Objective To establish a high-throughput method that can simultaneously, quickly, and accurately detect main malaria-transmitting Anopheles species and their resistance genes in China, providing a high-throughput monitoring tool for monitoring the main malaria vectors in China after malaria elimination. Methods In different sampling locations, including Tengchong City, Yunnan Province; Wenchang City, Hainan Province; and Donggang City, Liaoning Province, adult specimens of mosquitoes, including Anopheles sinensis, Anopheles minimus, Anopheles dirus, and Anopheles anthropophagus, were collected. Polymerase chain reaction (PCR) technology and Sanger sequencing were employed to detect the ITS2, kdr (L1014), rdl (A296), and ace-1 (G119) genes in individual mosquitoes. For the analysis of mixed samples, an optimized multiplex PCR reaction system, custom-designed dual barcode primers, and next-generation sequencing (NGS) technology were utilized to detect the aforementioned genes. The consistency was assessed using Kappa consistency tests and Chi-square tests for multiple rates. Sensitivity, specificity, and the Youden index were calculated using a four-grid table calculation method. The costs associated with each step of the normal operational process for each method were statistically summarized, and the optimal quantity of mixed samples for detection was determined by a comprehensive approach. Results Conventional PCR amplification of gDNA from 300 mosquitoes resulted in 144 individuals of Anopheles sinensis, 53 individuals of Anopheles dirus, 62 individuals of Anopheles anthropophagus, and 41 individuals of Anopheles minimus, as identified by Sanger sequencing. The mutation frequencies of resistance genes kdr (L1014), rdl (A296), and ace-1 (G119) were found in 73, 27, and 41 specimens, respectively. Using a newly established multiplex PCR reaction system based on custom dual barcode and NGS sequencing technology, samples corresponding to Sanger sequencing were detected under different sample sizes. The two methods showed high consistency in the results (all Kappa>0.900). Multiple comparison tests showed significant differences in the consistencies of the two methods across different sample sizes N (40, 80, 160), N (120, 200, 240, 280), and N (300) (χ2=26.547, P<0.001). The new method demonstrated high sensitivity and specificity across various sample sizes, with the Youden index ranging from highest to lowest as follows: 1 (40, 80, 160)>0.994 (120)>0.990 (280)>0.988 (200)>0.987 (240)>0.985 (300). With an increase in sample size from 40 to 300, the cost per sequencing site for the new method decreased from 20.0 yuan to 8.3 yuan, while the cost per sequencing site for the conventional method decreased from 16.7 yuan to 15.4 yuan. The optimal mixed sample size for the new method was determined to be 280. Conclusion The newly developed multiplex PCR and barcode NGS detection method enables simultaneous screening of four major malaria vector mosquito species and the presence of mutations in the ace-1, kdr, and rdl resistance genes, exhibiting excellent stability, high sensitivity, and specificity. It allows for the efficient analysis of large sample sizes in a single run, offering a cost-effective alternative compared to other types of detection methods.

Objective To investigate the effect of microRNA(miR)-21 on proliferation and differentiation of murine pulmonary fibroblasts. Methods C57BL/6 mice of SPF grade (n=24) were randomly divided into Sham group and Pulmo?nary fibrosis model group with 12 mice in each group. Pulmonary fibrosis model was established by trans-tracheal jet ventila?tion of bleomycin into mice. The transcription levels of miR-21 were examined by quantitative real-time PCR in various pul?monary fibrosis tissues. Primary fibroblast were isolated and digested by Trypsin then inoculated into 6 well plate to reach confluence of 30%-50%. PBS (2.5μL), negative control stock solution and miR-21 mimic stock solution (20μmol/L) were added into Opti-MEM (50μL) as control group, blank group and miR-21 mimic group respectively.The cell viability was as?sessed by CCK-8. Expressions of ADAMTS-1 and TGF-β1 in the pulmonary fibroblasts were tested using Western blot. Re?sults The expression of miR-21 was significantly increased in lungs of mice in pulmonary fibrosis model group than that in sham group. Expression of miR-21 was higher in miR-21 mimic group than that in control group and blank group. Expres?sion of miR-21 was significantly higher with better cell viability in miR-21 mimic group than that in control group and blank group. The expression of ADAMTS-1 was significantly decreased in miR-21mimic group, while the expression of TGF-β1, a target gene of miR-21, was significantly increased in miR-21 mimic group compared with the other two groups. There is no significant different in expressions of ADAMTS-1 and TGF-β1 between control group and blank group. Conclusion Over?expression of miR-21 in pulmonary fibroblasts disrupts TGF-β1 signaling pathway by reducing expression of ADAMTS-1, which promotes the proliferation and differentiation of pulmonary fibroblast.

Objective To investigate methods and results of endovascular treatment in TASC (Ⅱ) D-type femoral artery occlusion. Methods From January 2012 to May 2013, 26 cases (26 branches) of superficial femoral artery occlusion with endovascular treatment of TASC (Ⅱ) D-type superficial femoral artery occlusion were retrospectively reviewed. The effi-cacy was evaluated through ABI, CTA, DSA and symptoms improved. Results 26 branches were treated with endovascular methods. Technical success rate was 80.7%(21/26), including 13 branche with stent implantation, 6 branches with Silver-hawk atherectomy and 2 branches with Viabahn stent implantation. All patients were followed up for a mean period of (10.3 ± 1.2)months, primary patency rates at 6 months were 69.2%in stent group, 66.7%in Silverhawk atherectomy group and 100%in Viabahn stent group. Conclusion Endovascular treatment of TASC (Ⅱ) D-type femoral artery occlusion can lead to satisfactory short term patency rates, and Viabahn stent is the latest treatment.

Objective To determine the prevalence of penicillinase-producing Neisseria gonorrhoeae(PPNG) and the distribution of blaTEM-135 gene variants in PPNG at several gonococcal antimicrobial surveillance sites in China, to compare N. gonorrhoeae multi-antigen sequence typing(NG-MAST)types of PPNG and its blaTEM-135 gene variants, and to assess the difference and association in NG-MAST types of blaTEM-135 gene variants among different regions. Methods A total of 572 N. gonorrhoeae isolates were collected at 6 gonococcal antimicrobial surveillance sites from Jiangsu, Shanghai, Zhejiang, Tianjin, Guangdong and Guangxi in 2012. After isolation, purification, and identification, cefalotin paper discs were used for detection of PPNG. DNA was extracted by QIAxtractor DX kits after cultivation of the PPNG strains. Then, mismatch amplification mutation assay (MAMA) PCR was performed to identify blaTEM-135 variants, and NG-MAST analysis to determine N. gonorrhoeae genotypes. Results Among the 572 N. gonorrhoeae strains, 38.1%(218/572) were identified as PPNG, and of the PPNG strains, 52.3% (114/218) were blaTEM-135 variants. The detection rate of PPNG at these surveillance sites from high to low was as follows: 51.7% (45/87, Zhejiang), 45.6%(36/79, Shanghai), 38.0% (78/205, Guangdong), 37.5% (12/32, Guangxi), 31.2% (24/77, Jiangsu) and 25.0%(23/92, Tianjin), and that of blaTEM-135 variants was as follows: 68.9%(31/45, Zhejiang), 58.3%(14/24, Jiangsu), 50.0%(39/78, Guangdong), 47.2%(17/36, Shanghai), 39.1%(9/23, Tianjin)and 33.3%(4/12, Guangxi). NG-MAST analysis showed that the ST2318, ST1768, ST1866, ST1053 and ST8726 types predominated among these bla TEM-135 variants, and a strong correlation was found between blaTEM-135 variants and some NG-MAST types, such as ST1768, ST1053 and ST8726 types. The distribution of NG-MAST types was significantly different between the surveillance site in Tianjin (in the Northern part of China) and the other sites (in the Southern part of China), but highly similar among the surveillance sites in Jiangsu, Zhejiang and Shanghai regions. Conclusions There is a high prevalence of PPNG and its blaTEM-135 variants at several gonococcal antimicrobial surveillance sites in China, with significant differences in NG-MAST genotype distribution of PPNG and its blaTEM-135 variants among different regions.

Objective To study the effect of application of plan-do-check-act (PDCA) on reducing the incidence of healthcare-associated infection(HAI) in department of neurosurgery.Methods Quality control circle activity group was established,programme of activities was formulated,four stages and ten steps of PDCA were adopted,incidences of H AI in department of neurosurgery before (September-November 2015) and after (May-July 2016) the implementation of PDCA were observed,causes were analyzed based on implementation of hand hygiene,aseptic technique manipulation,and environmental sanitation,countermeasures were found out,and continuous quality improvement was performed for 6 months.Results Comparison between before and after implementation of PDCA was conducted,incidence of HAI in department of nerosurgery decreased from 10.9％ to 5.8％,difference was significant(P＜0.05),control rate was 100％,incidence of HAI dropped by 46.8％;hand hygiene implementation rate increased from 27.2％ to 76.9％,aseptic technique implementation rate increased from 76.0％ to 96.9％,environmental sanitation increased from 51.0 ％ to 90.0 ％,differences before and after implementation were all statistically significant(all P＜0.001).Conclusion Quality control circle activities implemented jointly by multiple departments can reduce the incidence of HAI in department of neurosurgery,rules can be observed,measures can be further improved,it is worthy of clinical application.

OBJECTIVETo evaluate the mRNA and protein expressions of peroxiredoxin II (PrxII) in hepatocellular carcinoma (HCC) and their significance.

METHODSHCC was induced by aflatoxin B1 (AFB1) in 6 tree shrews (Tupaia belangeri chinensis). The expression levels of PrxII mRNA and protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot on HCC tissues and on their surrounding liver tissues (para-HCC). Biopsied liver tissues were taken before the HCC induction (pre-HCC) from the same animals and from a group of blank controlled animals that served as controls. Liver biopsy specimens from 18 cases of human HCC and from 17 healthy human volunteers were studied using the same methods.

RESULTSThe mRNA and protein expressions of PrxII in tree shrew HCC tissues were significantly higher than those in para-HCC and pre-HCC tissues, and also higher than those in the liver tissues from the control animals (all P < 0.05). The expression levels of PrxII mRNA and protein in human HCC tissues were also significantly higher than those in their para-HCC tissues and in the human normal liver tissues (P < 0.05).

CONCLUSIONPrxII might play an important role in hepatocarcinogenesis and might be used as a molecular target for HCC prevention and treatment.

Adult ; Aged ; Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Liver ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Liver Neoplasms, Experimental ; metabolism ; pathology ; Male ; Middle Aged ; Peroxiredoxins ; genetics ; Tupaiidae

OBJECTIVEA reversed superficial temporal artery auricular flap was presented to explore a new method for reconstructing the defects of the distal nose by microsurgical techniques.

METHODSA reversed superficial temporal artery auricular flap had been used on fifteen patients with nasal defects, including thirteen patients with alar defects and two patients with nasal tip defects. The reversed superficial temporal vessels of the flap were anastomosed with the recipient facial vessels. The size of the flap was 2.5 cm x 2. 0 cm - 4.0 cm x 2.5 cm, the length of the vascular pedicle was 5 - 8 cm, average 6.5 cm

RESULTSThe flap survived uneventfully in all fifteen patients.

RESULTSdemonstrated satisfactory symmetry between the reconstructed ala and the contralateral side as well as an excellent tip projection. The donor-site defect was minimal.

CONCLUSIONSThe reversed superficial temporal artery auricular flap offers an adequate length of vascular pedicle of the flap, it delivers a good solution to the problem of the vascular pedicle shortage of the proximal superficial artery auricular flap. This technique may become the top choice in the microvascular auricular transfer.

Adult ; Ear, External ; surgery ; Female ; Humans ; Male ; Middle Aged ; Nose ; surgery ; Nose Deformities, Acquired ; surgery ; Rhinoplasty ; methods ; Surgical Flaps ; blood supply ; Temporal Arteries ; surgery ; Tissue Transplantation ; Young Adult

Objective To compare the effect and safety of stenting versus directional atherectomy (DA) in the treatment of TASCⅡ A and B superficial femoral artery lesions.Methods 100 patients with TASC Ⅱ A and B lesions were divided into percutaneous transluminal stenting(PTS) group (n =50) and DA group (n =50).Patients were compared in terms of technical success rate,treatment success rate,first operation cost,postoperative ankle brachial index (ABI),limb salvage rate,survival,and patency.Results The technical success rate in both PTS and DA group was 100％.The treatment success rate was 98％ vs.86％,P＞0.05.Postoperative ABI:0.82 ±0.19 vs.0.80 ±0.27,P＞0.05.First operation cost:(34 820 ± 1 051) yuan vs.(45 635 ± 1 358) yuan,P ＜0.001;All patients were followed-up for up to 2-year,the cumulative patency rate was 81.6％ vs.72.9％ (P＞0.05).Limb salvage rate was 97.9％ vs.93.8 ％,P ＞ 0.05.Conclusion There were no significant differences in the effect and safety of PTS versus DA in the treatment of TASCⅡ A and B superficial femoral artery lesions.

OBJECTIVESTo study the biological function and its possible underlying mechanism of peroxiredoxin II (PrxII) in liver cancer cell line Hep3B.

METHODSTwo pairs of double-stranded small interfering RNA (siRNA) targeted on PrxII gene were transfected into Hep3B cells using LipofectamineTM 2000. After confirming the inhibited effects of these siRNAs through Quant SYBR Green polymerase chain reaction and Western blot, the biological characters of Hep3B cell were analyzed by flow cytometry analysis, MTT and colony formation assays. Furthermore, dichlorodihydrofluorescein diacetate (DCFH-DA) and thiobarbituric acid (TBA) assays, for measuring the products of oxidative reaction, such as the reactive oxygen species (ROS) and malondialdehyde (MDA), were applied to explore whether the antioxidant mechanism was involved in the effects of PrxII functioning on Hep3B cell.

RESULTSThe two pairs of siRNA significantly inhibited PrxII mRNA and protein expression. Compared to the mock and blank control groups, the two PrxII-silent groups showed decreased rates of cell growth and clone formation and increased rates of cell apoptosis. The numbers of the formed colonies were 42.0+/-2.8 and 40.5+/-0.7 respectively in the two PrxII-silent groups, while they were 121.5+/-2.1 and 130.0+/-1.4 in the mock and blank control groups (P less than 0.05). The levels of endogenous ROS and MDA were significantly higher in the two PrxII-silent groups than those in the mock and blank control groups (P less than 0.05).

CONCLUSIONPrxII might play an important role in the hepatocarcinogenesis, possibly through an antioxidant function which may provide a favorable microenvironment for cancer cell survival and progression.

Cell Line, Tumor ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Oxidative Stress ; Peroxiredoxins ; genetics ; RNA, Small Interfering ; Reactive Oxygen Species ; Signal Transduction ; Transfection

OBJECTIVETo investigate the effect of gene Af116609 on gastric cancer multi-drug resistance (MDR) by introducing it into gastric cancer multi-drug resistant (MDR) cell line SGC7901/VCR.

METHODSGene Af116609 was cloned from SGC7901/VCR by RT-PCR and its differential expression between gastric cancer MDR cells and its parental cells was displayed by Northern blot. The gene was introduced to gastric cancer cells by transfection of recombinant eukaryotic expression vector by electroporation. MTT assay in vitro was applied to investigate its effect on multi-drug resistance phenotype of gastric cancer cells.

RESULTSThe full length CDS of gene Af116609, as long as 327 bp, was cloned from gastric cancer MDR cell line SGC7901/VCR and its sequence was coincident with the hypothetical gene Af116609 in GenBank. It was overexpressed in MDR cells than its parental cells at mRNA level. In the MTT assay in vitro, the drug sensitive cells transfected with sense eukaryotic expression vector showed upregulated targeted gene, with increased resistance to vincristine, 5-fliorouracil and arabinoside, and decreased resistance to adriamycin, but no influence on resistance to methotrexate. However, the drug resistant cells transfected with anti-sense eukaryotic expression vector, showed down regulated targeted gene, with less resistance to all the five anticancer drugs to different degrees.

CONCLUSIONGene Af116609 is related to MDR phenotype of gastric cancer cells and may become a candidate molecular target to reverse the MDR of gastric cancer.

Antineoplastic Agents, Phytogenic ; pharmacology ; Autoantigens ; genetics ; Cell Line, Tumor ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Humans ; RNA, Small Cytoplasmic ; genetics ; Ribonucleoproteins ; genetics ; Stomach Neoplasms ; genetics ; pathology ; Vascular Endothelial Growth Factor A ; biosynthesis ; Vincristine ; pharmacology