Pancreatic islet organoids-on-a-chip: how far have we gone?

Diabetes mellitus (DM) is a disease caused by dysfunction or disruption of pancreatic islets. The advent and development of microfluidic organoids-on-a-chip platforms have facilitated reproduce of complex and dynamic environment for tissue or organ development and complex disease processes. For the research and treatment of DM, the platforms have been widely used to investigate the physiology and pathophysiology of islets. In this review, we first highlight how pancreatic islet organoids-on-a-chip have improved the reproducibility of stem cell differentiation and organoid culture. We further discuss the efficiency of microfluidics in the functional evaluation of pancreatic islet organoids, such as single-islet-sensitivity detection, long-term real-time monitoring, and automatic glucose adjustment to provide relevant stimulation. Then, we present the applications of islet-on-a-chip technology in disease modeling, drug screening and cell replacement therapy. Finally, we summarize the development and challenges of islet-on-a-chip and discuss the prospects of future research.

The Density of Surface Coating Can Contribute to Different Antibacterial Activities of Gold Nanoparticles.

With the widespread use of antibiotics, the number of complex infection cases caused by unknown pathogens is increasing and novel antibiotics with tunable antibacterial spectra and low toxicity are highly desirable. Herein, we report that, by selecting thiol or amine, two groups with different binding affinities with gold, as anchoring groups, phenylboronic acid can be decorated on gold nanoparticles (AuNPs) with different densities, which contributes to Gram-selective antibacterial activities of the AuNPs. The AuNPs modified with amine- or thiol-tethered phenylboronic acids specifically bind to lipopolysaccharide (LPS, Gram-negative) or lipoteichoic acid (LTA, Gram-positive), respectively. By modifying AuNPs with different ratios of thiol- and amine-tethered phenylboronic acids, the resulting AuNPs show potent and tunable antibacterial activity. The AuNP-based antibacterial agents with optional Gram selectivity are promising for applications in personalized therapy.

Detection of Circulating Tumor Cells by Fluorescence Microspheres-Mediated Amplification.

Here we describe a fluorescent microspheres-based separation and analysis that enables the isolation of circulating tumor cells (CTCs) from whole blood of patients with metastatic cancer and the identification of isolated CTCs in situ without immunostaining. This approach uses antibody-functionalized fluorescent polystyrene (PS) microspheres that can selectively bind to CTCs. The binding of CTCs and fluorescent PS microspheres leads to the formation of complexes of CTCs and fluorescent PS microspheres, thereby the CTCs are size-amplified and labeled simultaneously. A pyramidal microcavity array (PMCA) is fabricated using microfabrication technology to create a precise microfilter structure with a high aspect ratio. The PMCA filter device can effectively isolate microspheres-labeled CTCs, while allow hematologic cells to deform and pass through. Using this approach, CTCs are isolated and identified in 15 of 18 patients with metastatic colorectal cancer. This approach will open new possibilities for CTCs isolation and identification and can serve a versatile platform to facilitate CTCs analysis in diverse biomedical applications.

The impact of meteorological parameters on the scrub typhus incidence in Baoshan City, western Yunnan, China.

Background: Scrub typhus has become widespread across various regions in China in recent decades, causing a considerable burden on residents. While meteorological variables significantly impact the spread of scrub typhus, there is insufficient quantitative evidence illustrating this association in known high-endemic areas. Methods: A distributed lag non-linear model was applied to explore the relationship between meteorological parameters and scrub typhus incidence from 2010 to 2019 in Baoshan City, western Yunnan Province, China. Results: High monthly mean (20°C) and maximum (30°C) temperatures were associated with a peak risk of scrub typhus in the current month. Higher minimum temperatures and higher relative humidity were followed by increasing cumulative risks over the ensuing 3 months. Higher precipitation was followed by increasing cumulative risk over the ensuing 2-month period, peaking at around 30 cm. Conclusion: The non-linear lag associations between meteorological parameters and scrub typhus incidence suggest that higher monthly minimum temperature and relative humidity could be associated with an increased risk of scrub typhus in the subsequent several months, while warm temperature is more likely to impact the occurrence of scrub typhus in the current month.

Genomic characterization of Rocahepevirus ratti hepatitis E virus genotype C1 in Yunnan province of China.

The Rocahepevirus ratti hepatitis E virus genotype C1 (HEV-C1) has been documented to infect humans. However, the understanding of HEV-C1 remains constrained. This study aims to determine the prevalence and genomic characteristics of HEV-C1 in small animals in Yunnan province of southwestern China. A total of 444 liver tissues were collected from animals covering the orders Rodentia, Soricomorpha, Scandentia and Erinaceomorpha in three regions in Yunnan. Then Paslahepevirus balayani and Rocahepevirus were examined using RT-qPCR. The detection rate of Rocahepevirus was 12.95 % (36/278) in animals of order Rodentia, with 14.77 % (35/237) in Rattus tanezumi and 33.33 % (1/3) in Niviventer fulvescens. No Paslahepevirus balayani was detected. Additionally, two full-length Rocahepevirus sequences (MSE-17 and LHK-54) and thirty-three partial ORF1 sequences were amplified and determined to be HEV-C1. MSE-17 and LHK-54 shared moderate nucleotide identity (78.9 %-80.3 %) with HEV-C1 isolated in rats and humans. The HEV-C1 isolated from Niviventer fulvescens demonstrated a 100 % nucleotide identity with that from Rattus tanezumi. The rat HEV-C1 sequences isolated in our study and other Asian HEV-C1 sequences were phylogenetically distant from those isolated in North America and Europe. Furthermore, the two full-length sequences isolated in our study had less amino acid substitutions in the motifs of RNA-dependent RNA polymerase domain (F204L and L238F), compared with other Asian sequences. In summary, HEV-C1 commonly spreads in rats in Yunnan province of China. Our findings suggest a spatially associated phylogeny, and potential cross-species transmission of HEV-C1.

FGF7 enhances the expression of ACE2 in human islet organoids aggravating SARS-CoV-2 infection.

The angiotensin-converting enzyme 2 (ACE2) is a primary cell surface viral binding receptor for SARS-CoV-2, so finding new regulatory molecules to modulate ACE2 expression levels is a promising strategy against COVID-19. In the current study, we utilized islet organoids derived from human embryonic stem cells (hESCs), animal models and COVID-19 patients to discover that fibroblast growth factor 7 (FGF7) enhances ACE2 expression within the islets, facilitating SARS-CoV-2 infection and resulting in impaired insulin secretion. Using hESC-derived islet organoids, we demonstrated that FGF7 interacts with FGF receptor 2 (FGFR2) and FGFR1 to upregulate ACE2 expression predominantly in ß cells. This upregulation increases both insulin secretion and susceptibility of ß cells to SARS-CoV-2 infection. Inhibiting FGFR counteracts the FGF7-induced ACE2 upregulation, subsequently reducing viral infection and replication in the islets. Furthermore, retrospective clinical data revealed that diabetic patients with severe COVID-19 symptoms exhibited elevated serum FGF7 levels compared to those with mild symptoms. Finally, animal experiments indicated that SARS-CoV-2 infection increased pancreatic FGF7 levels, resulting in a reduction of insulin concentrations in situ. Taken together, our research offers a potential regulatory strategy for ACE2 by controlling FGF7, thereby protecting islets from SARS-CoV-2 infection and preventing the progression of diabetes in the context of COVID-19.

Orientia tsutsugamushi Infection in Wild Small Mammals in Western Yunnan Province, China.

Small mammals can transmit and serve as a reservoir for Orientia tsutsugamushi (Ot) in nature by carrying infected mites. In Yunnan, one of China's main foci of scrub typhus, etiological evidence and genetic diversity for Ot is limited. A total of 2538 small mammals were captured seasonally from 2015 to 2016 in the three counties of Yunnan, and the spleen or liver tissue was examined for Ot based on 56 kDa nPCR. The overall prevalence of Ot was 1.77%, ranging from 0.26 to 9.09% across different species. The Gilliam strain was found in 35.6% (16/45) of the wild small mammals, followed by the Karp 11.1% (5/45) and TA763 (1/45) strains, the last of which was discovered in western Yunnan for the first time. In Lianghe, Ot infection rates in wild small mammals were higher than in the other two counties. The infection rates of Eothenomys miletus with Ot were highest in the three dominant species. Ot infection rates in wild small mammals were higher in Lianghe (1200-1400 m) and Yulong (2800-3000 m). These findings could provide research clues for further confirmation of scrub typhus foci in western Yunnan or other similar natural environments.

Molecular detection of Bartonella species in wild small mammals in western Yunnan Province, China.

Background: Small mammals serve as the main reservoir for Bartonella and as a proxy indicator of the potential risk of Bartonella transmission from nature to humans. They offer a valuable early warning for human infection. Nevertheless, geographical variations in the impact of the host on the occurrence of Bartonella infection are underestimated. This study was designed to investigate the infection characteristics of Bartonella and explore its species diversity in wild small mammals in western Yunnan Province, China. Methods: Wild small mammals were captured from Yulong, Jianchuan, and Lianghe counties in western Yunnan Province between 2015 and 2016. Real-time quantitative PCR (qPCR) was used to detect Bartonella infection, and the Bartonella species were identified by phylogenetic analysis. The factors associated with Bartonella infection in small mammals were analyzed by the Chi-square Test. Results: The prevalence of Bartonella in small mammals was 47.85% (768/1605). Lianghe County had the highest Bartonella infection rate, with 56.27% of the samples tested positive, followed by a rate of 50.91% was tested in Yulong County, and 39.97% in Jianchuan County (p < 0.001). Bartonella was detected positive in a total 25 small mammal species, with infection rates ranging from 2.17% to 100%. Niviventer fulvescens had the highest Bartonella infection rate. In comparison with the dominant small mammal species, Eothenomys mileyus had the lowest Bartonella infection rate than that in Apodemus chevrieri, Rattus tanezumi, and Apodemus draco (p < 0.001). Male small mammals had a higher infection rate than females (p < 0.05). The prevalence of Bartonella in small mammals during the summer season was higher compared to the other three seasons (p < 0.001). Woodland landscape had the highest Bartonella infection rate (p < 0.001). Bartonella rochalimae, B. japonica, B. tribocorum, B. washoensis, B. sylvatica, and B. rattimassiliensis were obtained from infected small mammals. Conclusion: This study showed a high prevalence of Bartonella was detected with various Bartonella species in small mammals in Yulong, Jianchuan, and Lianghe counties of western Yunnan Province. These findings hold significant scientific clues, providing valuable reference points for further research of Bartonella natural foci in Yunnan or other analogues environments.

[Detection and preliminary study of a family carrying a CCR5Δ32 deletional mutation].

OBJECTIVE: To investigate the frequencies of chemokine (C-C motif) receptor 5 gene (CCR5)Δ32 deletional mutation of in Han and Dai populations from Yunnan province. Immortalized cell lines were derived from a family carrying the CCR5Δ32 mutation. METHODS: Blood samples of 346 Han and 355 Dai individuals were collected for genotyping. The coding regions of CCR5 gene were amplified with PCR followed by agarose gel electrophoresis. Suspected mutations were verified with DNA sequencing. Immortalized cell lines were constructed by using Epstain Barr virus and cyclosporine A. The difference between the cell lines and original blood samples was verified with PCR. RESULTS: One ethnic Han individual was confirmed to be heterozygous for a deletional mutation by sequencing, which has led to discovery of a family with CCR5Δ32. Nine immortalized cell lines were established from this family, and no difference between the cell lines and original blood samples was detected by PCR. CONCLUSION: Together with previous reports, this study has indicated a significant difference in CCR5Δ32 among different ethnic groups in China. Established immortalized cell lines can also provide material for future research.

Genetic Evolution Analysis and Host Characteristics of Hantavirus in Yunnan Province, China.

For a long time, the epidemic situation of hemorrhagic fever with renal syndrome (HFRS) caused by hantavirus (HV) in Yunnan Province of China has been relatively severe. The molecular epidemiology and host characteristics of HV in Yunnan Province are still not completely clear, and the systematic and long-term investigation of the epidemic area is very limited. In this study, a total of 488 murine-shaped animals were captured in the three regions of Mile City, Mangshi City and Lianghe County in Yunnan Province, and then the type of HV was identified by multiplex real-time RT-PCR and sequenced. The results indicate that 2.46% of the murine-shaped animal specimens were infected with HV. A new subtype of Seoul virus (SEOV) was found in the rare rat species Rattus nitidus in Lianghe County, and the two strains of this new subtype were named YNLH-K40 and YNLH-K53. Through the phylogenetic analysis of this new subtype, it is shown that this new subtype is very similar to the type S5 of SEOV, which is previously described as the main cause for the high incidence of HFRS in Longquan City, Zhejiang Province, China. This new subtype is highly likely to cause human infection and disease. Therefore, in addition to further promoting the improvement of the HV gene database and strengthening the discovery and monitoring of the host animals in Yunnan Province, more attention should be paid to the pathogenic potential of the newly discovered HV type.

The Role of Calmodulin vs. Synaptotagmin in Exocytosis.

Exocytosis is a Ca2+-regulated process that requires the participation of Ca2+ sensors. In the 1980s, two classes of Ca2+-binding proteins were proposed as putative Ca2+ sensors: EF-hand protein calmodulin, and the C2 domain protein synaptotagmin. In the next few decades, numerous studies determined that in the final stage of membrane fusion triggered by a micromolar boost in the level of Ca2+, the low affinity Ca2+-binding protein synaptotagmin, especially synaptotagmin 1 and 2, acts as the primary Ca2+ sensor, whereas calmodulin is unlikely to be functional due to its high Ca2+ affinity. However, in the meantime emerging evidence has revealed that calmodulin is involved in the earlier exocytotic steps prior to fusion, such as vesicle trafficking, docking and priming by acting as a high affinity Ca2+ sensor activated at submicromolar level of Ca2+. Calmodulin directly interacts with multiple regulatory proteins involved in the regulation of exocytosis, including VAMP, myosin V, Munc13, synapsin, GAP43 and Rab3, and switches on key kinases, such as type II Ca2+/calmodulin-dependent protein kinase, to phosphorylate a series of exocytosis regulators, including syntaxin, synapsin, RIM and Ca2+ channels. Moreover, calmodulin interacts with synaptotagmin through either direct binding or indirect phosphorylation. In summary, calmodulin and synaptotagmin are Ca2+ sensors that play complementary roles throughout the process of exocytosis. In this review, we discuss the complementary roles that calmodulin and synaptotagmin play as Ca2+ sensors during exocytosis.

Enzyme-Regulated Peptide-Liquid Metal Hybrid Hydrogels as Cell Amber for Single-Cell Manipulation.

Current strategies to construct cell-based bioartificial tissues largely remain on a multicell level. Taking cell diversity into account, single-cell manipulation is urgently needed for delicate bioartificial tissue construction. Current single-cell isolation and profiling techniques involve invasive processes and thus are not applicable for single-cell manipulation. Here, we managed to fabricate peptide-liquid metal hybrid hydrogels as "cell ambers" which were suitable for single-cell isolation as well as further handling. The successful preparation of uniform liquid metal nanoparticles allowed the fabrication of peptide-liquid metal hydrogel with excellent recovery property upon mechanical destruction. The alkaline phosphatase-instructed supramolecular self-assembly process allowed the formation of microhydrogel post-filling in the PDMS template. The co-culture of the hydrogel precursor and mammalian cells realized the embedding of cells into elastic hydrogels which were the so-called cell ambers. The cell ambers turned out to be biocompatible and capable of supporting cell survival. Aided with the micro-operating system and a laser scanning confocal microscope, we could arrange these as-prepared 3D single-cell ambers into various patterns as desired. Our strategy provided the possibility to manipulate a single cell, which served as a prototype of cell architecture toward cell-based bioartificial tissue construction.

The relationship between fleas and small mammals in households of the Western Yunnan Province, China.

The Yunnan province has one of the most serious outbreaks of the plague epidemic in China. Small mammals and fleas are risk factors for the occurrence of plague in commensal plague foci. Understanding the relationship between fleas and small mammals will help control fleas and prevent the onset of the plague. Four hundred and twenty-one small mammals, belonging to 9 species, were captured. Of these, 170 small mammals (40.4%) were found infested with fleas. A total of 992 parasitic fleas (including 5 species) were collected. The number of Leptopsylla segnis and Xenopsylla cheopis accounted for 91.03% (903/992). The final multiple hurdle negative binomial regression model showed that when compared with Rattus tanezumi, the probability of flea infestation with Mus musculus as well as other host species decreased by 58% and 99%, respectively, while the number of flea infestations of the other host species increased by 4.71 folds. The probability of flea prevalence in adult hosts increased by 74%, while the number of fleas decreased by 76%. The number of flea infestations in small male mammals increased by 62%. The number of fleas in small mammals weighing more than 59 g has been multiplied by about 4. R. tanezumi is the predominant species in households in the west Yunnan province, while L.segnis and X. cheopis were dominant parasitic fleas. There is a strong relationship between the abundance of fleas and the characteristics of small mammals (e.g. Species, age, sex, and body weight).

Highly efficient capture of circulating tumor cells with low background signals by using pyramidal microcavity array.

This report demonstrates that a microfluidic device with integrated silicon filter exhibits outstanding capture efficiency and superior enrichment purity when employed to separate tumor cells from whole blood samples. We fabricate the silicon filter with pyramidal microcavity array (MCA) by microfabrication. We design the structure of the cavity to efficiently enrich tumor cells, while allowing hematologic cells to deform and pass through. The capture efficiency of MCF-7, SW620 and Hela cells spiked in 1 mL of whole blood are approximately 80%. Unwanted white blood cells (WBCs) trapped on the MCA are below 0.003%. In addition, this microfluidic device successfully identifies circulating tumor cells (CTCs) in 5 of 6 patients' blood samples, with a range of 5-86 CTCs per mL. These results reveal that the disposable microfluidic device can effectively enrich tumor cells with different sizes and various morphologies, while maintaining high capture efficiency and purity. Therefore, this label-free technique can serve as a versatile platform to facilitate CTCs analysis in diverse biochemical applications.

Predictors for presence and abundance of small mammals in households of villages endemic for commensal rodent plague in Yunnan Province, China.

BACKGROUND: Ninety-one rodent plague epidemics have occurred in Lianghe county, Yunnan Province, China, between 1990 and 2006. This study aimed to identify predictors for the presence and abundance of small mammals in households of villages endemic for rodent plague in Lianghe county. RESULTS: Rattus flavipectus and Suncus murinus were the two species captured in 110 households. Keeping cats decreased the number of captures of R. flavipectus by one to two thirds and the chance of reported small mammal sightings in houses by 60 to 80%. Food availability was associated with fewer captures. Keeping food in sacks decreased the small mammal captures, especially of S. murinus 4- to 8-fold. Vegetables grown around house and maize grown in the village reduced the captures of S. murinus and R. flavipectus by 73 and 45%, respectively. An outside toilet and garbage piles near the house each reduced R. flavipectus captures by 39 and 37%, respectively, while raising dogs and the presence of communal latrines in the village increased R. flavipectus captures by 76 and 110% but were without detectable effect on small mammal sightings. Location adjacent to other houses increased captures 2-fold but reduced the chance of sightings to about half. In addition, raising ducks increased the chance of sighting small mammals 2.7-fold. Even after adjusting for these variables, households of the Dai had higher captures than those of the Han and other ethnic groups. CONCLUSION: Both species captures were reduced by availability of species-specific foods in the environment, whereas other predictors for capture of the two species differed. Other than the beneficial effect of cats, there were also discrepancies between the effects on small mammal captures and those on sightings. These differences should be considered during the implementation and interpretation of small mammal surveys.

[Factors related to household rodent abundance in rodent-borne disease foci in western Yunnan].

OBJECTIVE: To analyze the factors related to the household abundance of rodents in rodent-borne disease foci in the western part of Yunnan province. METHODS: From July 2011 to October 2012, 800 households (20 households in 1 village) were randomly selected in 40 natural villages of 10 counties in western Yunnan where rodent borne disease was endemic to conduct a study on relationship between rodent abundance and environmental factors. Five cages were placed in each household for 3 consecutive nights to capture rodents. The rodent species were identified based on their morphological characteristics. The data on potential factors related to rodent abundance were collected through questionnaires and field observation. A dataset was established by using EpiData software and the analysis was performed with hurdle regression model under R software. RESULTS: A total of 421 rodents were captured in 800 households, belonging to 9 species, 6 genera, 2 families, 2 orders. Rattus tanezumi was the predominant species (66.03%). The final hurdle regression model showed that the probability of capturing rodents in the households where family member had high education level and the garbage was placed outside declined by 50%-68% ;The probability of capturing rodents in the households of Dai and Yi ethnic groups increased by 2.16-2.87 times;The probability of capturing rodents in the households where rodents were observed or vegetables grown near houses increased by 1.54-1.59 times;In the households where many rodents were believed to exist, the probability of capturing rodents and the number of rodents captured increased by 1.59 and 1.84 times respectively. The number of rodents captured in the houses with cement or tile floor increased by 3.62 times. CONCLUSION: The household abundance of rodents in the area in western Yunnan, where the rodent-borne disease survey was conducted, seemed to be closely related to the social economy status, human intervention and ecological environment. To control the abundance of rodents effectively, it is necessary to take these factors into consideration.

[Predictors for abundance of Rattus tanezumi in households of commensal rodent plague foci].

OBJECTIVE: To explore the predictors on the abundance of Rattus (R.) tanezumi in households of commensal rodent plague foci. METHODS: Thirty natural villages that experienced previous plague cases in Lianghe county, Yunnan province, were selected followed by random selection of 20 households in each village through computer technique. Live traps were set in households to capture small mammals which were then identified to species in the field according to their morphological features. Data on potential factors for abundance of R. tanezumi were collected through questionnaires and field observation and were coded and computerized using EpiData software and further analyzed by hurdle regression model under R software. RESULTS: A total of 166 rodents (133 R. tanezumi and 33 Suncus murinus) were captured. RESULTS: from final multilevel hurdle regression model showed that the likelihood of R. tanezumi captures increased by 1.67- to 2.76-fold in households belonged to Dai ethnic families that stored foodstuff in metal pail, often raising dogs, and having adjacent houses. The number of R. tanezumi captures increased by 2.18-fold in the villages where over 80% of the households would raise chickens. In the villages with communal latrine, the likelihood and the number of R. tanezumi capture increased 1.93-fold and 2.38-fold, respectively. While the likelihood of R. tanezumi captures would reduce by 45% - 61% in those households where there were cats and cattle being raised and maize grown in the village. The number of R. tanezumi captures would reduce by 63% in the households where there were outside toilet. CONCLUSION: The abundance of R. tanezumi seemed to be closely related to the ecological environment factors. Programs on plague control and prevention should relate to ecological factors that influencing the abundance of R. tanezumi.

Predictors for abundance of host flea and floor flea in households of villages with endemic commensal rodent plague, Yunnan Province, China.

BACKGROUND: From 1990 to 2006, fifty-five natural villages experienced at least one plague epidemic in Lianghe County, Yunnan Province, China. This study is aimed to document flea abundance and identify predictors in households of villages with endemic commensal rodent plague in Lianghe County. METHODS: Trappings were used to collect fleas and interviews were conducted to gather demography, environmental factors, and other relevant information. Multivariate hurdle negative binomial model was applied to identify predictors for flea abundance. RESULTS: A total of 344 fleas were collected on 101 small mammals (94 Rattus flavipectus and 7 Suncus murinus). R. flavipectus had higher flea prevalence and abundance than S. murinus, but the flea intensities did not differ significantly. A total of 315 floor fleas were captured in 104 households. Xenopsylla cheopis and Ctenocephalides felis felis were the predominant flea species on the host and the floor flea, respectively. The presence of small mammal faeces and R. flavipectus increased host flea prevalence odds 2.9- and 10-fold, respectively. Keeping a dog in the house increased floor flea prevalence odds 2-fold. Keeping cattle increased floor flea intensity by 153%. Villages with over 80% of houses raising chickens had increased prevalence odds and intensity of floor flea about 2.9- and 11.6-fold, respectively. The prevalence and intensity of floor flea in brick and wood houses were decreased by 60% and 90%, respectively. Flea prevalences of host and floor flea in the households that were adjacent to other houses were increased 7.4- and 2.2-fold, respectively. Houses with a paddy nearby decreased host flea intensity by 53%, while houses with an outside toilet increased host flea intensity by 125%. CONCLUSION: Rodent control alone may not be sufficient to control plague risk in these areas. In order to have successful results, plague control programs should pay attention to ecological and hygiene factors that influence flea populations.