Rapid assay using recombinase polymerase amplification and lateral flow dipstick for identifying Agrilus mali (Coleoptera: Buprestidae), a serious wood-boring beetle of the western Tianshan Mountains in China.

Agrilus mali stands as a significant wood-boring pest prevalent in Northeast Asia. Identifying this pest beetle is often hindered by insufficient efficient, rapid, on-site discrimination methods beyond examining adult morphological features. As a result, an urgent need arises for developing and implementing a rapid and accurate molecular technique to distinguish and manage the beetle. This study presents a straightforward, swift, highly specific, and sensitive method built upon recombinase polymerase amplification combined with a lateral flow dipstick (RPA-LFD). This method demonstrates the capability to promptly identify the beetle, even during its larval stage. RPA primers and probes were designed using the internal transcribed spacer 1 region. Through probe optimization, false-positive signals were successfully eliminated, with an accompanying discussion on the underlying causes of such signals. The RPA-LFD assays exhibited remarkable specificity and sensitivity, requiring as little as 10-3 ng of purified DNA. Furthermore, the extraction of crude DNA was achieved through immersion in sterile distilled water, thus streamlining the assay process. Achievable at temperatures ranging from 30 to 50 °C, the RPA-LFD assay can be executed manually without specialized equipment. By merging the RPA-LFD assay with DNA coarse extraction, A. mali can be detected within just 30 min. This current study effectively demonstrates the immense potential of RPA-LFD in quarantine and pest management. Additionally, it presents a universal technique for the rapid on-site diagnosis of insects, showcasing the wide applicability of this method.

The complete mitochondrial genome of Platygaster robiniae (Hymenoptera: Platygastridae): A novel tRNA secondary structure, gene rearrangements and phylogenetic implications.

Platygaster robiniae is economically important as a highly specific parasitoid of the invasive pest Obolodiplosis robiniae which was introduced into the Euro-Asia region in the last decade. Despite being a critical and specific parasitoid of the invasive pest O. robiniae and its use as an effective biocontrol agent, the absence of sequence information from P. robiniae have limited its genetic applications for pest management in forests. Mitochondrial (mt) genomes generally contain abundant nucleotide information and thus are helpful for understanding species history. Here, we sequenced the complete mt genome of P. robiniae using next generation sequencing, and annotated 13 protein-coding, 22 tRNA, and 2 rRNA genes and a 702 bp noncoding region. Comparative analysis indicated that this mt genome has a normal A + T content and codons use, however possessed both the expected and unique rearrangements. Ten tRNAs at four gene blocks COII-ATP8, COIII-ND3, ND3-ND5 and the A + T-rich region-ND2 were rearranged, including gene shuffles, transpositions and inversions. Notably, two genes tRNA Ser(UCN) and tRNA Leu(CUN) had undergone long-range inversions, which is the first record of this rearrangement type in the superfamily Platygastroidea. The D-loops of both tRNA Ile and tRNA Leu(CUN) were absent from the tRNA secondary structure, which has not been reported from hymenopteran previously. Phylogenetic analysis based with the maximum likelihood and Bayesian methods showed that P. robiniae grouped with other species of Platygastridae, and that the superfamily Platygastridea is sister to the other Proctotrupomorpha superfamilies. Our tree strongly supports the monophyly of the five superfamilies of Proctotrupomorpha. This study discovered some unique characters of P. robiniae, and contributes to our understanding of genome rearrangements in the order Hymenoptera.

[Correlation of B-type Natriuretic Peptide Level with Hemodynamic Parameters and Inflammatory Cytokines in Patients with Gram-negative Sepsis].

Objective To study the correlation of B-type natriuretic peptide(BNP)level with hemodynamic parameters and inflammatory cytokines in patients with Gram-negative sepsis,and further determine the main factors for the significant increase of BNP level. Methods The prospective study method was applied,and septic patients infected with Gram-negative bacteria from May 2017 to October 2019 were enrolled.The patients were divided into the BNP<2400 ng/L group and the BNP≥2400 ng/L group by taking the average value of BNP as the dividing point.The independent predictors of BNP≥2400 ng/L were analyzed by Logistic regression.Pearson correlation analysis was used to analyze the correlation between BNP and various indicators. Results A total of 106 patients with Gram-negative sepsis were included,among which 60 cases present with higher serum BNP levels than the average of(2398.45 ± 421.45)ng/L.Thus BNP≥2400 ng/L was considered as a significantly increased BNP level.Multiple logistic regression analysis showed that cardiac index(CI)[odds ratio (OR)=0.428,95% confidence interval (95%CI)=0.743-0.965,P=0.011],left ventricular ejection fraction(LVEF) (OR=0.394,95%CI=0.182-0.549,P=0.013),lactic acid (OR=1.983,95%CI=1.264-3.420,P=0.023),endotoxin (OR=6.146,95%CI=4.091-8.226,P=0.001),procalcitonin(PCT) (OR=6.513,95%CI=4.365-8.210,P=0.005) and cardiac troponin I(cTnI) (OR=1.144,95%CI=1.001-2.150,P=0.047) were independent predictors of BNP≥2400 ng/L in patients with Gram-negative sepsis.Pearson correlation analysis showed that BNP was negatively correlated with CI(R=-0.514,P<0.001)and LVEF (R=-0.552,P<0.001),whereas positively correlated with lactic acid (R=0.265,P=0.032),cTnI (R=0.204, P=0.036),PCT(R=0.801,P<0.001),and endotoxin(R=0.765,P<0.001). Conclusions In septic patients with Gram-negative bacterial infection,LVEF,CI,lactic acid,cTnI,endotoxin and PCT are all independent risk factors for the significant increase of BNP,and endotoxin and PCT were more significantly correlated with BNP increase.Endotoxin and inflammatory reaction may be the more important stimulators of BNP increase in septic patients with Gram-negative bacterial infection.

Effect of topical propranolol gel on plasma renin, angiotensin II and vascular endothelial growth factor in superficial infantile hemangiomas.

The effect of topical propranolol gel on the levels of plasma renin, angiotensin II (ATII) and vascular endothelial growth factor (VEGF) in superficial infantile hemangiomas (IHs) was investigated. Thirty-three consecutive children with superficial IHs were observed pre-treatment, 1 and 3 months after application of topical propranolol gel for the levels of plasma renin, ATII and VEGF in Department of General Surgery of Dongfang Hospital from February 2013 to February 2014. The plasma results of IHs were compared with those of 30 healthy infants of the same age from out-patient department. The clinical efficiency of topical propranolol gel at 1st, and 3rd month after application was 45%, and 82% respectively. The levels of plasma renin, ATII and VEGF in patients pre-treatment were higher than those in healthy infants (565.86 ± 49.66 vs. 18.19 ± 3.56, 3.20 ± 0.39 vs 0.30 ± 0.03, and 362.16 ± 27.29 vs. 85.63 ± 8.14, P < 0.05). The concentrations of VEGF and renin at 1st and 3rd month after treatment were decreased obviously as compared with those pre-treatment (271.51 ± 18.59 vs. 362.16 ± 27.29, and 405.18 ± 42.52 vs. 565.86 ± 49.66 P < 0.05; 240.80 ± 19.89 vs. 362.16 ± 27.29, and 325.90 ± 35.78 vs. 565.86 ± 49.66, P < 0.05, respectively), but the levels of plasma ATII declined slightly (2.96 ± 0.37 vs. 3.20 ± 0.39, and 2.47 ± 0.27 vs. 3.20 ± 0.39, P > 0.05). It was indicated that the increased renin, ATII and VEGF might play a role in the onset or development of IHs. Propranolol gel may suppress the proliferation of IHs by reducing VEGF.

[Effect of intermittent pneumatic compression on coagulation function and deep venous hemodynamics of lower limbs after rectal cancer resection].

OBJECTIVE: To investigate the effects of intermittent pneumatic compression (IPC) on coagulation function, deep venous hemodynamics and prevention of deep venous thrombosis (DVT) of lower limbs in patients after rectal cancer resection. METHODS: A total of 120 patients undergoing rectal cancer resection were randomly divided into non-IPC group (control group, n=60) and IPC group (n=60). The control group received routine treatment after resection and the IPC group received IPC based on the routine treatments. Prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), international normalized ratio (INR) and volume of D-dimer (D-D) were detected before operation and 1-, 3-, 5- and 7-day after operation. Meanwhile, blood flow velocity and caliber of external iliac vein, femoral vein and popliteal vein were examined by color Doppler ultrasound, then the average blood flow velocity and blood flow volume were calculated. RESULTS: Incidence of lower limb DVT was 13.3% (8/60) and 1.7% (1/60) in control group and IPC group respectively with significant difference (P<0.05). The differences in PT, APTT and INR were not significant (P>0.05) at 1-day after operation as compared to the preoperative level, while FIB and D-D both increased (P<0.05), all presented no significant difference among the two groups (P>0.05). PT shortened gradually (P<0.05), APTT and INR did not change significantly (P>0.05), FIB and D-D increased gradually (P<0.05), and no significant differences were found between the two groups at the same time point (all P>0.05). All the above parameters in the control group were significantly lower than those in IPC group (all P<0.05). CONCLUSIONS: IPC can improve hemodynamics indexes of deep veins of lower limb in patients after rectal cancer operation, and prevent the lower limb DVT. IPC is a safe, simple and convenient physical therapy.

[Effect of propranolol gel on infantile hemangiomas].

OBJECTIVE: To evaluate the efficacy and security of propranolol gel in treatment of Infantile hemangiomas. METHODS: 51 consecutive infants with hemangiomas from October 2010 to September 2011 in Department of General Surgery Fuzhou General Hospital of Nanjing Military Command were treated with propranolol hydrochloride 3% gel. Changes in hemangioma size, texture, color, tumor blood flow peak were recorded. RESULTS: The results were evaluated using Achauer system, responses of IHs to propranolol were considered scale I (poor) in 4 patient (17.24%), scale II (moderate) in 18 patients (24.14%), scale III (good) in 22 patients (44.83%) and scale IV (excellent) in 7 patients (13.79%). The response of superficial hemangiomas was significantly better than other hemangiomas (P < 0.05), and no significant differences in response among different primary sites (P > 0.05). CONCLUSIONS: Topical use of propranolol hydrochloride 3% gel is an effective option for superficial hemangiomas.

Increased expression of hLRH-1 in human gastric cancer and its implication in tumorigenesis.

Altered signaling pathways or deregulated transcription factors represent an important category of molecular events leading to aberrant gene regulation in gastric cancer, among which the role of WNT/beta-catenin pathway remains unclear. LRH-1 is a critical transcription factor in controlling cell proliferation via crosstalk with the beta-catenin signaling pathway. In order to gain a knowledge of the expression of hLRH-1v1 and hLRH-1 in gastric cancer, a Q-PCR analysis was carried out. Our results showed that in about 50 and 47.6% of 42 tested patients with gastric cancer, the mRNA expression of hLRH-1v1 and hLRH-1 was significantly upregulated, as compared with self-paired normal control, respectively. Besides, overexpression of hLRH-1 was shown to promote the proliferation of gastric adenocarcinoma cell SGC-7901 via induction of cyclin E1. Taken together, our present study demonstrated for the first time the increased expression of hLRH-1v1 and hLRH-1 in human gastric cancer, an alteration which may implicate in tumorigenesis.

[Hyperthermia enhanced the killing effect of 5-fluorocytosine on human colon cancer cell line transfected with cytosine deaminase gene].

OBJECTIVE: To investigate whether hyperthermia can enhance the killing effect of 5- fluorocytosine (5- FC) on human colorectal carcinoma cell lines SW480 transfected with carcinoembryonic antigen (CEA) tissue- specific cytosine deaminase (CD) gene in vitro,and study its mechanism. METHODS: Human colorectal carcinoma cell lines SW480 transfected with G1CEACDNa were cultured. The proliferated colonies were treated with the combined therapy of 5-FC and hyperthermia at a temperature of 43 degrees C for 30 min. After eight days, MTT was used to calculate the cellular survival rate,to analyze the killing effect of 5-FC combined with hyperthermia on SW480 cells transfected with CD gene. Flow cytometry was performed to analyze the cellular cycle and transmission electron microscope was used to observe the morphologic changes of SW480 cells after thermochemotherapy. RESULTS: Hyperthermia combined with 5-FC had an enhanced killing effect on SW480-CEACD cells than 5-FC alone (P< 0.05, t =2.403, n=9). Flow cytometry revealed that the proportion of S stage cell increased in the group treated with hyperthermia and 5- FC (P< 0.001, t =7.158, n=6). Transmission electron microscope showed apoptosis after thermo- chemotherapy. CONCLUSIONS: Hyperthermia can improve the anti- tumor effect of 5- FC on human colorectal carcinoma cell lines SW480 transfected with CD gene, and the cells were blocked at S stage of cellular cycle and apoptosis was induced following thermochemotherapy.