Objective To investigate the role of ribosomal protein S7(rpS7) in the defense of Anopheles dirus against infection. Methods rpS7 was amplified from Anopheles dirus hemocytes with degenerated primers designed according to the conservative region of S7, rpS7 was then cloned using T/A cloning kit and the inserted fragment was sequenced. The difference of the transcript abundance of rpS7 from Anopheles dirus hemocyte among non-blood-fed (N), normal-blood-fed (B) and Plasmodium yoelii infected groups (I) was also analyzed by RT-PCR and gel scanning system at d1, d2, d3, d4, d7 and d11 after blood feeding. Results There is no significant difference of rpS7 signal between the three groups. Conclusion Anopheles dirus S7 can be used as an internal control to study the role of Anopheles dirus related immune factors in Plasmodium infection.

Objective To explore the contamination of sewage from hotels and restaurants by Salmonella typhi. Methods 327 sewage samples collected from hotels and restaurants and 176 sewage samples collected from some residential area were detected for Salmonella typhi based on the national standard methods. Results The positive rate of Salmonella typhi in the sewage samples from hotels and restaurants (3.67%) was significantly higher than that from residential area(0.57%), P=0.04(exact probability test).12 strains of Salmonella typhi belonging to 4 sero-groups and 9 serotypes were found in 12 sewage samples collected from hotels and restaurants.One strain of Salmonella typhimurium was only found in one sewage sample collected from residential area. The positive rate of Salmonella typhi in the sewage samples collected during January-March (5.47%) was significantly higher than that during April-June(0.79%) among total 327 sewage samples collected from hotels and restaurants, P=0.033(exact probability test). Conclusion The contamination of sewage from hotels and restaurants by Salmonella typhi and its potential risk to human health should be paid more attention to.

Objective To study the role of cytidine-phosphate-guanosine oligodeoxynucleotide(CpG ODN) on the development of Plasmodium liver stage.Methods Plasmodium yoelii BY265 18S rRNA was cloned,and the TaqMan real-time PCR was established on P.yoelii BY265 18S rRNA and mouse GAPDH as quantitative analysis model.The model was tested by the level of liver Plasmodium load with the liver cDNA in BALB/c mice infected by salivary gland sporozoites for 42 hours.Twelve BALB/c mice were randomly divided into CpG group,CpG control group and PBS control group which were injected respectively by ODN1826 30 ?g,ODN1826 control 30 ?g and 0.01 mol/L PBS 200 ?l via vena caudalis.Twenty-four hours later,each mouse was inoculated with 100 sporozoites.Mice were sacrificed in 42 hours after infection,and the liver load of Plasmodium was analyzed by TaqMan real-time PCR.Results The cloned Py BY265 18S rRNA gene showed 98% similarity to Py 17XNL.The quantitative analysis model consisted by 18S rRNA and GAPDH showed positive correlation between the level of liver Plasmodium load and the sporozoite inoculation dose to mice.The Plasmodium load in CpG ODN pre-treated mice was reduced to one fifth of the control group(0.28/1.33)(P

Objective　To explore the relationship between hemolymph phenol oxidase and the melanization of Plasmodium yoelii oocysts in Anopheles dirus. Methods　An Anopheles dirus-Plasmodium yoelii system was used Anopheles dirus were divided into 3 groups, that is, non-blood-fedding (N), normal-blood-fedding (B) and infected-blood-fedding (I). The activities of MPO and o-DPO in hemolymph from 3 groups were determined with native polyacrylamide gel electrophoresis (PAGE) and density scanning at 5, 7, 11 and 15 d after blood feeding. Results　Both MPO and o-DPO activity were significantly higher in group I than group N and B (P<0.05). But with the melanization of Plasmodium yoelii oocysts, both MPO and o-DPO activity in group I were decreased in comparison with group N, especially on the 15 th day after infected-blood feeding. MPO and o-DPO activity in group B were significantly stronger than those of group N. Conclusion　Blood feeding and infection of Plasmodium yoelii both can activate the cascade. The heamolymph phenol oxidase may play an important role in the melanization of Plasmodium yoelii oocysts in Anopheles dirus.

Objective To analyze the relationship between the TEP1 gene of Anopheles stephensi and melanotic encapsulation of Plasmodium yoelii induced by anti-malaria drug nitroquine. Methods Haemolymph samples from three groups of An. stephensi fed with sucrose solution, Plasmodium-infected blood and nitroquine, respectively, were collected at the 1st, 2nd, 3rd and 4th day after drug adminstration. Degenerate primers were designed according to the conserved amino acid sequence within TEPs of the mosquitoes. Fluorescent quantitation PCR was used to detect the variation of TEP1 gene transcript induced by nitroquine. The melanization of oocysts was observed by light microcopy. Results TEP1 gene was cloned, the predicted amino acid residues harbored a highly conserved canonical thioester motif GCGEQ. The fluorescent quantitation PCR revealed that nitroquine induced an up-regulation of TEP1 activity. The transcription of TEP1 gene in nitroquine treated group (2.423) was significantly higher than that of the infected blood-fed group(1.036) at the 3rd day after nitroquine treatment (P

Objective To observe the effect of blood feeding and plasmodium yoelii infection on the transcript abundance of ribosomal protein S7 from Anopheles dirus hemocyte Methods Anopheles dirus of 3～5 days old were divided into normal group (N), blood feeding group(B) and Plasmodium yoelii infection group(I) Hemocytes of 50 Anopheles dirus from each group were collected by centrifuge method on days 1, 2, 3, 4, 7 and 11 after blood feeding, respectively Then, all hemocyte samples were used for total RNA isolation and analyzed by RT PCR Finally, the same volume(10 ?l) of all the PCR products from each group was used for agarose gel electrophoresis and the data obtained were analyzed statistically Results There was no significant difference in ribosomal protein S7 signal between the three groups Conclusion Similar to Anopheles gambiae and human rpS7, Anopheles dirus rpS7 might be also used as an internal control for the studies of the role of Anopheles dirus related immune factors in attacking Plasmodium infection

Objective To construct cDNA library from adults of Anopheles dirus for cloning the immune genes or related genes for malaria parasites development. Methods The mRNA of adult Anopheles dirus was isolated. The library was constructed by using the Zap Express vector(Stratagene) and the quality was evaluated. Results The efficiency of the library was 2.1?10 6 pfu/ml with 98% clones positive. The average length of the insert fragment was over 1 kb. Conclusion cDNA library of adult Anopheles dirus with high efficiency can be constructed by using the Zap Express library construction Kit (Stratagene).

Objective To investigate the expression patterns of serine proteases Adsp1 and Adsp3 in the main tissue of Anopheles dirus and the effects of blood feeding and Plasmodium infection on the transcript abundance of Adsp1 and Adsp3 from Anopheles dirus haemocytes. Methods Haemocytes were collected by centrifugation. The midguts and salivary glands were dissected from 50 adult female Anopheles dirus of 3-5 d old for the extraction of total RNA for amplification by RT PCR. Anopheles dirus of 3-5 d old were divided into normal group (N), blood feeding group (B), and Plasmodium yoelii infection group (I). Haemocytes of 50 adult female Anopheles dirus from each group after blood feeding were also collected, and the total RNA was isolated at 1, 2, 3, 4, 7, and 11 d, respectively. Then, the same volume (10 ?l) of total RNA was analyzed by semi quantitative RT PCR with Ads7 as the internal control. Agarose gel analysis was performed on PCR products from each group. Results Expression of both Adsp1 and Adsp3 mRNA was found in the haemocytes and salivary glands, but not in the midguts. Semi quantitative RT PCR indicated that transcript abundance of AdsP1 and AdsP3 from Anopheles dirus haemocytes was enhanced after blood feeding and Plasmodium yoelii infection, especially during melanotic encapsulation of Plasmodium yoelii . Conclusion AdsP1 and AdsP3 may be involved in the melanotic encapsulation response of Plasmodium yoelii by Anopheles dirus , and may be the prophenoloxidase activating enzyme of Anopheles dirus .

Objective To investigate the clinicopathologic features of gastrointestinal neuroendocrine carcinomas. MethodsThe diagnosis and treatment results of 45 cases were studied, and clinicopathologic features and immunohistochemical expressions of NSE, Syn and CgA were detected.ResultsMicroscopically carcinomas were divided into three types: type Ⅰ(25 cases), type Ⅱ(10 cases) and type Ⅲ(10 cases). The histologic categories were correlated with lymph node metastasis significantly( P0.05). The 5-year survival rate for type Ⅰ, type Ⅱ and type Ⅲ was 70%, 65% and 52%, respectively.ConclusionsThe combination of NSE, Syn and CgA immunohistochemical stainnig is necessary for the diagnosis of gastrointestinal neuroendocrine carcinomas. The histologic classification is coincident with the requirement of clinical treatment and prognosis.

Objective To provide more objective reference in diagnosis and prognosis of coronary heart disease(CHD)combined with heart failure(HF)by analyzing features of pulse graph parameters improved with moving average line method of patients with CHD combined with HF.Methods From September 2018 to December 2020,a random sampling method was used to collect 78 inpatients with CHD and 73 inpatients with CHD combined with HF in department of cardiology of Shu Guang Hospital Attached to Shanghai TCM University,Shanghai Municipal Hospital of Traditional Chinese Medicine and Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine as well as 78 healthy people from physical examination center of Shu Guang Hospital Attached to Shanghai TCM University and Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine.Smart TCM-I type pulse digital acquisition analyzer was used to collect pulse samples.χ2 test,LSD test,Dunnett's T3 test and Kruskal Wallis test were used to analyze the differences of pulse graph parameters among the groups improved with moving average line method.Results ①Results of pulse graph parameters about chronaxy:Compared with those in healthy people,t5 of patients with CHD decreased significantly(P<0.01),t,t3,t5 of patients with CHD combined with HF increased significantly(P<0.01).Compared with those in patients with CHD,t5 of patients with CHD combined with HF increased significantly(P<0.01),t3 increased(P<0.05).②Results of pulse graph parameters about ratio of chronaxy:Compared with those in healthy people,t1/t,w/t,tf/4/t,tf/3/t4,tf/4/t4,tf/5/t4,tf/3/tmax,tf/4/tmax,tf/5/tmax,tf/6/tmax of patients with CHD decreased significantly(P<0.01),tmax/t,w/t4,tf/3/t,tf/5/t,tf/6/t,tf/6/t4 decreased(P<0.05),t0.8/tmax increased(P<0.05),w/t,t0.9/t,t0.8/t,tf/3/t,tf/4/t,tf/5/t,tf/6/t,tf/3/t4,tf/4/t4,tf/3/tmax,tf/4/tmax,tf/5/tmax,tf/6/tmax of patients with CHD combined with HF decreased significantly(P<0.01),t1/t,tmax/t,w/t4,t0.9/t4 decreased(P<0.05),t3/t4 increased significantly(P<0.01).Compared with those in patients with CHD,t0.9/tmax of patients with CHD combined with HF decreased significantly(P<0.01),t0.9/t,t0.8/tmax decreased(P<0.05),t3/t4 increased significantly(P<0.01).③Results of pulse graph parameters about amplitude:Compared with those in healthy people,h4 of patients with CHD decreased(P<0.05),h3,h4 of patients with CHD combined with HF decreased significantly(P<0.01).Compared with those in patients with CHD,h5 of patients with CHD combined with HF decreased significantly(P<0.01),h3 decreased(P<0.05).④Results of pulse graph parameters about ratio of amplitude:Compared with those in healthy people,h4/h1 of patients with CHD decreased(P<0.05),h3/h1,h4/h1 of patients with CHD combined with HF decreased significantly(P<0.01).Compared with those in patients with CHD,h5/h1 of patients with CHD combined with HF decreased significantly(P<0.01).Conclusion Patients with CHD combined with HF characterize a weakening arterial compliance and an increasing peripheral resistance.Their cardiac contractility is decreased with shorten rapid ejection phase.The myocardium is likely to remodel which leads to a slower heart rate and the decreased stroke volume may cause a low pulse pressure.Patients with CHD combined with HF are more likely to suffer from decreased function of aortic valve.The most common pulse conditions are infrequent pulse,uneven pulse,thready pulse,slippery pulse and taut pulse.Pulse graph parameters improved with moving average line method can provide more objective reference in studying features of pulse graph of CHD combined with HF.It is conducive to further promote the objectification of pulse diagnosis.