Selective degradation of PL2L60 by metabolic stresses­induced autophagy suppresses multi­cancer growth.

It has been reported that PL2L60 proteins, a product of PIWIL2 gene which might be activated by an intragenic promoter, could mediate a common pathway specifically for tumorigenesis. In the present study, it was further identified by using western blot assay that the PL2L60 proteins could be degraded in cancer cells through a mechanism of selective autophagy in response to oxidative stress. The PL2L60 was downregulated in various types of cancer cells under the hypoxic condition independently of HIF­1α, resulting in apoptosis of cancer cells. Inhibition of autophagy by small interfering RNA targeting of either Beclin­1 (BECN1) or Atg5 resulted in restoration of PL2L60 expression in hypoxic cancer cell. The hypoxic degradation of PL2L60 was also blocked by the attenuation of the autophagosome membrane protein Atg8/microtubule­associated protein 1 light chain 3 (LC3) or autophagy cargo protein p62 expression. Surprisingly, Immunofluorescence analysis demonstrated that LC3 could be directly bound to PL2L60 and was required for the transport of PL2L60 from the nucleus to the cytoplasm for lysosomal flux under basal or activated autophagy in cancer cells. Moreover, flow cytometric analysis displayed that knocking down of PL2L60 mRNA but not PIWIL2 mRNA effectively inhibited cancer cell proliferation and promoted apoptosis of cancer cells. The similar results were obtained from in vivo tumorigenic experiment, in which PL2L60 downregulation in necroptosis areas was confirmed by immunohistochemistry. These results suggested that various cancer could be suppressed by promoting autophagy. The present study revealed a key role of autophagic degradation of PL2L60 in hypoxia­induced cancer cell death, which could be used as a novel therapeutic target of cancer.

Band Gap and Defect Engineering Enhanced Scintillation from Ce3+-Doped Nanoglass Containing Mixed-Type Fluoride Nanocrystals.

Much can be learned from the research and development of scintillator crystals for improving the scintillation performance of glasses. Relying on the concept of "embedding crystalline order in glass", we have demonstrated that the scintillation properties of Ce3+-doped nanoglass composites (nano-GCs) can be optimized via the synergistic effects of Gd3+-sublattice sensitization and band-gap engineering. The nano-GCs host a large volume fraction of KYxGd1-xF4 mixed-type fluoride nanocrystals (NCs) and still retain reasonably good transparency at Ce3+-emitting wavelengths. The light yield of 3455 ± 20 ph/MeV is found, which is the largest value ever reported in fluoride NC-embedded nano-GCs. A comprehensive study is given on the highly selective doping of Ce3+ in the NCs and its positive effect on the scintillation properties. The favorable influence of the Y3+/Gd3+ mixing on the suppression of defects is accounted for by density functional theory and borne out experimentally. As a proof-of-concept, X-ray imaging with a good spatial resolution (7.9 lp/mm) is demonstrated by employing Ce3+-doped nano-GCs. The superior radiation hardness, repeatability, and thermal stability of the designed scintillators bode well for their long-term practical applications.

Preparation and Thermal Conductivity Enhancement of Boron Nitride Nano-Material PiG Composite.

With the improvement of the conversion efficiency of LED chip and fluorescent material and the increasing demand for high-brightness light sources, LED technology has begun to move toward the direction of high-power. However, there is a huge problem that high-power LED must face with a large amount of heat generated by high power causing a high temperature thermal decay or even thermal quenching of the fluorescent material in the device, resulting in a reduction of the luminous efficiency, color coordinates, color rendering index, light uniformity, and service life of LED. In order to solve this problem, fluorescent materials with high thermal stability and better heat dissipation were prepared to enhance their performance in high-power LED environments. A variety of boron nitride nanomaterials were prepared by the solid phase-gas phase method. By adjusting the ratio of boric acid to urea in the raw material, different BN nanoparticles and nanosheets were obtained. Moreover, the control of catalyst amount and synthesis temperature can be used to synthesize boron nitride nanotubes with various morphologies. By adding different morphologies and quantities of BN material in PiG (phosphor in glass), the mechanical strength, heat dissipation, and luminescent properties of the sheet can be effectively controlled. PiG prepared by adding the right number of nanotubes and nanosheets has higher quantum efficiency and better heat dissipation after being excited by high power LED.

Giant struma ovarii with pseudo-Meigs'syndrome and raised cancer antigen-125 levels: A case report.

BACKGROUND: Struma ovarii is a type of monodermal mature teratoma composed entirely or mainly of thyroid tissue, accounting for 1% to 3% of all ovarian teratomas and 0.3% to 1.0% of all ovarian tumors. Of which, struma ovarii with ascites and pleural effusion, called pseudo-Meigs'syndrome and raised cancer antigen-125 levels (CA 125) is even rarer. CASE SUMMARY: This paper reports the diagnosis and treatment of a patient of struma ovarii with pseudo-Meigs'syndrome, presenting with the clinical features of ovarian carcinoma: Complex pelvic mass, gross ascites, right pleural effusion and markedly elevated serum CA 125 levels. During the operation, a cystic-solid mass about 20 cm × 10 cm × 5 cm in the right adnexa and a solid mass with the size of 3 cm × 2 cm × 0.1 cm in the left ovary were observed. She underwent right adnexectomy and resection of the left ovarian mass and histopathology revealed a mature left-sided ovarian teratoma and struma ovarii of right adnexal mass. During 1-year follow-up, the patient recovered well, tumor markers and other indicators returned to normal. CONCLUSION: The diagnosis and treatment process of this case suggests that the clinical symptoms of struma ovarii with pseudo-Meigs'syndrome are lack specificity, which is easily misdiagnosed. Clinicians should improve the understanding of this disease, enhance the awareness of early screening, and improve the level of diagnosis and treatment.

Data-Driven Modelling and Control for Robot Needle Insertion in Deep Anterior Lamellar Keratoplasty.

Deep anterior lamellar keratoplasty (DALK) is a technique for cornea transplantation which is associated with reduced patient morbidity. DALK has been explored as a potential application of robot microsurgery because the small scales, fine control requirements, and difficulty of visualization make it very challenging for human surgeons to perform. We address the problem of modelling the small scale interactions between the surgical tool and the cornea tissue to improve the accuracy of needle insertion, since accurate placement within 5% of target depth has been associated with more reliable clinical outcomes. We develop a data-driven autoregressive dynamic model of the tool-tissue interaction and a model predictive controller to guide robot needle insertion. In an ex vivo model, our controller significantly improves the accuracy of needle positioning by more than 40% compared to prior methods.

Construction and theoretical insights into the ESIPT fluorescent probe for imaging formaldehyde in vitro and in vivo.

We report the first ESIPT-based probe ABTB, for the highly sensitive and selective imaging of formaldehyde (FA). The various theoretical calculations have been systematically performed, and clearly unravel the lighting mechanism of the fluorescent probe for FA. Additionally, the probe was successfully applied in monitoring endogenous FA in the brain of AD mice.

Optical Coherence Tomography Guided Robotic Needle Insertion for Deep Anterior Lamellar Keratoplasty.

OBJECTIVE: Deep anterior lamellar keratoplasty (DALK) significantly reduces the post-transplantation morbidity in patients eligible for partial-thickness cornea grafts. The popular "big bubble" technique for DALK is so challenging, however, that a significant fraction of corneal pneumodissection attempts fail for surgeons without extensive DALK-specific experience, even with previous-generation cross-sectional optical coherence tomography (OCT) guidance. We seek to develop robotic, volumetric OCT-guided technology capable of facilitating or automating the difficult needle insertion step in DALK. METHODS: Our system provides for real-time volumetric corneal imaging, segmentation, and tracking of the needle insertion to display feedback for surgeons and to generate needle insertion plans for robotic execution. We include a non-automatic mode for cooperative needle control for stabilization and tremor attenuation, and an automatic mode in which needle insertion plans are generated based on OCT tracking results and executed under surgeon hold-to-run control by the robot arm. We evaluated and compared freehand, volumetric OCT-guided, cooperative, and automatic needle insertion approaches in terms of perforation rate and final needle depth in an ex vivo human cornea model. RESULTS: Volumetric OCT visualization reduces cornea perforations and beneficially increases final needle depth in manual insertions by clinically significant amounts. Our automatic robotic needle insertion techniques meet or exceed surgeon performance in both needle placement and perforation rate. CONCLUSION: Volumetric OCT is a key enabler for surgeons, although robotic techniques can reliably replicate their performance. SIGNIFICANCE: Robotic needle control and volumetric OCT promise to improve outcomes in DALK.

A novel "AIE + ESIPT" near-infrared nanoprobe for the imaging of γ-glutamyl transpeptidase in living cells and the application in precision medicine.

γ-Glutamyl transpeptidase (GGT) has been reported as a biomarker of hepatocellular carcinoma (HCC), and its imaging is of great benefit for early detection in precise medicine as well as intraoperative navigation. Herein, we have designed and synthesized a novel near-infrared fluorescent probe coupled aggregation-induced emission (AIE) and excited-state intramolecular proton transfer (ESIPT) effect for the detection of GGT. Thanks to conjugated glutamate acid, this probe could be dispersed in aqueous solution and showed barely any fluorescence emission. Through a GGT-mediated enzymatic reaction, the aggregation state of the probe in aqueous solution was changed and an intramolecular hydrogen bond was formed, resulting in an enhanced fluorescence emission. An excellent linear relationship was observed and the concentration of GGT measured was in the range of 10-90 U L-1 with a limit of detection calculated at 2.9 U L-1. Its feasibility has been confirmed by detecting GGT in HepG2 cells with high specificity and long-term sustainability, satisfying clinical need. Moreover, this nanoprobe showed great potential for precise medicine guided surgery by realizing fluorescence imaging in human liver tumour tissue and distinguishing it from normal tissue. Thus, we supposed that our AIE coupled ESIPT fluorescent nanoprobe has great potential in the early detection of HCC, the selective fluorescence imaging of GGT positive cells during surgery and application in precision medicine.

ECRG4 acts as a tumor suppressor gene frequently hypermethylated in human breast cancer.

Human breast cancer is a malignant form of tumor with a relatively high mortality rate. Although esophageal cancer-related gene 4 (ECRG4) is thought to be a possible potent tumor suppressor gene that acts to suppress breast cancer, its precise role in this disease is not understood. Herein, we assess the correlation between ECRG4 expression and DNA methylation, probing the potential epigenetic regulation of ECRG4 in breast cancer. We analyzed ECRG4 promoter methylation via methylation-specific PCR (MSPCR), bisulfite sequencing, and a promoter reporter assay in human breast cancer cell lines and samples. Gene expression was assessed by quantitative real-time PCR (qPCR), while protein levels were assessed by Western blotting. CCK8 assays were used to quantify cell growth; Esophageal cancer-related gene 4 wound healing assays were used to assess cellular migration, while flow cytometry was used to assess apoptosis and cell cycle progression. Apoptosome formation was validated via CO-IP and Western blotting. We found that human breast cancer samples exhibited increased methylation of the ECRG4 promoter and decreased ECRG4 expression. Remarkably, the down-regulation of ECRG4 was highly associated with promoter methylation, and its expression could be re-activated via 5-aza-2'-deoxycytidine treatment to induce demethylation. ECRG4 overexpression impaired breast cancer cell proliferation and migration, and led to G0/G1 cell cycle phase arrest. Moreover, ECRG4 induced the formation of the Cytc/Apaf-1/caspase-9 apoptosome and promoted breast cancer cell apoptosis. ECRG4 is silenced in human breast cancer cells and cell lines, likely owing to promoter hypermethylation. ECRG4 may act as a tumor suppressor, inhibiting proliferation and migration, inducing G0/G1 phase arrest and apoptosis via the mitochondrial apoptotic pathway.

Diabetes Medication Use in Association with Survival among Patients of Breast, Colorectal, Lung, or Gastric Cancer / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Studies suggest that regular use of metformin may decrease cancer mortality. We investigated the association between diabetes medication use and cancer survival. MATERIALS AND METHODS: The current study includes 633 breast, 890 colorectal, 824 lung, and 543 gastric cancer cases identified from participants of two population-based cohort studies in Shanghai. Information on diabetes medication use was obtained by linking to electronic medical records. The associations between diabetes medication use (metformin, sulfonylureas, and insulin) and overall and cancer-specific survival were evaluated using time-dependent Cox proportional hazards models. RESULTS: After adjustment for clinical characteristics and treatment factors, use of metformin was associated with better overall survival among colorectal cancer patients (hazards ratio [HR], 0.55; 95% confidence interval [CI], 0.34 to 0.88) and for all four types of cancer combined (HR, 0.75; 95% CI, 0.57 to 0.98). Ever use of insulin was associated with worse survival for all cancer types combined (HR, 1.89; 95% CI, 1.57 to 2.29) and for the four cancer types individually. Similar associations were seen for diabetic patients. Sulfonylureas use was associated with worse overall survival for breast or gastric cancer (HR, 2.87; 95% CI, 1.22 to 6.80 and HR, 2.05; 95% CI, 1.09 to 3.84, respectively) among diabetic patients. Similar association patterns were observed between diabetes medication use and cancer-specific survival. CONCLUSION: Metformin was associated with improved survival among colorectal cancer cases, while insulin use was associated with worse survival among patients of four major cancers. Further investigation on the topic is needed given the potential translational impact of these findings.

Real-time corneal segmentation and 3D needle tracking in intrasurgical OCT.

Ophthalmic procedures demand precise surgical instrument control in depth, yet standard operating microscopes supply limited depth perception. Current commercial microscope-integrated optical coherence tomography partially meets this need with manually-positioned cross-sectional images that offer qualitative estimates of depth. In this work, we present methods for automatic quantitative depth measurement using real-time, two-surface corneal segmentation and needle tracking in OCT volumes. We then demonstrate these methods for guidance of ex vivo deep anterior lamellar keratoplasty (DALK) needle insertions. Surgeons using the output of these methods improved their ability to reach a target depth, and decreased their incidence of corneal perforations, both with statistical significance. We believe these methods could increase the success rate of DALK and thereby improve patient outcomes.

Enhanced Transparency of Rough Surface Sapphire by Surface Vitrifaction Process.

A novel, simple, and low-cost in situ surface vitrification method has been effectively developed to enhance the optical transparency of rough surface sapphire at UV-visible-IR regions. This method is to obtain a glass layer on the sapphire surface through vitrifaction process. The thickness, refractive index, components and transition temperature of the glass layer have been investigated and discussed respectively by XRD, DSC, SEM and EDS elemental analysis. The experimental results show that the vitrified sapphire has high transparency even after 1000 °C annealing at UV-visible-IR regions.

Effects of rehabilitation training on apoptosis of nerve cells and the recovery of neural and motor functions in rats with ischemic stroke through the PI3K/Akt and Nrf2/ARE signaling pathways.

This study was designed in order to investigate the effects between rehabilitation training on the apoptosis of nerve cells and the recovery of neural and motor functions of rats with ischemic stroke by way of the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) and nuclear factor E2-related factor 2/antioxidant responsive element (Nrf2/ARE) signaling pathways. In total, 110 healthy adult male Sprague-Dawley (SD) rats were selected in order to take part in this study. Ninety SD rats were used in order to establish the middle cerebral artery occlusion (MCAO), among which 80 rats were randomly assigned as part of the natural recovery, natural recovery+Rp-PI3K (the rats injected with PI3K/Akt inhibitor LY294002), rehabilitation training, and rehabilitation training+Rp-PI3K groups. Meanwhile, 20 rats were selected as part of the sham operation group. The neural and motor functions of these rats were evaluated using a balance beam test and the Bederson score. The mRNA expressions of PI3K, Akt, Nrf2 and HO-1 were measured using an RT-qPCR. The protein expressions of PI3K, p-PI3K, Akt, p-Akt, Nrf2 and HO-1 were also detected by using western blotting and the immunohistochemistry process. The cell cycle and cell apoptosis were detected by using a flow cytometry and TUNEL assay. The sham operation group exhibited lower neural and motor function scores than other groups. At the 7, 14, and 21 d marks of this study, the neural and motor function scores were increased in the natural recovery, natural recovery+Rp-PI3K, and rehabilitation training+Rp-PI3K groups in comparison with the rehabilitation training group but found to be decreased in the natural recovery group in comparison with the natural recovery+Rp-PI3K group. In comparison with the sham operation group, expressions of PI3K, Nrf2 and HO-1, and proportions of p-PI3K/PI3K and p-Akt/Akt were all higher in the natural recovery, rehabilitation training, and rehabilitation training+Rp-PI3K groups. Same trends were found in the rehabilitation training group in comparison with the natural recovery and rehabilitation training+Rp-PI3K groups, as well as in the natural recovery group in comparison with the natural recovery+Rp-PI3K group. In comparison with the sham operation and rehabilitation training groups, hippocampal nerve cells at G1 phase and the cells apoptosis were both elevated in the other three groups which were found to be decreased in the natural recovery group in comparison with the natural recovery+Rp-PI3K group. Our results indicated that the rehabilitation training can inhibit the apoptosis of nerve cells as well as promote the recovery of both neural Rehabilitation training in rats with IS.

Carnosic acid nanoparticles suppress liver ischemia/reperfusion injury by inhibition of ROS, Caspases and NF-κB signaling pathway in mice.

Living donor liver transplantation (LDLT) requires ischemia/reperfusion (I/R), which can lead to early graft injury. However, the detailed molecular mechanism of I/R injury remains unclear. Carnosic acid, as a phenolic diterpene with function of anti-inflammation, anti-cancer, anti-bacterial, anti-diabetic, as well as neuroprotective properties, is produced by many species from Lamiaceae family. Nanoparticulate drug delivery systems have been known to better the bioavailability of drugs on intranasal administration compared with only drug solutions. Administration of carnosic acid nanoparticles was thought to be sufficient to lead to considerable inhibition of liver injury progression induced by ischemia/reperfusion. In our study, liver ischemia/reperfusion injury was established successfully with C57BL/6 animal model. 10 and 20mg/kg carnosic acid nanoparticles were injected to mice for five days prior to ischemia. After liver ischemia/reperfusion, the levels of serum AST, ALT and APL were increased, which was attenuated by pre-treatment with carnosic acid nanoparticles. In addition, carnosic acid nanoparticles inhibited ROS production via its related signals regulation. And carnosic acid nanoparticles also suppressed the ischemia/reperfusion-induced up-regulation in the pro-apoptotic protein and mRNA levels of Bax, Cyto-c, Apaf-1 and Caspase-9/3 while increased ischemia/reperfusion-induced decrease of anti-apoptotic factor of Bcl-2. Further, ischemia/reperfusion-induced inflammation was also inhibited for carnosic acid nanoparticles administration via inactivating NF-κB signaling pathway, leading to down-regulation of pro-inflammatory cytokines releasing. In conclusion, our study suggested that carnosic acid nanoparticles protected against liver ischemia/reperfusion injury via its role of anti-oxidative, anti-apoptotic and anti-inflammatory bioactivity.

Plasma Metabolic Profile Determination in Young ST-segment Elevation Myocardial Infarction Patients with Ischemia and Reperfusion: Ultra-performance Liquid Chromatography and Mass Spectrometry for Pathway Analysis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>This study was to establish a disease differentiation model for ST-segment elevation myocardial infarction (STEMI) youth patients experiencing ischemia and reperfusion via ultra-performance liquid chromatography and mass spectrometry (UPLC/MS) platform, which searches for closely related characteristic metabolites and metabolic pathways to evaluate their predictive value in the prognosis after discharge.</p><p><b>METHODS</b>Forty-seven consecutive STEMI patients (23 patients under 45 years of age, referred to here as "youth," and 24 "elderly" patients) and 48 healthy control group members (24 youth, 24 elderly) were registered prospectively. The youth patients were required to provide a second blood draw during a follow-up visit one year after morbidity (n = 22, one lost). Characteristic metabolites and relative metabolic pathways were screened via UPLC/MS platform base on the Kyoto encyclopedia of genes and genomes (KEGG) and Human Metabolome Database. Receiver operating characteristic (ROC) curves were drawn to evaluate the predictive value of characteristic metabolites in the prognosis after discharge.</p><p><b>RESULTS</b>We successfully established an orthogonal partial least squares discriminated analysis model (R2X = 71.2%, R2Y = 79.6%, and Q2 = 55.9%) and screened out 24 ions; the sphingolipid metabolism pathway showed the most drastic change. The ROC curve analysis showed that ceramide [Cer(d18:0/16:0), Cer(t18:0/12:0)] and sphinganine in the sphingolipid pathway have high sensitivity and specificity on the prognosis related to major adverse cardiovascular events after youth patients were discharged. The area under curve (AUC) was 0.671, 0.750, and 0.711, respectively. A follow-up validation one year after morbidity showed corresponding AUC of 0.778, 0.833, and 0.806.</p><p><b>CONCLUSIONS</b>By analyzing the plasma metabolism of myocardial infarction patients, we successfully established a model that can distinguish two different factors simultaneously: pathological conditions and age. Sphingolipid metabolism is the top most altered pathway in young STEMI patients and as such may represent a valuable prognostic factor and potential therapeutic target.</p>

Direct Methylation of Amines with Carbon Dioxide and Molecular Hydrogen using Supported Gold Catalysts.

The N-methylation of amines with CO2 and H2 is an important step in the synthesis of bioactive compounds and chemical intermediates. The first heterogeneous Au catalyst is reported for this methylation reaction with good to excellent yields. The average turnover frequency (TOF) based on surface Au atoms is 45 h(-1) , which is the highest TOF value ever reported for the methylation of aniline with CO2 and H2 . Furthermore, the catalyst is tolerant toward a variety of amines, which includes aromatic, aliphatic, secondary, and primary amines. Preliminary mechanistic studies suggest that the N-alkyl formamide might be an intermediate in the N-methylation of amine process. Moreover, through a one-pot process, it is possible to convert primary amines, aldehydes, and CO2 into unsymmetrical tertiary amines with H2 as a reductant in the presence of the Au catalyst.

Left Lateral Sectionectomy of the Native Liver and Combined Living-Related Liver-Kidney Transplantation for Primary Hyperoxaluria Type 1.

Primary hyperoxaluria type I (PH1), the most severe form of primary hyperoxalurias, is a liver disease of the metabolic defect in glyoxylate detoxification that can be corrected by liver transplantation. A 21-year-old man presented to our center after 4 months of regular hemodialysis for kidney failure caused by nephrolithiasis. A diagnosis of PH1 was confirmed by mutations of the AGXT gene. Left lateral sectionectomy of the native liver was performed; and auxiliary partial orthotopic liver transplantation (APOLT) and kidney transplantation were carried out synchronously using a living donor. After transplantation, the patient's plasma oxalate and creatinine levels substantially decreased and the patient recovered well with good dual grafts function. APOLT and kidney transplantation can compensate the liver deficient in liver enzyme production and aid the renal elimination of oxalate, thus serving as an effective treatment option for patients with PH1. In conclusion, left lateral sectionectomy of the native liver and combined living-related liver-kidney transplantation can be a surgical option for PH1.

Re-evaluation of ABO gene polymorphisms detected in a genome-wide association study and risk of pancreatic ductal adenocarcinoma in a Chinese population / 癌症

Pancreatic cancer is a fatal malignancy with an increasing incidence in Shanghai, China. A genome-wide association study (GWAS) and other work have shown that ABO alleles are associated with pancreatic cancer risk. We conducted a population-based case-control study involving 256 patients with pathologically confirmed pancreatic ductal adenocarcinoma (PDAC) and 548 healthy controls in Shanghai, China, to assess the relationships between GWAS-identified ABO alleles and risk of PDAC. Carriers of the C allele of rs505922 had an increased cancer risk [adjusted odds ratio (OR) = 1.42, 95% confidence interval (CI): 1.02-1.98] compared to TT carriers. The T alleles of rs495828 and rs657152 were also significantly associated with an elevated cancer risk (adjusted OR = 1.58, 95% CI: 1.17-2.14; adjusted OR = 1.51, 95% CI: 1.09-2.10). The rs630014 variant was not associated with risk. We did not find any significant gene-environment interaction with cancer risk using a multifactor dimensionality reduction (MDR) method. Haplotype analysis also showed that the haplotype CTTC was associated with an increased risk of PDAC (adjusted OR = 1.46, 95% CI: 1.12-1.91) compared with haplotype TGGT. GWAS-identified ABO variants are thus also associated with risk of PDAC in the Chinese population.

[Geographic distribution and gene sequencing of Paragonimus westermani in some areas of Guangdong Province].

OBJECTIVE: To investigate the current distribution of Paragonimus westermani in Guangdong Province. METHOD: Snails and crabs collected from mountain streams in regional survey sites were dissected to detect cercarial and metacercarial infections of P. westermani. Domestic cats and dogs artificially infected with the collected metacercariae were also dissected to detect adult worms of P. westermani. The COI and ITS2 gene sequences of those adult worms were compared with those of known Paragonimus specimen deposited in the GenBank. RESULTS: All of the first intermediate hosts in five survey sites of Liangkou, Nankun, Mountain, Dadong, Muxi, Guowu, were identified as Semisulcospira libertina, whose cercariae infection rates were 0.33%, 0.15%, 0.058%, 0.10%, and 0.05%, respectively; the second intermediate hosts in above five sites were all identified as Sinopotamon denticulatum, whose metacercariae infection rates were 100%, 100%, 38.09%, 55.36%, and 65.26%, respectively. The numbers of metacercariae in the five sites were 79.4, 105.66, 9.16, 16.18, and 15.6 per positive crab, respectively, and 11.12, 7.87, 0.58, 0.69, and 0.85 per gram of crab, respectively. All the metacercariae were identical to those of P. westermani. Adult worms and eggs of P. westermani were found in both reservoir hosts of domestic cats and dogs infected artificially. By comparing the COI genes of five representative samples from each survey site with that of Paragonimus #AF219379.21, AF540958.1 from GenBank, we found out the homology to be 99%, 99%, 99%, 98%, and 99%, respectively. In addition, a comparison of the ITS2 gene sequences between the above five samples and Paragonimus #DQ836243.1, DQ351845.1, AB354217.1 from GenBank revealed 98%, 99%, 98%, 98%, and 98% gene homology, respectively. CONCLUSION: Two ultra-high and three high endemic areas of P. westermani are discovered in Guangdong Province. No obvious differences were found among the types of P. westermani in the above five endemic areas.