Abstract: Schistosomiasis, an important zoonotic parasitic disease, is one of the six major tropical diseases identified by WHO, and also one of the most important parasitic diseases for prevention and control in China. After more than 70 years of efforts, the prevention and control of schistosomiasis in China has made great achievements, and the current epidemic of schistosomiasis in China has entered an extremely low epidemic state, but the distribution base of the only intermediate host of schistosomiasis, Oncomelania hupensis, is still large. For now, the techniques used to monitor schistosomiasis have shortcomings such as time-consuming, laborious and low sensitivity, which cannot meet the current needs of China. Environmental DNA (eDNA) refers to DNA that can be extracted from environmental samples (such as soil, water or air) without isolating any target organisms, which is a complex mixture of genomic DNA and its degradation products from different organisms in the same environment. eDNA technology can reflect the community or species composition information in the ecosystem through DNA extraction and detection of environmental samples. Compared with traditional biological monitoring methods, eDNA technology has the advantages of high efficiency, high sensitivity and environmental friendliness. eDNA has been successfully used for the specific detection of Schistosoma mansoni, Schistosoma haematobium and Schistosoma japonicum. This paper reviews the current detection methods of eDNA, the application and technical limitations of eDNA technology in schistosomiasis monitoring, aiming to provide scientific reference for research in the field of schistosomiasis surveillance.

OBJECTIVETo determine the recognition of healthy medical students on deqi after acupuncture, reveal the qualitative and quantitative rules of deqi and understand whether these rules are the factors of the clinical application of acupuncture therapy.

METHODSThe class questionnaires were used for the investigation study on the understanding of deqi after acupuncture at Hegu (LI 4) or Zusanli (ST 36) in 86 healthy students in the clinical medicine class.

RESULTS(1) Deqi was a kind of complicated compound feelings, with many sensation qualities such as distending pain, distension and pain. (2) Deqi was a kind of mild and moderate sensations. In 10-score credit sys tem of Massachusetts General Hospital acupuncture sensation scale (MASS), the scores of distending pain (4.69 +/- 2.83), distension (4.39 +/- 2.91) and soreness and distension (3.93 +/- 2.93) were around 5 (moderate degree), the scores of stabbing pain (1.89 +/- 2.02) were around 2 (mild degree). (3) The differences in the quantitative scores were significant for stabbing pain, distending pain, distention, soreness and distention and the others before and after treatment (all P<0.05).

CONCLUSIONDeqi of acupuncture is the mild and moderate complicated sensations manifested as distension, soreness, pain and numbness. As the invasive therapy, the filiform needle puncture will bring a certain psychological impacts on the receptors. The subjective sensation is possibly the factor for the patients' selection of acupuncture treatment.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Adult ; China ; Female ; Humans ; Male ; Needles ; Qi ; Schools, Medical ; Sensation ; Young Adult

In order to optimize the ~(18)O labeling method, two key aspects, peptide dispersion and trypsin deac tivation were discussed o The addition of Rapigest SF in H_2~('8)O and microwave heating enhanced labeling efficiency of α-casein digested peptides(~(18)O/~(16)O) ratio >99%).Chemical modification with tris(2-carboxyeth yl) phosphine (TCEP) and iodoacetamide (IAA) resulted in trypsin deactivated completely.No significant back-exchange from ~(18)O to ~(16)O was observed after labeling in 6 days.The experiment result with peptide mixture from showed that the improved method could be effectively used to label protein and peptide.

ObjectiveTo explore the value of simvastatin in improving endothelial function in the patients with acute coronary syndromes in shorter time.Methods60 patients with acute coronary syndrome(acute myocardial infarction and unstable angina/non-ST elevation myocardial infarction) were randomized to be treated with placebo(n=30) or simvastatin 20 mg daily(n=30) for 3～5 d.At the admission and endpoint,Brachial ultrasound was used to measure endothelium-dependent flow-mediated dilatation(FMD) and response to endothelium-independent nitroglycerin. ResultsFMD was unchanged with placebo,but increased with simvasatin,from(2.65±2.95)% to(4.19±2.59)%(P=0.027).Responses to nitroglycerin were similar during the time course of the study in the 2 groups.The improvement of FMD was not correlated with the level of TC(R2=0.081,P=0.37),LDL-C(R2=0.056,P=0.46) or HDL-C(R2=0.073,P=0.40).ConclusionSimvastatin initiated early after acute coronary syndromes rapidly improves endothelial function in short course.No correlation has been detected between the pharmacological effects of simvastatin with the fall in TC and LDL-C.

BACKGROUND: One unit of umbilical cord blood does not have a sufficient number of peripheral blood stem cells to meet the requirements of transplantation in adults. One solution of this problem is their ex vivo expansion, which requires not only a longer time and higher culture conditions, but also easily leads to the differentiation of stern cells, thus affecting the effects of transplantation. OBJECTIVE: To transduce human interleukin-3(IL-3) gene into umbilical cord blood CD34" cells and to observe IL-3 expression. DESIGN, TIME AND SETTING: A cell-genomics in vitro experiment was performed in the Chengde Medical College in 2008. MATERIALS: Human peripheral blood mononuclear cells (PBMC) of healthy adult were provided by Chengde Blood Center, and umbilical cord blood was provided by Chengde Maternal and Child Care Hospital. Written informed consent was obtained from each donor. METHODS: Peripheral blood mononuclear cells were isolated by Ficoll gradient centrifugation and umbilical blood CD34+ cells were isolated using immunomagnetic beads method. IL-3 mRNA was extracted. IL-3 cDNA was synthesized by RT-PCR, and IL-3 cDNA was subcloned into eukaryotic expressing vector pcDNA3. In the experimental group, pcDNA3/IL-3 vectors were transduced into umbilical cord blood CD34+ cells, while in the control group, transfection was not performed. MAIN OUTCOME MEASURES: Detection of IL-3 level in umbilical cord blood CD34" cells suspension using ELISA kits. RESULTS: Theoretically, the amplified IL-3 cDNA was 616 bp, and actually, after agarose gel electrophoresis, the PCR products exhibited a strip with expected size under ultraviolet ray. Extraction of IL-3mRNA was successful and reversely transcripted cDNA was complete. A 616-bp inserted fragment was observed by agarose gel electrophoresis after double digestion with BamH Ⅰ and Xba Ⅰ, and it was the same as IL-3 sequence. Within 1-7 days after transfection, IL-3 level in the umbilical cord CD34+ cells suspension was significantly higher in the experimental group than in the control group (t = 3.46, P < 0.05). CONCLUSION: IL-3 cDNA was successfully cloned, and eukaryotic expressing plasmid pcDNA3/IL-3 that could be effectively expressed within short term in umbilical cord CD34+ cells was successfully constructed.

Objective : To evaluate the quality of neonatal inherited metabolic diseases screening in Chaoyang District, Beijing Municipality from 2012 to 2021, so as to provide insights into improvements in the screening quality and efficiency of neonatal inherited metabolic diseases. Methods: Data pertaining to screening of neonatal inherited metabolic disease in Chaoyang District from 2012 to 2021 were captured from Beijing Center for Neonatal Disease Screening. The percentage of screening, eligible rate of blood smears collection, re-examination rate of suspected cases, and definitive diagnosis of congenital hypothyroidism (CH), phenylketonuria (PKU) and congenital adrenal hyperplasia (CAH) were analyzed to evaluate the quality of neonatal inherited metabolic diseases screening in Chaoyang District. Results: There were 484 002 live neonates in Chaoyang District from 2012 to 2021, and 481 395 neonates were screened for inherited metabolic diseases, with a screening rate of 99.46% and 99.71% eligible rate of blood smears collection. A total of 4 305 suspected positive cases were screened, including 4 148 cases recalled for re-examinations, with a 96.35% re-examination rate of suspected cases, and the re-examination rates of CH, PKU and CAH were 96.37%, 96.79% and 95.65%, respectively. Totally 482 neonates were definitively diagnosed with inherited metabolic diseases, with an overall incidence rate of 1/999, and the incidence rates of CH (307 cases), hyperthyrotropinemia (103 cases), PKU (66 cases) and CAH (6 cases) were 1/1 568, 1/4 674, 1/7 294 and 1/20 233, respectively. Conclusions The screening rate and re-examination rate of neonatal inherited metabolic diseases was both more than 95% in Chaoyang District from 2012 to 2021. Improving the management of neonatal inherited metabolic diseases screening and the recall of suspected cases is required.

OBJECTIVETo observe enhanced effects of polypeptide extract from scorpion venom (PESV) combined Rapamycin on autophagy of H22 hepatoma cells in mice and to explore its possible mechanism.

METHODSThe H22 hepatocarcinoma cell suspension was subcutaneously inoculated into 40 Kunming mice. Then tumor-bearing mice were randomly divided into four groups, i.e., the control group,the high dose PESV group, the low dose PESV group, and the combination group (high dose PESV + Rapamycin), 10 in each group. Mice in high and dose PESV groups were administered with 20 mg/kg and 10 mg/kg PESV respectively by gastrogavage. Mice in the combination group were administered with 2 mg/kg rapamycin and 20 mg/kg PESV by gastrogavage. The intervention lasted for 14 successive days. The tumor volume was measured once every other day, the tumor growth curve was drawn, and then the tumor inhibitory rate calculated. Pathological changes of the tumor tissue were observed by HE staining. Protein expression levels of mammal target of rapamycin (mTOR), UNC-51-like kinase-1 (ULK1), microtubule-associated protein1 light chain3 (MAPILC3A), and Beclin1 were detected by immunohistochemical assay.

RESULTSThe growth of H22 hepatoma transplantation tumor was inhibited in high and low dose PESV groups and the combination group (P < 0.05). And there was statistical difference in tumor weight and tumor volume between the combination group and high and low dose PESV groups (P < 0.05). There was no statistical difference in tumor weight or tumor volume between the high dose PESV group and the low dose PESV group (P > 0.05). lmmunohistochemical assay showed that the protein expression of mTOR was higher, but protein expressions of ULK1, MAP1LC3A, Beclin1 were lower in the control group than in the rest 3 groups (P < 0.05, P < 0.01). Compared with the high dose PESV group, protein expressions of ULK1, MAP1LC3A, and Beclin1 were obviously lower (P < 0.05).

CONCLUSIONPESV combined Rapamycin might inhibit the development of H22 hepatoma transplantation tumor in mice possibly through inhibiting the activity of mTOR, enhancing expressions of ULK1, MAP1LC3A, and Beclin1.

Animals ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; therapeutic use ; Autophagy ; drug effects ; Carcinoma, Hepatocellular ; Cell Line, Tumor ; Liver Neoplasms ; Mice ; Neoplasm Transplantation ; Peptides ; Scorpion Venoms ; pharmacology ; therapeutic use ; Sirolimus ; pharmacology ; therapeutic use

Glutathione S-transferase (GST) gene, PcgstA was cloned from the penicillin producing strain Penicillium chrysogenum,which is important for understanding the industrial fermentation process. PcgstA gene has an open-reading-frame of 840 bp in length,which is interrupted by two introns. The deduced amino acid sequence shows about 50% identity to several characterized filamentous fungi GSTs. The recombinant PcGSTA in Escherichia coli were overexpressed and purified. Enzymatic assays showed that the recombinant PcGSTA had a specific activity with 1-chloro-2, 4-dinitrobenzene of (0.159±0.031) μmol/(min· mg). It was found that the expression level of PcgstA in the penicillin producing medium supplemented with phenylacetic acid, the side chain precursor of penicillin G, was significant down regulated than that in medium without phenylacetic acid. This result suggested that PcGST may be related to phenylacetic acid metabolism in the penicillin producing strain.

Objective To investigate the effect of fluvastatin (FLV) on the expression of β1 integrin in puromycin aminonucleoside (PAN)-treated podocytes and its mechanism.Methods Cultured human podocytes were divided into PAN,different concentrations of fluvastatin (1 × 10-8 to 1 × 10-5 mol/L),SOD,H2O21 groups respectively.Expressions of β1 integrin and reactive oxygen species (ROS) in podocytes were detected by Western blotting and DCFHDA (2' 7'-Dichlorofluoresecein 3' 6'-diacetate) respectively.The viability of podocyte was determined by MTT colorimetry.Results PAN and H2O2 significantly decreased the expression of β1 integrin and increased the synthesis of ROS in podocytes (P＜0.05respectively).Lower concentration fluvastatin or SOD treatment up-regulated β1 integrin and downregulated ROS of podocytes induced by PAN (P＜0.05 respectively).MTT revealed that lower podocyte viability was found in higher concentration fluvastatin,PAN and H2O2 groups.Lower concentration fluvastatin and SOD could protect podocytes against PAN.Conclusion Fluvastatin attenuates the injury of podocyte induced by PAN and increases the expression of β1 integrin,whose mechanism may be associated with the inhibition of the ROS activity.

Objective To explore the effect and mechanism of fluvastatin on the expression of fibronectin(FN) in human peritoneal mesothelial cells (HPMCs) induced by high-glucose peritoneal dialysate (HGPDS).Methods Cultured HPMCs were randomly divided into control,HGPDS,HGPDS plus GSK650394 10-5 mol/L (the competitive inhibitor of SGK1),different concentrations of fluvastatin,fluvastatin 10-6 mol/L and GSK650394 10-5 mol/L alone.The morphology change of HPMC was observed by light microscopy.The cellular viability was detected by MTT colorimetry.The mRNA and protein expressions of serum and glucocorticoid-inducible kinase 1 (SGK1) and FN were detected by RT-PCR,Western blotting or ELISA.Results After incubation with HGPDS,the cell morphology changed from typical cobblestone-like appearance to fibroblast-like appearance,and the cell viability was inhibited significantly (P＜0.05).Fluvastatin 10-6mol/L and GSK650394 could improved the cell morphology and the cell viability injured by HGPDS (P＜0.05).Compared with the normal control group,the mRNA and protein expressions of SGK1 and FN increased significantly in HPMC treated with HGPDS(P＜0.05).GSK650394 significantly decreased the high expression of SGK1 and FN (P＜0.05),also the fluvastatin had same effects as GSK650394 in dose-dependent manner (P＜0.05).Conclusions High-glucose peritoneal dialysate can increase FN expression in human peritoneal mesothelial cells,which can be attenuated by fluvastatin.The protective role of fluvastatin in HPMC may be partially achieved through the signal pathway of SGK1.