral perforator flap conveniently. It is safe and easy to be popularized. Moreover, it has a good shape and function after operation.

Clinical application effect of blood circulation enhancement technique in repairing large area of skin and soft tissue defects of extremities with super large free anterolateral thigh flap / 中华烧伤杂志

Objective@#To observe the clinical application effect of blood circulation enhancement technique in repairing large area of skin and soft tissue defects of extremities with super large free anterolateral thigh flap.@*Methods@#From March 2014 to March 2017, 6 patients with large area of skin and soft tissue defects of extremities were hospitalized in our unit, including 5 males and 1 female, aged 27-65 years, 1 case of electric injury, 2 cases of coal burn, 3 cases of traffic injury, 2 cases involving upper limb, and 4 cases involving lower limb. After debridement, the wound area ranged from 26 cm×8 cm to 36 cm×15 cm, and the bone exposure area ranged from 24 cm×7 cm to 35 cm×14 cm. The blood circulation enhancement technique was used when the wound with bone exposure was repaired with super large free anterolateral thigh flap. The area of flaps ranged from 28 cm×10 cm to 38 cm×16 cm. The donor site of flap and the primary wound without bone exposure were repaired with medial thigh split-thickness skin graft of the donor leg of flap. The blood circulation enhancement technique mode during operation and the survival of flaps after operation were recorded, and the recovery of donor and recipient areas and the occurrence of complications were followed up.@*Results@#Three patients were treated with simple vascular supercharging technique during flap transplantation, and the other 3 patients were treated with vascular supercharging and turbocharging technique during flap transplantation. All the flaps survived well in 6 patients without vascular crisis. Follow-up for 3 to 12 months after surgery showed that the blood flow of the flaps was good and the depth and superficial sensation recovered to varying degrees. Except for 1 case of upper limb flap, the other flaps had no obvious swelling and needed no second thinning. There were only depressed scars in the donor sites, and no obvious scar hyperplasia in the area without bone exposure repaired by the skin grafts. No short-term or long-term complications were found.@*Conclusions@#The application of blood circulation enhancement technique in repairing large area of skin and soft tissue defects of extremities with super large free anterolateral thigh flaps provides reliable blood supply for the flaps and results in good effect after operation, which is worth popularizing.

Ultrashort Echo Time MRI (UTE-MRI) Quantifications of Cortical Bone Varied Significantly at Body Temperature Compared with Room Temperature

PURPOSE: To investigate the temperature-based differences of cortical bone ultrashort echo time MRI (UTE-MRI) biomarkers between body and room temperatures. Investigations of ex vivo UTE-MRI techniques were performed mostly at room temperature however, it is noted that the MRI properties of cortical bone may differ in vivo due to the higher temperature which exists as a condition in the live body. MATERIALS AND METHODS: Cortical bone specimens from fourteen donors (63 ± 21 years old, 6 females and 8 males) were scanned on a 3T clinical scanner at body and room temperatures to perform T1, T2*, inversion recovery UTE (IR-UTE) T2* measurements, and two-pool magnetization transfer (MT) modeling. RESULTS: Single-component T2*, IR-T2*, short and long component T2*s from bi-component analysis, and T1 showed significantly higher values while the noted macromolecular fraction (MMF) from MT modeling showed significantly lower values at body temperature, as compared with room temperature. However, it is noted that the short component fraction (Frac1) showed higher values at body temperature. CONCLUSION: This study highlights the need for careful consideration of the temperature effects on MRI measurements, before extending a conclusion from ex vivo studies on cortical bone specimens to clinical in vivo studies. It is noted that the increased relaxation times at higher temperature was most likely due to an increased molecular motion. The T1 increase for the studied human bone specimens was noted as being significantly higher than the previously reported values for bovine cortical bone. The prevailing discipline notes that the increased relaxation times of the bound water likely resulted in a lower signal loss during data acquisition, which led to the incidence of a higher Frac1 at body temperature.

Clinical value of serum AFP-L3 and GP73 in the diagnosis and assessment of primary liver cancer / 中国基层医药

Objective To investigate the clinical value of serum alpha fetoprotein -L3 ( AFP-L3) and Golgi body protein (GP73) in the diagnosis and assessment of primary liver cancer.Methods From October 2013 to May 2017,125 cases with primary liver cancer in Tengzhou Central People's Hospital were selected as hepatocellular carcinoma(HCC) group,and 60 patients with benign liver lesions were selected as benign lesion group ,in the same period,60 healthy people were selected as control group.The separation of AFP -L3 was through microspincolum method,enzyme-linked immunosorbent assay was used to detect GP 73.Results The serum AFP -L3 and GP73 levels in the HCC group were (33.16 ±9.17)%,(309.61 ±10.16) μg/L,respectively,which in the benign lesion group were (4.19 ±1.97)%,(43.18 ±9.17) μg/L,respectively,which in the control group were (3.25 ± 1.26)%,(38.52 ±7.09)μg/L,respectively.The serum levels of AFP-L3 and GP73 in the HCC group were signif-icantly higher than those in benign lesion group and control group (t ＝13.264,9.065,15.328,10.826,all P＜0.05).The serum levels of AFP-L3 and GP73 between the benign lesion group and control group had no statistically significant differences (t＝0.635,0.772,all P＞0.05).The sensitivity,specificity and accuracy of AFP -L3 in the diagnosis of primary liver cancer were 74.2%,87.5%and 91.2%,respectively.The sensitivity,specificity and accu-racy of GP73 in the diagnosis of primary liver cancer were 84.7%,81.4%and 92.4%,respectively.The sensitivity, specificity and accuracy of combined detection of them in diagnosis of primary liver cancer were 98.2%,89.6%and 97.1%,respectively.The sensitivity and accuracy of the combined tests were significantly higher than AFP -L3 or GP73 alone (t＝5.026,4.114,3.018,2.776,all P＜0.05).According to TNM staging,125 patients with HCC were divided into stage Ⅰ+Ⅱ and Ⅲ+Ⅳ.The serum AFP -L3 and GP73 levels in patients with stage Ⅰ+Ⅱ were (28.25 ±8.26)%,(202.86 ±9.24) μg/L,respectively,which were significantly lower than those in patients with stage Ⅲ+Ⅳ[(43.17 ±10.24)%,(398.62 ±11.35)μg/L] (t ＝3.772,3.182,all P ＜0.05).Conclusion The serum levels of AFP -L3 and GP73 in patients with primary liver cancer are obviously higher.Combined detection can improve the sensitivity and accuracy of diagnosis ,and the degree of increase is related to the severity of the patients.

Effects of axial vascular network flap of scalp or anterolateral thigh perforator flap with fascia lata on repairing defects after radical resection of scalp carcinoma in patients / 中华烧伤杂志

Objective@#To observe the effects of axial vascular network flap of scalp or anterolateral thigh perforator flap with fascia lata on repairing defects after radical resection of scalp carcinoma in patients.@*Methods@#From February 2006 to December 2015, twenty-one patients with scalp carcinoma were admitted to our hospital, and the carcinoma invaded external lamina or full-thickness of skull and dura mater. After perfect preoperative examination, carcinoma and scalp tissue in 3 to 5 cm from the edge of carcinoma, external lamina or full-thickness of skull and invaded dura mater were resected and sentinel lymph nodes around carcinoma were cleaned in 3 to 4 days after admission. The postoperative defects with size reached from 11 cm×8 cm to 22 cm×18 cm. The flap transplantation was performed at the same time when quick frozen pathological examination results of resected scalp carcinoma margin tissue, skull, dura mater margin and basal tissue, and sentinel lymph nodes showed completely negative. Defects in 3 elderly patients were repaired by single or multiple axial scalp vascular network flaps, with the resected flaps size ranged from 12 cm×7 cm to 19 cm×14 cm. Defects in the other 18 patients were repaired by anterolateral thigh perforator flaps with fascia lata, with the resected flaps size ranged from 13 cm×10 cm to 23 cm×19 cm and the resected fascia lata size ranged from 8 cm×7 cm to 10 cm×10 cm. The head donor site of flap was repaired by medium thickness skin of head and back; the thigh donor site of flap was repaired by medium thickness skin of thigh on the same side. All patients gave up postoperative radiotherapy, chemotherapy, and other follow-up treatments.@*Results@#After operation, the flap and skin in all patients survived completely, with no vascular crisis or other condition. During the follow-up for 6 months to 9 years, all patients showed good appearance except for baldness in operation area of head, with no obvious malformation in head donor site of flap and skin, no swollen external hernia in the brain tissue, and no local recurrence or distant metastasis of carcinoma. The appearance of thigh donor site of flap and skin was good, with normal muscle strength and movement of lower limbs.@*Conclusions@#Patients with scalp carcinoma were performed with radical resection of carcinoma, and axial vascular network flap of scalp or anterolateral thigh perforator flap with fascia lata were applied to repair the postoperative defects, with good appearance of head operation area and no local recurrence or distant metastasis of carcinoma.

Isolation and characterization of polymorphic microsatellite markers of Oncomelania hupensis / 中国血吸虫病防治杂志

Objective To isolate the microsatellite DNA sequences of Oncomelania hupensis and analyze the polymorphic microsatellite loci.Methods The digested genomic fragments were hybridized with biotinylated oligonucleotide probes.The target fragments moleculars were captured and enriched.Then these fragments were cloned and sequenced.The suitable microsatellite loci were chosen and the polymorphism was screened by PAGE gel electrophoresis.Results A total of 205 microsateilite DNA sequences were obtained (GenBank accession numbers :GU204044～GU204248).The percentage of perfect microsatellite DNA sequence was 36.10% (74/205),with imperfect sequence as 49.76% (102/205) and compound sequence as 14.15% (29/205).Twenty typical microsatellite sequences were selected to design amplifying primers,and 13 microsatellite loci were found to be polymorphism.Conclusion A total of 205 microsatellite DNA sequences of Oncomelania hupensis are isolated and first reported,which will be useful for population genetic and mapping studies of Oncomelania hupensis.

Fumigating insecticidal activity of 5 essential oils against Culex pipiens quinquefasciatus / 第二军医大学学报

Objective:To observe the fumigating insecticidal activity of 5 essential oils(asteraceae oil,rutaceae oil,mentha piperita oil,carvacryl oil and citronella oil) against Culex pipiens quinquefasciatus.Methods: Fumigating insecticidal activity was investigated by airtight fumigation in conical flask.Results: After different fumigating time,5 essential oils showed different toxity against Culex pipien quinqucfasciatus.Of 5 essential oils,rutaceae oil was the most toxic, with the LC 50 values being 0.013(0.5 h),0.055(4 h) and 0.058(24 h); asteraceae oil was the lowest toxic one, with the LC 50 values being 0.948(0.5 h),0.427(4 h) and 1.711(24 h).When LC 95 values of the 5 essential oils were used to treat Culex pipiens quinquefasciatus,the shortest fumigating time appeared in carvacryl oil(6.087 min) and the longest in citrinella oil(21.143 min).Conclusion: All the 5 essential oils have considerable insecticidal effects against Culex pipiens quinquefasciatus.Rutaceae oil and carvacryl oil are better than the others,which provides basic informations for the related field experiments.

Genetic Variation of Anopheles dirus A and D (Diptera:Culicidae) in China: Inferred by mtDNA-COⅠ Gene Sequences / 中国寄生虫学与寄生虫病杂志

Objective To interpret genetic variation and population structure of Anopheles dirus A and D from China by molecular marker. Methods Samples included An. dirus A of Hainan laboratory colony (n=13), and field specimen from Mengla (n=17) and Jiangcheng (n=17) in Yunnan Province. The specimens were identified by PCR assay before study. mtDNA-COⅠ region was amplified and sequenced. Genetic variation and population structure was estimated according to sequence data. Results The mtDNA-COⅠ gene with a length of 959 bp was analyzed. There were three haplotypes in An. dirus A and six haplotypes in An. dirus D. The above haplotypes distributed in three populations unif-ormly. The average number of pairwise differences within Mengla population (7.441 2) was greater than that of Jiangcheng (1.279 4) and Hainan (1.051 3) populations, which suggested that the level of genetic divergence was the highest within Mengla population. The result of hierarchical AMOVA estimation showed a limited geneflow (Fst=0.799 9), therefore the variation level in a population (20.01%) was smaller than among the populations (79.99%). Conclusion The inter-specific genetic variation between An. dirus A and D in China was small and the level of divergence among individuals was high.

Molecular Identification of Anopheles maculatus Complex from China / 中国寄生虫学与寄生虫病杂志

Objective To establish the molecular identification of five members in Anopheles maculatus complex from China. Methods Different rDNA-ITS2 regions of An. maculatus complex were sequenced and analyzed. The species specific primers were designed, and PCR assay was used for the identification. Results The length and GC contents of ITS2 were 328 bp, 58.54% in An. pseudowillmori, 330 bp, 57.85% in An. maculatus, 337 bp, 59.05% in An. willmori, 334 bp, 58.68% in An. dravidicus, and 338 bp, 57.69% in An. sawadwongporni, respectively. The intra-species ITS2 sequences were conservative. The ranges of divergence level among five members were from 9.7% to 18.9% . Five distinct specific fragments were amplified by PCR assay using five species specific primers and 5. 8S primer. The length was 119, 186, 231, 327 and 406 bp respectively. Conclusion The diagnostic PCR assay based on ITS2 divergence to distinguish five members of An. maculatus complex was simple and reliable.

ON MOLECULAR IDENTIFICATION AND TAXONOMIC STATUS OF ANOPHELES LESTERI AND ANOPHELES ANTHROPOPHAGUS IN CHINA (DIPTERA:CULICIDAE) / 中国寄生虫学与寄生虫病杂志

Objective] To clarify the taxonomic status of Anopheles lesteri and An.anthropophagus in China. [Methods] Using molecular identification (PCR assay and rDNA\|ITS2 sequencing) to examine the field anopheline mosquito specimens from Liaoning and Shandong. According to the ITS2 sequences, molecular phylogenetic tree was made. [Results] According to the molecular identification, An.lesteri and An.anthropophagus were distributed both in Liaoning Province and Shandong Province. The length and GC content of rDNA\|ITS2 sequence were 451 bp, 46 2% in An.lesteri (n=6), and 448 bp, 46 0% in An.anthropophagus (n=10), respectively. The ITS2 sequences from presentation sites were same in An.lesteri, while the intraspecies difference in An.anthropophagus was 0 88%. The specific difference between An.lesteri and An.anthropophagus was 25 7%. By analyzing molecular phylogenetic tree, the relationship between An.lesteri and An.sinensis, An.anthropophagus and An.liangshanensis was found to be closer. [Conclusion] According to the molecular identification, it was defined that An.lesteri and An.anthropophagus were sympatric independent species in China.

Isolation of Microsatellite DNA and the Polymorphic Locus Screening from Phlebotomus chinensis (Diptera:Psychodidae) / 中国寄生虫学与寄生虫病杂志

Objective To isolate the microsatellite DNA of Phlebotomus chinensis and screen the polymorphic loci.Methods Genomic DNA fragments were hybridized with biotinylated oligonucleotide probes(AAT)17,(GA)25,(CCT)17,and(TG)18.The hybridized fragments were captured with Vectrex Avidin D and enriched by centrifugal ultrafiltration with ultra-4-column ultrafiltrate.The target fragments were amplified,cloned and sequenced.The suitable microsatellite loci were chosen in Ph.chinensis's library to establish PCR amplification assay.The polymorphism screening was conducted by PAGE with Ph.chinensis field specimens.Results The enrichment protocol used in this study was efficient,with a percentage of recombinant clone as 78.6%.There were 118 microsatellite sequences in library,the GenBank accession numbers were from FJ919812 to FJ919932(except GenBank accession numbers FJ919833,FJ919836,and FJ919869).There were 72 typical microsatellite sequences occupying 61.0% and the rest were 46 nontypical microsatellite sequences in the library.Twenty-two loci were chosen to polymorphism screening and PAGE showed that 14 loci were polymorphic.The loci of dinucleotide repeat were more polymorphic than those of trinucleotide and polynucleotide repeat.Conclusion The microsatellite-containing library of Ph.chinensis has been constructed with 118 sequences,and 14 new polymorphic microsatellite loci are reported.