Comparison of remimazolam tosilate and propofol during induction and maintenance of general anesthesia in patients undergoing laparoscopic cholecystectomy: a prospective, single center, randomized controlled trial.

BACKGROUND: Remimazolam tosilate (RT) is a new, ultrashort-acting benzodiazepine. Here, we investigated the efficacy and safety of RT for general anesthesia in patients undergoing Laparoscopic Cholecystectomy (LC). METHODS: In this study, 122 patients undergoing laparoscopic cholecystectomy were randomly allocated to receive either remimazolam tosilate (Group RT) or propofol group (Group P). RT was administered as a slow bolus of 0.3 mg kg- 1 for induction, followed by 1.0-2.0 mg kg- 1 h- 1 for maintenance of general anesthesia. Propofol was started at 2 mg kg- 1 and followed by 4-10 mg kg- 1 h- 1 until the end of surgery. The primary outcome was the time to bispectral index (BIS) ≤ 60. The secondary outcome included the time to loss of consciousness (LoC), and the time to extubation. Adverse events were also assessed. RESULTS: A total of 112 patients were recruited for study participation. Among them, the time to BIS ≤ 60 in Group RT was longer than that in Group P (Group RT: 89.3 ± 10.7 s; Group P: 85.9 ± 9.7 s, P > 0.05). While the time to LoC comparing remimazolam and propofol showed no statistical significance (Group RT: 74.4 ± 10.3 s; Group P: 74.7 ± 9.3 s, P > 0.05). The time to extubation in Group RT was significantly longer than that in Group P (Group RT: 16.0 ± 2.6 min; Group P: 8.8 ± 4.3 min, P < 0.001). Remimazolam tosilate had more stable hemodynamics and a lower incidence of hypotension during general anesthesia. CONCLUSIONS: Remimazolam tosilate can be safely and effectively used for general anesthesia in patients undergoing Laparoscopic Cholecystectomy. It maintains stable hemodynamics during induction and maintenance of general anesthesia compared with propofol. Further studies are needed to validate the findings. TRIAL REGISTRATION: Chictr.org.cn ChiCTR2300071256 (date of registration: 09/05/2023).

Honokiol Prevents Intestinal Barrier Dysfunction in Mice with Severe Acute Pancreatitis and Inhibits JAK/STAT1 Pathway and Acetylation of HMGB1.

OBJECTIVE: To investigate the effect of honokiol (HON) and the role of high-mobility group protein B1 (HMGB1) on the pathogenesis of severe acute pancreatitis (SAP). METHODS: Thirty mice were numbered according to weight, and randomly divided into 5 groups using a random number table, including control, SAP, SAP and normal saline (SAP+NS), SAP and ethyl pyruvate (SAP+EP), or SAP+HON groups, 6 mice in each group. Samples of pancreas, intestine, and blood were collected 12 h after SAP model induction for examination of pathologic changes, immune function alterations by enzyme linked immunosorbent assay (ELISA), and Western blot. In vitro experiments, macrophages were divided into 5 groups, the control, lipopolysaccharide (LPS), LPS+DMSO (DMSO), LPS+anti-HMGB1 monoclonal antibody (mAb), and LPS+ HON groups. The tight connection level was determined by transmission electron microscopy and fluorescein isothiocyanate-labeled. The location and acetylation of HMGB1 were measured by Western blot. Finally, pyridone 6 and silencing signal transducer and activator of the transcription 1 (siSTAT1) combined with honokiol were added to determine whether the Janus kinase (JAK)/ STAT1 participated in the regulation of honokiol on HMGB1. The protein expression levels of HMGB1, JAK, and STAT1 were detected using Western blot. RESULTS: Mice with SAP had inflammatory injury in the pancreas, bleeding of intestinal tissues, and cells with disrupted histology. Mice in the SAP+HON group had significantly fewer pathological changes. Mice with SAP also had significant increases in the serum levels of amylase, lipase, HMGB1, tumor necrosis factor- α, interleukin-6, diamine oxidase, endotoxin-1, and procalcitonin. Mice in the SAP+HON group did not show these abnormalities (P<0.01). Studies of Caco-2 cells indicated that LPS increased the levels of occludin and claudin-1 as well as tight junction permeability, decreased the levels of junctional adhesion molecule C, and elevated intercellular permeability (P<0.01). HON treatment blocked these effects. Studies of macrophages indicated that LPS led to low nuclear levels of HMGB1, however, HON treatment increased the nuclear level of HMGB1 (P<0.01). HON treatment also inhibited the expressions of JAK1, JAK2, and STAT1 (P<0.01) and increased the acetylation of HMGB1 (P<0.05). CONCLUSION: HON prevented intestinal barrier dysfunction in SAP by inhibiting HMGB1 acetylation and JAK/STAT1 pathway.

Species Composition and Succession of Necrophagous Insects on Small Buried Baits in China.

The postburial interval (PBI) can be inferred by using necrophagous insects colonizing the buried corpse. In different seasons, the species composition and succession of necrophagous insects on swine carrion (0.5-0.75 kg) buried at the depths of 30 cm and 60 cm in a Populus alba var. pyramidalis (Bunge, 1854) (Salicales: Salicaceae) grove of Shenyang, China from 2017 to 2019 were investigated. A total of 21 species of necrophagous insects belonging to 5 orders, 17 families were collected. Among them, the species of Phoridae and Platystomatidae were dominant at burial depth of 30 cm and 60 cm in summer and autumn. The species composition and time of colonization of necrophagous insects on the buried baits varied with seasons. Platystoma mandschuricum (Enderlein, 1937) (Diptera: Platystomatidae) and Aleochara puberula (Klug, 1833) (Coleoptera: Staphylinidae), the first arriving insects in spring, occurred on the baits for the longest time, from early June to early December. This work could provide reference data for the PBI estimation in Shenyang and similar geographical areas.

Characterization and Phylogenetic Analysis of the Complete Mitochondrial Genomes of Two Tiny Necrophagous Phorid Flies, Metopina sagittata and Puliciphora borinquenensis (Diptera: Phoridae).

The mitochondrial genome is frequently used for species identification and phylogenetic studies. In this study, we first sequenced and annotated the complete mitochondrial genomes of two phorid species that are forensically important in buried or enclosed environments: Metopina sagittata (Liu) and Puliciphora borinquenensis (Wheeler). The complete mitochondrial genome sequences of M. sagittata and P. borinquenensis were 15,640 bp with an A+T content of 75.97% and 15,429 bp with an A+T content of 75.38%, respectively. Their circular genomes both contained 13 protein-coding genes (PCGs), 22 transfer RNA genes, 2 ribosomal RNA genes, and 1 control region located between rrnS and trnI which was 808 bp for M. sagittata and 746 bp for P. borinquenensis. All the PCGs of both species started with ATN codons except for cox1 which used TTG codon. In addition to the common stop codon TAA and TAG, the incomplete stop codon T was used in two PCGs (cox1 and nad4) of M. sagittata and five PCGs (cox1, cox2, cox3, nad5, and nad4) of P. borinquenensis. There were 3 and 10 mismatched base pairs in the tRNA secondary structures from M. sagittata and P. borinquenensis, respectively. Both maximum likelihood and Bayesian inference analyses indicated that Platypezidae and Phoridae are sister taxa. M. sagittata is closely related to P. borinquenensis within the subfamily Metopininae. This work enhances the databases of Phoridae genomes and contributes to the further study of species identification and phylogenetics of this family.

Intrapuparial Age Estimation of Forensically Important Dohrniphora cornuta (Diptera: Phoridae).

Dohrniphora cornuta (Bigot) is a forensically important phorid fly indoors and in burial environments. The determination of a minimum postmortem interval (PMImin) often relies on the determination of the age of the immatures. Although the larval development data of D. cornuta under different temperatures has been established, the intrapuparial stage which lasts for about half of the total immature development is scarce. In this study, we investigated the key morphological changes during intrapuparial development at constant temperatures (15, 18, 21, 24, 27, 30, 33, and 36°C), with an aim to estimate the intrapuparial age of D. cornuta. Puparia were sampled at 12-h (24, 27, 30, and 33°C), 24-h (18 and 21°C), and 48-h (15°C) intervals. The morphological developments within the puparium were analyzed using a stereomicroscope after the puparium was removed. The average minimum duration of intrapuparial stage was inversely related to temperature, ranging from 184.79 ± 3.00 h at 30°C to 1102.86 ± 25.55 h at 15°C for female, and 197.40 ± 4.12 h at 30°C to 1175.33 ± 18.55 h at 15°C for male. It did not develop at 36°C. Some morphological traits that changed during development within the puparium could be used as age markers. According to these changes, the intrapuparial stage of D. cornuta was divided into nine stages which could be used for both sexes. This study provides relatively systematic development data of D. cornuta intrapuparial for the estimation of PMImin in forensic entomology.

SnoN residue (1-366) attenuates hypertrophic scars through resistance to transforming growth factor-ß1-induced degradation.

Hypertrophic scars (HSs) are characterized by fibroblast hyperproliferation and excessive matrix deposition. During wound healing, transforming growth factor (TGF)-ß1/Smad signaling acts as a key regulator. As a transcriptional corepressor of TGF-ß1/Smads, SnoN is expressed at low levels in many fibrotic diseases due to TGF-ß1/Smad-induced degradation. SnoN residue (1-366; SR) is resistant to TGF-ß1-induced degradation. However, the expression and role of SR in HSs are unknown. Here, we inhibited TGF-ß1/Smad signaling via overexpression of SR to block fibroblast transdifferentiation, proliferation, and collagen deposition during HS formation. Our results showed that SnoN was downregulated in HS fibroblasts (HSFs) owing to TGF-ß1/Smad-induced degradation. Overexpression of SR in normal human dermal fibroblasts (NHDFs) and HSFs successfully blocked phosphorylation of Smad2 and Smad3, thereby inhibiting NHDF transdifferentiation and HSF proliferation and reducing type I collagen (ColI) and type III collagen (ColIII) production and secretion. In addition, we applied overexpressed full-length SnoN (SF) and SR to wound granulation tissue in a rabbit model of HSs. SR reduced wound scarring, improved collagen deposition and arrangement of scar tissue, and decreased mRNA and protein expression of ColI, ColIII, and α-smooth muscle actin (α-SMA) more effectively than SF in vivo. These results suggest that SR could be a promising therapy for the prevention of HS.

ASCL2 expression contributes to gastric tumor migration and invasion by downregulating miR223 and inducing EMT.

Achaete­scute homolog 2 (ASCL2), a basic helix­loop­helix transcription factor, serves an essential role in the maintenance of adult intestinal stem cells and the growth of gastric cancer (GC). However, the function of ASCL2 in the metastasis of GC is poorly understood. The present study aimed to evaluate the effect of ASCL2 expression on gastric tumor metastasis. ASCL2 protein expression was detected in 32 cases of gastric metastasis and its relevant primary tumors using western blotting and immunohistochemistry. The data suggested that the expression of ASCL2 was highest in metastatic tumors, among adjacent normal tissues, primary gastric tumors and gastric metastatic tumors. Furthermore, ASCL2­overexpressing GC cell lines MKN1­ASCL2 and SNU16­ASCL2 were established. An in vitro assay suggested that microRNA 223 (miR223) expression was downregulated following ASCL2 overexpression, and that the expression of the epithelium­associated protein E­cadherin was significantly decreased, while a series of mesenchyme­associated proteins, including zinc finger E­box­binding homeobox 1 (Zeb­1), twist­related protein 1, integrin, vimentin, 72 kDa type IV collagenase and matrix metalloproteinase­9 were upregulated in ASCL2­overexpressing cells. Overexpression of miR223 attenuated the epithelial­mesenchymal transition (EMT)­promoting effect induced by ASCL2 expression. In addition, the results of the chromatin immunoprecipitation and luciferase reporter gene assays indicated that ASCL2 was able to interact with the promoter of pre­miR223, and to inhibit the maturation of miR223, which may interact with the 3' untranslated region of Zeb­1 and inhibit EMT in tumor cells. The results of the present study demonstrated that ASCL2 was able to downregulate the expression level of miR223, contribute to EMT and promote gastric tumor metastasis, which indicated that ASCL2 may serve as a therapeutic target in the treatment of GC.

[Strategies of interventing complications in hemodialysis with classical prescriptions from clinical cases].

There are 400 thousand patients with long-term hemodialysis in China nowadays. Hemodialysis, as the most common alternative to renal replacement therapy, prolongs the life span of patients with end stage renal failure. However, there are still many complications of hemodialysis treatment. These complications reduce the quality of life of patients and may even endanger their life, and sometimes they are difficult to deal with. Classical prescriptions, commonly referred to as classical effective prescriptions in modern medicine, mainly indicating the formulas recorded in Treatise on Febrile Diseases and Synopsis of Golden Chamber, were relative to contemporary prescriptions emerging after Song and Yuan dynasties. Prescriptions corresponding to syndromes means one-to-one correspondence between syndromes and prescriptions, with many advantages and that is the key of clinical efficacy. Many complications of hemodialysis patients have typical clinical manifestations, which can match the syndromes corresponding to classical prescriptions, thus quickly relieving the symptoms of patients in clinical application. Six clinical cases of dialysis muscle spasm, disequilibrium syndrome, restless legs syndrome, uremic encephalopathy, post dialysis arrhythmia, and secondary hyperparathyroidism were used in this paper, to explore the intervention strategies for hemodialysis related complications.

Dachengqi Decoction Attenuates Intestinal Vascular Endothelial Injury in Severe Acute Pancreatitis in Vitro and in Vivo.

BACKGROUND/AIMS: Dachengqi decoction (DCQD) is a well-known traditional Chinese herbal drug with strong anti-inflammatory effects. Angiopoietin-1 (Ang-1) plays a vital role in maintaining the stability and integrity of the vascular wall and prevents vascular leakage due to inflammatory mediators. Our previous work found that DCQD protects against pancreatic injury in rats with severe acute pancreatitis (SAP). This study aims to investigate the effects of DCQD on intestinal endothelial damage in both damaged human umbilical vein endothelial cells (HUVECs) and SAP rats. METHODS: HUVECs were randomly divided into four groups: control group, TNF-α group, TNF-α plus Ang-1 group (Ang-1 group), and TNF-α plus DCQD group (DCQD group). Cells were incubated for 6 h, 12 h, and 24 h, before collection. The treatment concentration of DCQD was decided based on a Cell Counting Kit-8 (CCK-8) assay. The monolayer permeability of the HUVECs was assessed by measuring the transendothelial electrical resistance (TEER). Apoptosis was analyzed by flow cytometry. mRNA and protein expression of aquaporin 1 (AQP-1), matrix metalloproteinase 9 (MMP9), and junctional adhesion molecule-C (JAM-C) was evaluated by RT-PCR, immunocytofluorescence, and western blot. Forty male Sprague-Dawley rats were randomized into a control group, SAP group, SAP plus Ang-1 group (Ang-1 group), and SAP plus DCQD group (DCQD group). SAP was induced by intraperitoneal injection of cerulein and lipopolysaccharide (LPS), while the control group received 0.9% saline solution. Evans blue was injected through the penile vein and the rats were then sacrificed 12 h after modeling. Levels of serum amylase, TNF-α, IL-1ß, IL-2, and IL-6 were determined by using ELISA. Intestinal tissue was analysed by histology, and capillary permeability in the tissues was evaluated by Evans blue extravasation assay. Protein and mRNA expression of AQP-1, MMP9, and JAM-C were assessed by immunohistofluorescence, western blot, and RT-PCR. RESULTS: DCQD reduced the permeability of HUVEC induced by TNF-α in vitro. Furthermore, DCQD altered the mRNA and protein levels of JAM-C, MMP9, and AQP-1 in HUVECs after TNF-α induction. SAP intestinal injury induced by cerulein combined with lipopolysaccharides was concomitant with increased expression of JAM-C and MMP9, and reduced expression of AQP-1 in intestinal tissue. Pretreatment with DCQD attenuated SAP intestinal injury and lowered the levels of serum amylase, TNF-α, IL-1ß, IL-2, and IL-6 effectively. Our study demonstrated that DCQD decreased the expression of JAM-C and MMP9 and increased the expression of AQP-1 both in vitro and in vivo. CONCLUSION: DCQD can reduce capillary endothelial damage in acute pancreatitis-associated intestinal injury and the mechanism may be associated with the regulation of endothelial barrier function-associated proteins AQP-1, MMP9, and JAM-C.

Cantharidin Inhibits the Growth of Triple-Negative Breast Cancer Cells by Suppressing Autophagy and Inducing Apoptosis in Vitro and in Vivo.

BACKGROUND/AIMS: Cantharidin, a type of terpenoid secreted by the blister beetle Mylabris phalerata (Pallas), has attracted great attention in cancer therapy because of its potential anti-cancer activities. Here, we report the effects on apoptosis and autophagy in human triple-negative breast cancer (TNBC) cell lines after treatment with cantharidin and attempt to elucidate the underlying mechanisms. METHODS: MDA-MB-231 and MDA-MB-468 cells were treated with cantharidin and cell proliferation was examined using CCK-8 and clone formation assays. The expression of apoptosis- and autophagy-associated proteins was detected by western blotting. Cells were infected with lentivirus carrying the Beclin-1 gene, and MDA-MB-231-beclin1 (MB231-Bec) and MDA-MB-468-beclin-1(MB468-Bec) cells stably expressing Beclin-1 were established. Autophagic vacuoles in cells were observed with LC3 staining using fluorescence microscopy, and apoptotic cells were detected via flow cytometry. Tumor growth was assessed by subcutaneous inoculation of TNBC cells into BALB/c nude mice. RESULTS: Cantharidin inhibited the proliferation of MDA-MB-231 and MDA-MB-468 cells, and induced cell apoptosis. Cantharidin additionally inhibited the conversion of LC3 I to LC3 II and autophagosome formation by suppressing the expression of Beclin-1. Furthermore, overexpression of Beclin-1 in TNBC cells attenuated the cytotoxicity of cantharidin. In vivo, cantharidin inhibited the growth of MDA-MB-231 and MDA-MB-468 xenografts in nude mice by suppressing autophagy and inducing apoptosis, and Beclin-1 overexpression in TNBC cells reduced the efficacy of cantharidin. CONCLUSIONS: Cantharidin inhibits autophagy by suppressing Beclin-1 expression and inducing apoptosis of TNBC cells in vitro and in vivo, thereby representing a potential strategy for the treatment of TNBC.

Loss of the Opa interacting protein 5 inhibits breast cancer proliferation through miR-139-5p/NOTCH1 pathway.

Opa interacting protein 5 (OIP5) has been reported to be over-expressed in several kinds of human cancer. However, the biological function and clinical significance of OIP5 in human breast cancer remains unknown. In this study, we found that OIP5 was notably over-expressed in breast cancer tissues compared with their corresponding nontumorous tissues. Statistical analysis showed a significant correlation of OIP5 expression with advanced clinical stage. Ablation OIP5 inhibited the proliferation of breast cancer cells. OIP5 over-expression inhibited hsa-miR-139-5p expression, antagonized its functions and led to the de-repression of its endogenous target NOTCH1, which was a core oncogene in promoting breast cancer progression. Our results suggested that OIP5 is a potential diagnosis biomarker and therapeutic target for breast cancer.

SHCBP1 is over-expressed in breast cancer and is important in the proliferation and apoptosis of the human malignant breast cancer cell line.

BACKGROUND: SHC SH2-binding protein 1, a member of Src homolog and collagen homolog (Shc) family, has been recently identified in different contexts in unbiased screening assays. It has been reported to be over-expressed in several malignant cancers. METHODS: Immunohistochemistry of SHCBP1 on 128 breast cancer tissues and adjacent normal tissues were used to evaluate the prognostic significance of SHCBP1. Survival analyses were performed by Kaplan-Meier method. CRISPR/CAS9 method was used to knockout SHCBP1 expression. CRISPR/CAS9 technology was used to knockout SHCBP1 in 2 breast cancer cell lines. MTT assay, BrdU assay, colony formation assay, cell cycle assay and apoptosis analysis in MCF-7 and MDA-MB-231 cell lines were carried out to evaluate the effects of SHCBP1 on breast cancer in vitro. RESULTS: Immunohistochemical analysis revealed SHCBP1 was significantly up-regulated in breast cancer tissues compared with adjacent normal tissues (82 of 128, 64%). Over-expressed SHCBP1 was correlated with advanced clinical stage and poorer survival. Ablation of SHCBP1 inhibited the proliferation in vitro. SHCBP1 knockout increased cyclin-dependent kinase inhibitor p21, and decreased the Cyclin B1 and CDK1. CONCLUSION: Our study suggests SHCBP1 is dysregulated expressed in breast cancer and plays a critical role in cancer progression, which can be a potential prognosis predictor of breast cancer.

Early prediction of intestinal mucosal barrier function impairment by elevated serum procalcitonin in rats with severe acute pancreatitis.

OBJECTIVES: The aim of this study was to evaluate serum procalcitonin (PCT) levels as a prognostic indicator of intestinal barrier function impairment in rats with severe acute pancreatitis (SAP). METHODS: Thirty-six male Sprague Dawley rats were randomly grouped into SAP group (injected sodium taurocholate via biliopancreatic duct), Gln group (gavaged with glutamine after modeling), and control group. Blood, pancreatic, and terminal ileum tissues were obtained from the rats after 6 h of modeling. Serum amylase (Amy) levels were determined using an automatic biochemical detector, while endotoxin (ET), diamine oxidase (DAO), and PCT levels were measured by ELISA test. The pathology of pancreatic and small intestine tissues were observed. PCT protein expression in intestinal tissues were detected by immunohistochemistry and western blot. RESULT: Pancreatic and intestinal injuries in Gln group were significantly lower than SAP group. Serum amylase, DAO, and PCT levels in SAP and Gln groups differed greatly and were significantly higher than control group. Immuno-histochemistry and western blot results showed that PCT protein expression levels in small intestine tissues of SAP group were higher than Gln group and control group. Serum PCT levels had a significant correlation with serum endotoxin, DAO levels and intestinal mucosal injury scores. CONCLUSION: PCT expression in serum and intestinal tissues in SAP rats increased significantly in the early stages of SAP, and was closely related to the onset and degree of intestinal barrier function impairment. Thus, our results showed that measuring serum PCT can be used to predict intestinal mucosal barrier function impairment in SAP rats.

Cartilage Oligomeric Matrix Protein-Angiopoietin-1 Has a Protective Effect of Vascular Endothelial Barrier in Rat With Acute Necrotizing Pancreatitis.

OBJECTIVE: To investigate the protective effect of angiopoietin-1 (Ang-1) from capillary endothelial damage in rats with acute necrotizing pancreatitis (ANP). METHODS: 96 male Sprague-Dawley rats were randomly averaged and divided into control group, ANP group, Si-Ang-1 group, and COMP (cartilage oligomeric matrix protein)-Ang-1 group. Animals were killed at 6, 12, and 24 hours after molding. Levels of serum amylase, porcine endothelin 1, C-reactive protein, and Ang-1 were detected; histopathological changes in the pancreas were observed; capillary permeability and Ang-1 expression of the pancreatic tissue were detected by Evans Blue extravasation assay, immunohistochemistry, Western blot, and quantitative polymerase chain reaction. RESULTS: (1) Levels of serum amylase, C-reactive protein, and porcine endothelin-1 increased and level of Ang-1 decrease in the ANP group and Si-Ang-1 group compared with the control group, whereas COMP-Ang-1 group could improve the changes. (2) The order of pancreas pathological changes (mild to severe) is: control group, COMP-Ang-1 group, ANP group, and Si-Ang-1 group. (3) Capillary permeability of the pancreatic tissue in the COMP-Ang-1 group was lower than that in the ANP group. (4) Ang-1 mRNA and protein expression in the COMP-Ang-1 group was significantly higher than in the ANP group. CONCLUSIONS: COMP-Ang-1 can upregulate the expression of Ang-1 protein to promote angiogenesis and improve early inflammatory and pathological damage in ANP group.

Attenuation of peripheral regulatory T-cell suppression of skin-homing CD8⁺T cells in atopic dermatitis.

PURPOSE: Cutaneous lymphocyte-associated antigen (CLA)-expressing CD8⁺T cells have been known to play an important role in the pathogenesis of atopic dermatitis (AD). However, the mechanisms underlying the loss of self-tolerance remain unclear. Regulatory T cells (Tregs) play a key role in the development of homeostasis in the immune system. We, therefore, hypothesized that a reduced ability of Tregs to inhibit autologous CD8⁺CLA⁺T cells might be underlying mechanism in AD. MATERIALS AND METHODS: CD8⁺CLA⁺T cells and Tregs were obtained from the peripheral blood of AD patients and control volunteers. The frequencies of CD8⁺CLA⁺T cells were evaluated. The proliferative responses of CD8⁺CLA⁺T cells were assessed by flow cytometry, and the levels of transforming growth factor-ß1 (TGF-ß1) and interleukin-10 (IL-10) in culture supernatants were detected by enzyme-linked immunosorbent assay. RESULTS: Our results revealed higher frequency and increased expression of perforin and granzyme-B in peripheral CD8⁺CLA⁺T cells in AD, and lower inhibitory ability of Tregs on proliferation of CD8⁺CLA⁺T cells in AD. Meanwhile, the levels of TGF-ß1 produced by Tregs were significantly lower in AD, and anti-TGF-ß1 abolished such suppression. CONCLUSION: The attenuated inhibitory ability of Tregs on hyper-activated autologous CD8⁺CLA⁺T cells, mediated by TGF-ß1, plays an important role in the pathogenesis of AD.

Pupal age estimation of forensically important Megaselia scalaris (Loew) (Diptera: Phoridae).

In addition to causing myiasis in humans and animals, Megaselia scalaris (Loew) has been reported as a forensically important fly. The determination of a minimum postmortem interval (PMI) often relies on the determination of the age of the larvae and pupae. The pupal stage represents about 50% of the immature development time and the pupal age may therefore serve as an important tool in entomological minimum PMI estimation. The present study focuses on the key developmental processes during metamorphosis of M. scalaris pupae at different constant temperatures (18, 21, 24, 27, 30, 33 and 36°C). The average minimum duration of development from prepupa to adult emergence was inversely related to temperature, ranging from 170.65 ± 1.39 h at 33°C to 608.80 ± 13.26 h at 18°C. The pupa did not develop at 36°C. Some morphological features that changed during development within the puparium could be used as age markers. According to these changes, the pupal stage of M. scalaris was divided into 10 stages which could be used for both sexes.

Pupal age estimation of forensically important Megaselia spiracularis Schmitz (Diptera: Phoridae).

In addition to causing myiasis in humans, Megaselia spiracularis Schmitz has also been reported as a forensically important fly. The determination of a minimum post-mortem interval (PMI) often relies on the determination of the age of the larvae and pupae. The pupal stage represents about 50% of the immature development time and the pupal age may therefore serve as an important tool in entomological PMI estimation. The present study focusses on the key developmental processes during the metamorphosis of M. spiracularis pupae at different constant temperatures (21, 24, 27, 30, 33 and 36 °C). The average minimum duration of development from prepupa to adult emergence was inversely related to temperature, ranging from 177.10±1.65 h at 33 °C to 379.68±2.20 h at 21 °C. The pupa could not develop at 36 °C. Some morphological features that change within the puparium could be used as age markers. According to this, the pupal stage of M. spiracularis was divided into 11 stages which were fit for both sexes.

Morphology of immature stages of Dohrniphora cornuta (Bigot) (Diptera: Phoridae).

Morphology of all larval instars and puparium of Dohrniphora cornuta (Bigot), a most common phorid fly species indoors in China, is presented using scanning electron microscopy. The first instar larva was composed of 12 segments, each of segments 3-11 with six slender tubercles situated dorsally, dorsolaterally, and laterally in transverse row. These tubercles divided into two segments, of which the basal one was smooth, and the brush-shaped distal one was comprised of a cluster of fine spines. Antennae and maxillary palp complex were visible. Two slits could be seen at the posterior spiracle. Besides the presence of anterior spiracle, the tubercles of second instar became stouter than those of first instar and were covered by numerous long bristles from the base to top. The posterior spiracle contained four slits. Third larval instar was similar to second instar. The bubble membrane comprised of ≈120 globules with a pointed tip on their top presented at the segment 5 of third instar larvae. Puparia showed a retracted cephalic region and a pair of distinct pupal respiratory horns on the dorsum. The respiratory horns were long and bore numerous branches from base to apex. The apex of branch with two longitudinal slits was relatively broad and curled dorsally.

A study on morphology of immature stages of Diplonevra peregrina (Wiedemann) (Diptera: Phoridae).

Morphology of all larval instars and puparium of Diplonevra peregrina, a most common phorid fly species indoors in China, is presented using scanning electron microscopy. The first instar larva was composed of 12 segments, each of segments 3-11 with six spicate tubercles situated dorsally, dorsolaterally, and laterally in transverse row. The dorsal tubercles were much longer than the laterals and dorsolaterals. Antennae and maxillary palp complex were visible. The caudal segment was margined by six long, stout tubercles covered by numerous long bristles at the base through the apex. Two slits could be seen at the posterior spiracle. Besides the presence of anterior spiracle, the tubercles of second instar became more stout than those of first instar and were covered by numerous long bristles from the base to top. The posterior spiracle contained four slits. Third larval instar was similar to second instar. The bubble membrane comprised of clusters of small spines presented at the segment 5 of third instar larvae. Puparia showed a retracted cephalic region and a pair of pupal respiratory horns on the dorsum.

Down-regulation of aquaporin 1 in rats with experimental acute necrotizing pancreatitis.

OBJECTIVES: The objectives of this study were to investigate the expression of aquaporin 1 in capillary endothelial cells of rats with experimental acute necrotizing pancreatitis (ANP) and to explore its pathogenic role in capillary leak. METHODS: Sixty-four male Sprague-Dawley rats were randomly divided into control (n = 32) and ANP groups (n = 32). Eight rats in each group were killed at 3, 6, 12, and 18 hours after induction of experimental models. Quantity of ascites and levels of serum amylases were measured. Capillary permeability in pancreas, lung, and intestinal tissue was detected by Evans blue extravasation method. Aquaporin 1 expression in pancreas, lung, and intestinal tissue was detected by real-time polymerase chain reaction, immunohistochemical staining, and Western blot. RESULTS: Serum amylase level was significantly higher in ANP group than in controls (P < 0.05). Evans blue concentration in tissues in the ANP group was significantly higher than that in controls (P < 0.05). Aquaporin 1 mRNA and protein expressions in tissues were significantly less in the ANP group than in the controls (P < 0.05). CONCLUSIONS: The expression of aquaporin 1 was down-regulated in the pancreas, lung, and intestinal tissue of ANP rats, which could play an important role in the pathogenesis of capillary leak syndrome.