Steinernema populi n. sp. (Panagrolaimomorpha, Steinernematidae), a new entomopathogenic nematode species from China.

Steinernema populi n. sp. was recovered by baiting from beneath poplar trees in China. Morphological and molecular features provided evidence for placing the new species into the Kushidai clade. The new species is characterized by the following morphological features: third-stage infective juveniles (IJ) with a body length of 1095 (973-1172) µm, a distance from the anterior end to excretory pore of 77 (70-86) µm and a tail length of 64 (55-72) µm. The Body length/Tail length (c) ratio and Anterior end to Excretory pore/ Tail length × 100 (E%) of S. populi n. sp. are substantially greater than those of all other 'Feltiae-Kushidai-Monticolum' group members. The first-generation males can be recognized by a spicule length of 66 (57-77) µm and a gubernaculum length of 46 (38-60) µm. The new species is further characterized by sequences of the internal transcribed spacer and partial 28S regions of the ribosomal DNA. Phylogenetic analyses show that Steinernema akhursti and Steinernema kushidai are the closest relatives to S. populi n. sp.

Virtual reality for acute and chronic pain management in adult patients: a narrative review.

Virtual reality is a computer-generated environment that immerses the user in an interactive artificial world. This ability to distract from reality has been utilised for the purposes of providing pain relief from noxious stimuli. As technology rapidly matures, there is potential for anaesthetists and pain physicians to incorporate virtual reality devices as non-pharmacological therapy in a multimodal pain management strategy. This systematic narrative review evaluates clinical studies that used virtual reality in adult patients for management of acute and chronic pain. A literature search found 690 citations, out of which 18 studies satisfied the inclusion criteria. Studies were assessed for quality using the Jadad and Nottingham-Ottawa Scales. Agreement on scores between independent assessors was 0.87 (95%CI 0.73-0.94). Studies investigated virtual reality use: intra-operatively; for labour analgesia; for wound dressing changes; and in multiple chronic pain conditions. Twelve studies showed reduced pain scores in acute or chronic pain with virtual reality therapy, five studies showed no superiority to control treatment arms and in one study, the virtual reality exposure group had a worsening of acute pain scores. Studies were heterogeneous in: methods; patient population; and type of virtual reality used. These limitations suggest the evidence-base in adult patients is currently immature and more rigorous studies are required to validate the use of virtual reality as a non-pharmacological adjunct in multimodal pain management.

Circular RNA circHIPK3 serves as a prognostic marker to promote chronic myeloid leukemia progression.

Increasing evidence demonstrate that circular RNAs (circRNAs) play critical role in regulation of gene expression, which participate in the pathogenesis of cancer, including chronic myeloid leukemia (CML). In this study, we aimed to investigate the expression profiling of circHIPK3 in CML. We found that circHIPK3 was significantly upregulated in peripheral blood mononuclear cells (PBMC) and serum samples from CML compared with healthy controls. High circHIPK3 expression predicted a poor outcome of CML patients. Further loss-function experiments suggested the oncogenic role of circHIPK3 in CML. Our findings provide insights on the role of circHIPK3 in the development and treatment of CML.

Enantioconvergent hydrolysis of m-nitrostyrene oxide at an elevated concentration by Phaseolus vulgaris epoxide hydrolase in the organic/aqueous two-phase system.

(R)-m-Nitrophenyl-1,2-ethanediol (m-NPED) is a versatile and highly value-added chiral building block for the synthesis of some bioactive compounds, such as (R)-Nifenalol. To efficiently produce (R)-m-NPED through the enantioconvergent hydrolysis of racemic (rac-) m-nitrostyrene oxide (m-NSO) using the whole resting cells of Escherichia coli/pCold-pveh2 intracellularly expressing PvEH2, an epoxide hydrolase from Phaseolus vulgaris, two reaction systems were investigated. In the Na2 HPO4 -NaH2 PO4 buffer (50 mmol l-1 , pH 7·0) system, merely 15 mmol l-1 rac-m-NSO was successfully subjected to enantioconvergent hydrolysis, producing (R)-m-NPED with 86·0% enantiomeric excess (eep ) and 177·6 mg l-1  h-1 space-time yield (STY). The experimental result indicated that there is inhibitory effect of rac-m-NSO at high concentration on PvEH2. To efficiently increase the concentration of rac-m-NSO and the STY of (R)-m-NPED, petroleum ether was first selected to construct an organic/aqueous two-phase system. Then, both the volume ratio (vo /vb ) of petroleum ether to phosphate buffer and the weight ratio (wc /ws ) of E. coli/pCold-pveh2 dry cells to rac-m-NSO were optimized as 2 : 8 and 5 : 1, respectively. In the optimized petroleum ether/phosphate buffer two-phase system, the enantioconvergent hydrolysis of rac-m-NSO at 40 mmol l-1 (6·6 mg ml-1 ) was carried out at 25°C for 12 h using 33·0 mg ml-1 vacuum freeze-dried cells of E. coli/pCold-pveh2, producing (R)-m-NPED with 87·4% eep , 82·3% yield and 502·4 mg l-1  h-1 STY. SIGNIFICANCE AND IMPACT OF THE STUDY: Epoxide hydrolases play a crucial role in producing enantiopure epoxides and/or vicinal diols. However, numerous biocatalytic reactions of organic compounds, such as epoxides, in aqueous phase suffered various restrictions. Herein, the enantioconvergent hydrolysis of rac-m-NSO in two reaction systems was investigated using the whole cells of Escherichia coli/pCold-pveh2. As a result, the concentration of rac-m-NSO and the space-time yield of (R)-m-NPED in organic/aqueous two-phase system were significantly increased, when compared with those in aqueous phase. To our knowledge, this is the first report about the production of (R)-m-NPED from rac-m-NSO at an elevated concentration by PvEH2 in the two-phase system.

[Analysis of follow-up results of chrono-chemotherapy or conventional chemotherapy combined with intensity modulated radiotherapy in locally advanced nasopharyngeal carcinoma].

Objective: To evaluate the long-term effect and safety of chrono-chemotherapy combined with intensity modulated radiotherapy (IMRT) in locally advanced nasopharyngeal carcinoma (NPC). Methods: 160 patients with locally advanced NPC were randomly divided into a chrono group and conventional group according to random number table. In the first stage, all patients underwent two cycles of induced chemotherapy, consisting of docetaxel, cisplatin and 5-Fu every 21 days. Notably, patients received chrono-moduated chemotherapy according to circadian rhythm in the chrono group, and conventional chemotherapy in the conventional group. Then, 21 days after the completion of first stage, three cycles of concurrent cisplatin chemotherapy every 21 days were given to all patients during IMRT. The median follow-up after the completion of radiotherapy was 31 months. Long-term side effects and the survival of patients were observed. Results: Patients in the chrono group had significantly lower rates of hearing loss (22.72%), dysphagia (0) and neck fibrosis (4.54%) compared with those in the conventional group (39.13%、8.69%, 15.94%, respectively, all P<0.05). Meanwhile, the 1- year overall survival rates (97.0% vs 92.8%), 3-year overall survival rates (80.3% vs 81.2%), 1-year progression free survival rates (95.5% vs 87.0%), 3-year progression free survival rates (71.2% vs 73.9%), 1-year locoregional relapse-free survival rates (97.0% vs 95.7%), 1-year locoregional relapse-free survival rates (92.4% vs 92.8%), 1-year distant metastasis-free survival rates (97.0% vs 98.6%) and 3-year distant metastasis-free survival rates (90.9% vs 91.3%) between the chrono group and the conventional group were not statistically significant (all P>0.05). Conclusions: Compared with conventional chemotherapy, chrono-chemotherapy combined with IMRT didn't affect long-term survival, but reducing the incidence of adverse events in patients with locally advanced NPC.

[Reevaluation of equivocal HER2 status detected by immunohistochemistry according to the 2019 guidelines for HER2 detection].

Objective To understand the effects and clinical significance of the 2019 guidelines for the human epidermal growth factor receptor 2 (HER2) detection. Methods: According to the 2014 guidelines, 548 cases of invasive breast cancer with equivocal HER2 (2+) detected by immunohistochemistry (IHC) in Taizhou Enze Medical Center, Zhejiang Province, China from 2013 to 2019 were selected. The results of IHC and HER2/CEPl7 double-probe were reevaluated and divided into groups according to the 2019 guidelines for the comparative analysis. Results: Among the 548 IHC HER2 (2+) invasive breast cancers, the number of positive, equivocal and negative cases for HER2 were 96 (17.52%), 81 (14.78%) and 371 (67.70%), respectively, according to the 2014 guidelines. However, according to the 2019 guidelines, 10 cases (1.82%) were reclassified as IHC 1+, 2 cases in the group 2 were reclassified as negative, and all the originally equivocal cases in group 4 were reclassified as negative. Finally, the total number of positive and negative cases for HER2 were 94 (17.15%) and 454 (82.85%), respectively. Conclusions: After applying the 2019 guidelines, the number of IHC 2+ cases decreases, and the positive rate for HER2 also decreases slightly due to the reevaluation change in groups 2 and 4, leading to reclassification of the cases that were deemed equivocal according to the 2014 guidelines. In general, the new 2019 guidelines are more reasonable and easier to use.

[The performance of visual photoscreening for Chinese preschool children aged 4 to 5 years].

Objective: To assess the accuracy of photoscreening for detecting refractive amblyopia risk factors (ARFs) in Chinese preschool children aged 4 to 5 years. Methods: A cross-sectional study. Comprehensive ocular examinations were conducted for preschool children in Nanjing, China from September to December, 2016. Photoscreening (Plusoptix A12C) was applied for refractive screening without cycloplegia. Voluntary children and children suspected of eyes abnormalities received cycloplegic retinoscopy (CR). Results of photoscreening and CR were compared using Wilcoxon signed rank test, and Bland-Altman plot were used to assess the agreement between the photoscreener and CR. According to the updated preschool vision screening guidelines from American Association for Pediatric Ophthalmology and Strabismus (AAPOS) in 2013, CR was adopted for identifying children with ARFs, which was considered as a golden standard. Based on the golden standard, the accuracy of 5 sets of referral criteria (including sensitivity standard, Matta/Silbert standard, AAPOS2013 standard, Alaska Blind Child Discovery standard, specificity standard) for photoscreener were tested. Receiver operating characteristics curves were constructed applied to evaluate the quality of the photoscreener in refractive ARFs detection and to find probably the best cut-off points. Results: In total, 1 986 children [mean age, (4.57±0.29) years] received comprehensive examinations, including 1 084 boys and 902 girls. The test ability of photoscreening was 99.04% (1 967/1 986) in the preschool children, and 96.56%(1 827/1 892) of the children got a reliable result within three screening attempts. In 538 children who had data of CR, refractive error of one child exceeded the upper limit of the photoscreener value setting, which was directly categorized as hyperopia, so in the end, 537 children were included to analyze the comparison between the two tests. The measurement values of photoscreening were lower than those of CR in sphere, cylinder and spherical equivalent [(0.75 (0.50, 1.25) D vs. 1.25 (1.00, 1.75) D, Z=-10.36, P<0.01; -0.50 (-0.75, -0.25) D vs. -0.25 (-0.75, 0.00) D, Z=-11.10, P<0.01; 0.63 (0.38, 0.88) D vs. 1.00 (0.75, 1.50) D, Z=-13.33, P<0.01]. The 95% limit of agreement cover rates between the photoscreening and CR in sphere, cylinder and spherical equivalent was 96.28% (517/537), 95.34% (512/537) and 96.65% (519/537), respectively. Based on the golden standard, 47 (8.74%) children had refractive ARFs, and the range of sensitivity, specificity, Youden index, positive predictive values and negative predictive values for detecting refractive ARFs of the 5 common used referral criteria was from 63.83% to 97.87%, from 53.36% to 97.56%, from 0.51 to 0.80, from 16.73% to 74.51% and from 96.57% to 99.62%, respectively. Considering particular refractive ARFs on the basis of the receiver operating characteristic curves, the optimal cut-off point for astigmatism was set at 1.38 D. Conclusion: Photoscreening could be an applicable tool to detect refractive ARFs in preschool children. (Chin J Ophthalmol, 2020, 56: 189-196).

Integrative transcriptomic and genomic analysis of odorant binding proteins and chemosensory proteins in aphids.

Odorant binding proteins (OBPs) and chemosensory proteins (CSPs) play essential roles in insect chemosensory recognition. Here, we identified nine OBPs and nine CSPs from the Myzus persicae transcriptome and genome. Genomic structure analysis showed that the number and length of the introns are much higher, and this appears to be a unique feature of aphid OBP genes. Three M. persicae OBP genes (OBP3/7/8) as well as CSP1/4/6, CSP2/9 and CSP5/8 are tandem arrayed in the genome. Phylogenetic analyses of five different aphid species suggest that aphid OBPs and CSPs are conserved in single copy across all aphids (with occasional losses), indicating that each OBP and CSP class evolved from a single gene in the common ancestor of aphids without subsequent duplication. Motif pattern analysis revealed that aphid OBP and CSP motifs are highly conserved, and this could suggest the conserved functions of aphid OBPs and CSPs. Three OBPs (MperOBP6/7/10) are expressed antennae specifically, and five OBPs (MperOBP2/4/5/8/9) are expressed antennae enriched, consistent with their putative olfactory roles. M. persicae CSPs showed much broader expression profiles in nonsensory organs than OBPs. None of the nine MperCSPs were found to be antennae specific, but five of them (MperCSP1/2/4/5/6) showed higher expression levels in the legs than in other tissues. MperCSP10 mainly expressed in the antennae and legs. The broad and diverse expression patterns of M. persicae CSPs suggest their multifunctions in olfactory perception, development and other processes.

Structural and biochemical evaluation of Ceratitis capitata odorant-binding protein 22 affinity for odorants involved in intersex communication.

In insects, odorant-binding proteins (OBPs) connect the peripheral sensory system to receptors of olfactory organs. Medfly Ceratitis capitata CcapObp22 shows 37% identity and close phylogenetic affinities with Drosophila melanogaster OBP69a/pheromone-binding protein related protein 1. The CcapObp22 gene is transcribed in the antennae and maxillary palps, suggesting an active role in olfaction. Here, we recombinantly produced CcapObp22, obtaining a 13.5 kDa protein capable of binding multiple strongly hydrophobic terpene compounds, including medfly male pheromone components. The highest binding affinity [half maximal effective concentration (EC50) = 0.48 µM] was to (E,E)-α-farnesene, one of the most abundant compounds in the male pheromone blend. This odorant was used in cocrystallization experiments, yielding the structure of CcapOBP22. The monomeric structure shows the typical OBP folding, constituted by six α-helical elements interconnected by three disulphide bridges. A C-terminal seventh α-helix constitutes the wall of a deep, L-shaped hydrophobic cavity. Analysis of the electron density in this cavity suggested trapping of farnesene in the crystal structure, although with partial occupancy. Superposition of the CcapOBP22 structure with related seven-helical OBPs highlights striking similarity in the organization of the C-terminal segment of these proteins. Collectively, our molecular and physiological data on medfly CcapOBP22 suggest its involvement in intersex olfactory communication.

[Effects of different fluid resuscitation methods on hemorheology in pigs during burn shock stage].

Objective: To investigate the impact of different methods of fluid resuscitation on hemorheology during burn shock stage. Methods: Twenty four miniature swines were randomly divided into four groups with 6 in each group (succinylated gelatin group, hydroxyethyl starch group, Parkland group and allogeneic plasma group). Severe burn shock model was established by burning miniature swine with napalm. Two hours after injury, succinylated gelatin, hydroxyethyl starch (130/0.4) and swine allogenic plasma were used as colloid (alternative colloid) in fluid resuscitation according to the burn shock fluid resuscitation formula which is commonly accepted in the field of Burns Surgery. In Parkland group, miniature swines received liquid recovery according to Parkland Formula. The vital signs before and within 48 h after burn were observed by Solar 8000i electrocardiomonitor during the process of transfusion. The infusion speed was adjusted based on the heart rate, blood pressure, urine volume and central venous pressure. The level of hematocrit (HCT), viscosity of plasma (ηp), index of rigidity (IR), red cell assembling index (RCA) and erythrocyte electrophoresis time (EFT) were measured at the time of pre-injury as well as 4, 8, 24 and 48 h post-injury and statistical analysis was performed. Results: HCT in hydroxyethyl starch group and Parkland group at 8 h post-injury were significantly higher than pre-injury [(0.395±0.047) vs (0.333±0.042), (0.379±0.026) vs (0.352±0.019)] (both P<0.05). And compared with pre-injury, HCT in hydroxyethyl starch (130/0.4) group at 48 h decreased significantly (0.232±0.021) vs (0.333±0.042) (P<0.05). HCT in Parkland group at 24, 48 h post-injury were lower than pre-injury [(0.277±0.021), (0.241±0.029) vs (0.352±0.019)] (both P<0.05). Compared with pre-injury, the levels of ηp in Parkland group decreased substantially at 4, 8 and 24 h post-injury [(1.61±0.07), (1.55±0.07) and (1.63±0.07) vs (1.73±0.04) mPa·s] (all P<0.05). Compared with allogeneic plasma group, IR decreased in succinylated gelatin group at 24, 48 h post-injury [(1.10±0.05 vs 1.26±0.07), (1.11±0.05 vs 1.32±0.05)](both P<0.05). RCA in succinylated gelatin group was significantly higher (both P<0.05) at 4 h (6.80±0.87) than pre-injury (5.92±0.43). RCA in hydroxyethyl starch group at 8 h post-injury (6.73±0.56) was significantly higher (both P<0.05) than pre-injury (6.03±0.53). Compared with pre-injury (17.3±1.3 s, 16.4±1.5 s), the levels of EFT in hydroxyethyl starch group (15.5±1.4 s) and Parkland group (13.4±1.2 s) decreased substantially at 48 h post-injury (both P<0.05). Compared with allogeneic plasma group, the level of EFT in succinylated gelatin group at 4 h post-injury (19.5±2.3 s) increased and decreased at 24 h post-injury (12.0±5.7 s) (both P<0.05). Conclusion: During swine burn shock stage, the hemorheological parameters of shock resuscitation with artificial colloid are more stable than those with Parkland formula resuscitation.

Identification and tissue expression profiling of candidate UDP-glycosyltransferase genes expressed in Holotrichia parallela motschulsky antennae.

It is difficult to control Holotrichia parallela Motschulsky with chemical insecticides due to the larvae's soil-living habit, thus the pest has caused great economic losses in agriculture. In addition, uridine diphosphate-glycosyltransferases (UGTs) catalyze the glycosylation process of a variety of small lipophilic molecules with sugars to produce water-soluble glycosides, and play multiple roles in detoxification, endobiotic modulation, and sequestration in an insect. Some UGTs were found specifically expressed in antennae of Drosophila melanogaster and Spodoptera littoralis, and glucurono-conjugated odorants could not elicit any olfactory signals, suggesting that the UGTs may play roles in odorant inactivation by biotransformation. In the current study, we performed a genome-wide analysis of the candidate UGT family in the dark black chafer, H. parallela. Based on a UGT gene signature and the similarity of these genes to UGT homologs from other organisms, 20 putative H. parallela UGT genes were identified. Bioinformatics analysis was used to predict sequence and structural features of H. parallela UGT proteins, and revealed important domains and residues involved in sugar donor binding and catalysis by comparison with human UGT2B7. Phylogenetic analysis of these 20 UGT protein sequences revealed eight major groups, including both order-specific and conserved groups, which are common to more than one order. Of these 20 UGT genes, HparUGT1265-1, HparUGT3119, and HparUGT8312 were highly (>100-fold change) expressed in antennae, suggesting a possible role in olfactory tissue, and most likely in odorant inactivation and olfactory processing. The remaining UGT genes were expressed in all tissues (head, thorax, abdomen, leg, and wing), indicating that these UGTs likely have different biological functions. This study provides the fundamental basis for determining the function of UGTs in a highly specialized olfactory organ, the H. parallela antenna.

[The analysis of clinical characteristics and disease activity in ankylosing spondylitis patients with hyperuircemia].

Objective: to investigate the clinical characteristics and disease activity in ankylosing spondylitis (AS) patients with hyperuricemia. Method: Laboratory tests, magnetic resonance imaging, disease activity and functional index of AS patients from Fudan University Zhongshan Hospital were collected. T test, square test were applied to investigate the difference between AS patients with hyperuricemia and AS patients with normal serum uric acid in clinical characteristics and disease activity in AS patients. Result: Among all the AS patients, 23.4% (22/94) patients accompanied with hyperuricemia. AS patients accompanied with hyperuricemia showed significant younger age (28.9±7.9 vs 35.8±11.1, P=0.002); higher serum alanine aminotransferase (33.6±23.6 vs 19.8±12.4, P=0.014), aspartate aminotransferase (23.8±8.7 vs 18.6±9.6, P=0.025) and creatinine (78.4±12.5 vs 69.8±13.3, P=0.009), lower Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) (11.2±8.8 vs 17.9±12.0, P=0.027) and Bath Ankylosing Spondylitis Functional Index (BASFI) (0.7±0.9 vs 1.8±2.2, P=0.002) compared with AS patients whose serum uric acid level is normal. Conclusion: AS Patients with younger age, impaired hepatic and renal funtion and lower scores in BASDAI and BASFI tend to accompany with hyperuricemia.

[Mild hypothermia combined with hydrogen sulfide alleviates hippocampal endoplasmic reticulum stress after global cerebral ischemia-reperfusion].

Objective: To investigate the effect of mild hypothermia combined with hydrogen sulfide on hippocampal endoplasmic reticulum stress (ERS) after global cerebral ischemia-reperfusion (I/R) injury. Methods: Sixty healthy male Sprague-Dawley rats, 8-10 week old, weighing 280-320 g, were randomly divided into 5 groups (n=12) using a random number table: sham operation group (group Sham), global cerebral I/R group (group I/R), hydrogen sulfide group (group H(2)S), mild hypothermia group (group MH) and hydrogen sulfide + mild hypothermia group (group H(2)S+ MH). Cardiac arrest was induced with transoesophageal cardiac pacing followed by cardiopulmonary resuscitation to establish the global cerebral I/R model. The administration regimen for sodium hydrosulfide (NaHS) was as follows: Sodium hydrosulfide was intraperitoneal injection as a bolus of 2.5 mg/kg immediately restoration of spontaneous circulation. The implementation of mild hypothermia: wipe the body surface of rats with ethanol immediately after restoration of spontaneous circulation, and reduce the rectal temperature to 32-34 ℃ within 15 min, and maintain 6 h with the ice bag. At 72 h of reperfusion, neurological deficit was scored, and the rats were sacrificed (Neurological Deficit Scores, NDS), the expression of glucose-regulated protein 78 (GRP78), CHOP and Caspase-12 were detected by Western blot. After reperfusion 72 h, the hippocampal tissue were removed and stained with haematoxylin and eosin to examine the pathological findings in hippocampal CA1 area (under microscope). The apoptosis rate of hippocampal CA1 area cells was detected by TUNEL staining and the apoptosis index was calculated. Results: The expression levels of endoplasmic reticulum stress marker, GRP78, CHOP and Caspase-12, were upregulates during the global cerebral ischemia reperfusion injury, indicating activation of severe endoplasmic reticulum stress. The GRP78 contents of Sham group, I/R group, H(2)S group, MH group and H(2)S+ MH group were as follows: GRP78: 0.11±0.03, 1.11±0.10, 0.67±0.09, 0.66±0.08, 0.48±0.04, CHOP contents: 0.16±0.03, 1.60±0.11, 1.39±0.09, 1.34±0.08, 1.13±0.09, Caspase-12 contents: 0.09±0.02, 0.87±0.08, 0.65±0.08, 0.59±0.06, 0.45±0.06, the differences were statistically significant (F=147.569, 264.983, 119.356, all P<0.01). The apoptosis index of Sham group, I/R group, H(2)S group, MH group and H(2)S+ MH group were as follows: (1.83±0.75)%, (53.17±4.62)%, (35.17±2.14)%, (32.67±2.25)%, (17.83±2.79)%, the differences were statistically significant (F=284.962, P<0.01). The neurological deficit scores of Sham group, I/R group, H(2)S group, MH group and H(2)S+ MH group were as follows: 0%, (76±9)%, (54±5)%, (47±6)%, (35±6)%, the differences were statistically significant(F=135.218, P<0.01). Conclusion: Mild hypothermia combined with hydrogen sulfide alleviates hippocampal endoplasmic reticulum stress after global cerebral ischemia-reperfusion, and the combined effect is better than that of a single application.

[Role of short-term starvation in alleviating hepatic ischemia-reperfusion injury in mice and possible mechanism of action].

Objective: To investigate the role of short-term starvation (STS) in alleviating hepatic ischemia-reperfusion injury in mice and possible mechanism of action. Methods: Wild-type male C57BL/6 mice aged 8 weeks were randomly divided into 75% hepatic ischemia-reperfusion injury group (IR group), STS+75% hepatic ischemia-reperfusion injury group (STS group), and sirtinol+STS+75% hepatic ischemia-reperfusion injury group (SIR group), using a random number table, and sham-operation groups (IR-Sham group, STS-Sham group, and SIR-Sham group) were also established. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured, and the histomorphological changes of the liver were observed, as well as the expression of Sirt1, LC3B, and P62 proteins in liver tissue and the results of LC3B fluorescence staining. An analysis of variance was used for comparison of data between multiple groups, and the t-test was used for comparison of data between two groups. Results: Compared with the IR group, the STS group had significant reductions in the serum levels of ALT (3 152.7 ± 735.6 U/L vs 8 414.2 ± 1 052.2 U/L, P < 0.01) and AST (3 577.0 ± 714.0 U/L vs 10 845.8 ± 1 145.7 U/L, P < 0.01) and significant alleviation of liver pathological injury (Suzuki score: 1.50±0.55 vs 3.50±0.55, P < 0.01). Compared with the STS group, the SIR group had significant increases in the serum levels of ALT (7 002.7 ± 1 485.2 U/L vs 3 152.7 ± 735.6 U/L, P < 0.01) and AST (8 980.7 ± 1 739.1 U/L vs 3 577.0 ± 714.0 U/L, P < 0.01) and significant exacerbation of liver pathological injury (Suzuki score: 3.33 ± 0.52 vs 1.50 ± 0.55, P < 0.01). Compared with the IR group and the IR-Sham group, the STS group and the STS-Sham group had significant increases in the mRNA and protein expression of Sirt1 and the protein expression of LC3B and a significant reduction in the protein expression of P62, as well as a significant increase in the percentage of LC3B-positive cells in liver tissue (22.83% ± 5.19% / 22.17% ± 4.83% vs 10.16% ± 3.06% / 10.83% ± 1.94%, both P < 0.01). Compared with the STS group and the STS-Sham group, the SIR group and the SIR-Sham group had significant reductions in the expression of Sirt1 and LC3B proteins and a significant increase in the expression of P62 protein, as well as a significant reduction in the percentage of LC3B-positive cells in liver tissue (11.83% ± 9.24% / 14.67% ± 4.68% vs 22.83% ± 5.19% / 22.17% ± 4.83%, both P < 0.01). Conclusion: STS can effectively alleviate hepatic ischemia-reperfusion injury, and its protective effect may be associated with increasing the expression of Sirt1, inducing and promoting hepatocyte autophagy, and reducing hepatocyte death.

Exacerbation of spontaneous autoimmune nephritis following regulatory T cell depletion in B cell lymphoma 2-interacting mediator knock-out mice.

Regulatory T cells (Tregs ) have been recognized as central mediators for maintaining peripheral tolerance and limiting autoimmune diseases. The loss of Tregs or their function has been associated with exacerbation of autoimmune disease. However, the temporary loss of Tregs in the chronic spontaneous disease model has not been investigated. In this study, we evaluated the role of Tregs in a novel chronic spontaneous glomerulonephritis model of B cell lymphoma 2-interacting mediator (Bim) knock-out mice by transient depleting Tregs . Bim is a pro-apoptotic member of the B cell lymphoma 2 (Bcl-2) family. Bim knock-out (Bim-/- ) mice fail to delete autoreactive T cells in thymus, leading to chronic spontaneous autoimmune kidney disease. We found that Treg depletion in Bim-/- mice exacerbated the kidney injury with increased proteinuria, impaired kidney function, weight loss and greater histological injury compared with wild-type mice. There was a significant increase in interstitial infiltrate of inflammatory cells, antibody deposition and tubular damage. Furthermore, the serum levels of cytokines interleukin (IL)-2, IL-4, IL-6, IL-10, IL-17α, interferon (IFN)-γ and tumour necrosis factor (TNF)-α were increased significantly after Treg depletion in Bim-/- mice. This study demonstrates that transient depletion of Tregs leads to enhanced self-reactive T effector cell function followed by exacerbation of kidney disease in the chronic spontaneous kidney disease model of Bim-deficient mice.

[Effects of resuscitation by different formulation on oxygen metabolism during shock stage of burn injury in swine].

OBJECTIVE: To explore the effects of different fluid resuscitation regimens on oxygen metabolism during shock stage of burn injury in swine. METHODS: Twelve Bama miniature swines were divided into crystal and colloid group (Group 1) and Parkland group (Group 2) according to the random number table. The swine models of burns shock were established. The fluid resuscitation was begun at post injury hour (PIH) 2 according to Chinese formulation or Parkland's formulation, respectively. The blood pressure, urine volume, heart rate, central venous pressure (CVP), pulmonary capillary wedge pressure (PCWP) were recorded. The liquid volume was calculated at the first and second PIH 24. The changes in oxygen delivery (DO2), oxygen consumption (VO2), oxygen extraction (O2Ext) and D-lactate (D-LA) were determined before injury and at PIH 4, 8, 24, and 48. Statistical analyses were performed. RESULTS: There were no statistical differences between the two groups in blood pressure, urine volume, heart rate, CVP, PCWP in every interval (all P>0.05). The resuscitation liquid volume in the two groups during the first and second PIH 24 conformed to the domestic consensus. The VO2 at PIH 8 was significantly higher than that of pre-burn in both groups [(190±29) vs (83±42) L·min(-1)·m(-2,) (149±33) vs (85±15) L·min(-1)·m(-2,) both P<0.05], and the VO2 at PIH 8 was significantly higher in Group 1 than that in Group 2 (P<0.05). The DO2 at PIH 24 in Group 1 was significantly lower than that in Group 2 [(686±72) vs (853±81) L·min(-1)·m(-2,) P<0.05]. There were no significant differences between the two groups in O2Ext at any time points (all P>0.05). The D-LA at PIH 8 was significantly higher in Group 1 than that in Group 2 [(53±4) vs (45±6) mmol/L, P<0.05]. CONCLUSION: There are no significant differences in the resuscitation effects of the crystal and colloid resuscitation regimen and Parkland's formulation on oxygen metabolism during shock stage of burn injury in swine.

Metabolic response of LLC xenografted mice to oxythiamine, as measured by [¹H] NMR spectroscopy.

Oxythiamine (OT) has been proven to be a potential anticancer drug. With the help of NMR-based metabonomics, we studied the metabolic changes within tumor-bearing mice with different levels of OT administration using a C57BL/6 mouse Lewis lung carcinoma tumor transplantation model. We administered different concentrations of OT (75, 150, 300, and 600 mg∙kg(-1)∙day(-1)) to the mice orally for 2 weeks, recorded animal weights and tumor volumes, sacrificed the animals, and collected blood and tumor mass samples for nuclear magnetic resonance determination. Compared with the findings for the control (untreated) group, the tumor weights and volumes of the 150, 300, and 600 mg∙kg-1∙day-1 groups decreased with no difference among these OT groups. A large metabolite difference was observed in plasma metabolites between the blank and control groups, which indicated the success of the tumor-bearing model. The metabolites in tumor associated with thiamine-dependent enzymes (TDEs) underwent considerable change between the OT and control groups, exhibiting concentration dependence and enzyme specificity. The restriction of TDEs by OT may be a major mechanism underlying its anticancer effect. The role of OT as a potential anticancer drug and a dehydrogenase inhibitor should therefore be taken into consideration in future tumor research.

The antenna-specific odorant-binding protein AlinOBP13 of the alfalfa plant bug Adelphocoris lineolatus is expressed specifically in basiconic sensilla and has high binding affinity to terpenoids.

Odorant-binding proteins (OBPs) are crucial in the olfactory pathway of insects. In the present study, the antenna-enriched OBP AlinOBP13 was investigated because of its potential contribution to the peripheral olfactory perception in the alfalfa plant bug Adelphocoris lineolatus. The results of quantitative reverse transcriptase-PCR showed that the transcript level of AlinOBP13 was higher in the adult stage than in the nymph stages. The transcript levels of AlinOBP13 in the male and female antennae significantly increased after 4 and 8 h of starvation, respectively. Fine ultrastructures of different types of chemosensilla in both female and male antennae were investigated using transmission electron microscopy and immunocytochemical labelling. The results revealed that the anti-AlinOBP13 antiserum strongly and specifically labelled short basiconic sensilla; this antiserum was restricted to the inner lumen and the cavities below the sensillum base of the sensilla. By contrast, multiporous sensilla trichodea, medium long sensilla basiconica, and aporous sensilla chaetica were not labelled. The present study is the first to report an OBP showing specific expression in the short basiconic sensilla of a member of the Hemipteran species. The results of a fluorescence displacement binding assay indicated that recombinant AlinOBP13 showed a more specific binding preference to terpenoids than to sex pheromones and other classes of chemicals. This binding ability was dramatically affected by pH; higher binding affinities were displayed at pH 10.0 than at pH 7.4 and 5.0. In addition, the results of dose-dependent electroantennogram recordings from the antennae showed that both female and male adult bugs responded to the terpenoids tested, suggesting an apparent physiological relevance of AlinOBP13 in A. lineolatus chemoreception. The results of this study suggest that AlinOBP13 functions as a specific carrier of terpenoids and provide insights into the mechanism of A. lineolatus in response to green volatiles.

Utilization of excess NADH in 2,3-butanediol-deficient Klebsiella pneumoniae for 1,3-propanediol production.

AIMS: To utilize excess NADH for 1,3-propanediol production by 2,3-butanediol-deficient mutants, the effect of dhaT overexpression in two distinct 2,3-butanediol-deficient mutants was investigated. METHODS AND RESULTS: Two 2,3-butanediol-deficient mutants, KG1-3 (blocking of the 2,3-butanediol pathway only) and KG1-5 (blocking of both of 2,3-butanediol and lactate pathways) were constructed. Our results showed that although the intracellular redox balance (NADH/NAD(+)) was extremely high at the end of fermentation for both mutants, the status of intracellular redox in KG1-5 was maintained at a normal level following the first stage of fermentation. Analysis of cell growth and metabolite formation confirmed the inhibition of excess lactate in 2,3-butanediol pathway-deficient mutants. Furthermore, dhaT was overexpressed in two 2,3-butanediol-deficient mutants (KG1-3T and KG1-5T). In KG1-5T, the intracellular redox balance was restored to normal and 1,3-propanediol production increased. The yield of 1,3-propanediol from glycerol in KG1-5T was also restored to a normal level of 0·6. CONCLUSIONS: The excess NADH in both the 2,3-butanediol- and lactate-deficient mutants can be used by overexpresstion of dhaT. SIGNIFICANCE AND IMPACT OF STUDY: The metabolic flux tended to increase lactate production by the abolishment of the 2,3-butanediol pathway in Klebsiella pneumoniae, and the high accumulation of lactate prevented the cell from using excess NADH, thereby inhibiting cell growth and 1,3-propanediol production.