PagJAZ5 regulates cambium activity through coordinately modulating cytokinin concentration and signaling in poplar.

Wood is resulted from the radial growth paced by the division and differentiation of vascular cambium cells in woody plants, and phytohormones play important roles in cambium activity. Here, we identified that PagJAZ5, a key negative regulator of jasmonate (JA) signaling, plays important roles in enhancing cambium cell division and differentiation by mediating cytokinin signaling in poplar 84K (Populus alba × Populus glandulosa). PagJAZ5 is preferentially expressed in developing phloem and cambium, weakly in developing xylem cells. Overexpression (OE) of PagJAZ5m (insensitive to JA) increased cambium activity and xylem differentiation, while jaz mutants showed opposite results. Transcriptome analyses revealed that cytokinin oxidase/dehydrogenase (CKXs) and type-A response regulators (RRs) were downregulated in PagJAZ5m OE plants. The bioactive cytokinins were significantly increased in PagJAZ5m overexpressing plants and decreased in jaz5 mutants, compared with that in 84K plants. The PagJAZ5 directly interact with PagMYC2a/b and PagWOX4b. Further, we found that the PagRR5 is regulated by PagMYC2a and PagWOX4b and involved in the regulation of xylem development. Our results showed that PagJAZ5 can increase cambium activity and promote xylem differentiation through modulating cytokinin level and type-A RR during wood formation in poplar.

PagWOX11/12a positively regulates the PagSAUR36 gene that enhances adventitious root development in poplar.

Adventitious root (AR) development is an extremely complex biological process that is affected by many intrinsic factors and extrinsic stimuli. Some WUSCHEL-related homeobox (WOX) transcription factors have been reported to play important roles in AR development, but their functional relationships with auxin signaling are poorly understood, especially the developmental plasticity of roots in response to adversity stress. Here, we identified that the WOX11/12a-SMALL AUXIN UP RNA36 (SAUR36) module mediates AR development through the auxin pathway in poplar, as well as under salt stress. PagWOX11/12a displayed inducible expression during AR development, and overexpression of PagWOX11/12a significantly promoted AR development and increased salt tolerance in poplar, whereas dominant repression of PagWOX11/12a produced the opposite phenotype. PagWOX11/12a proteins directly bind to the SAUR36 promoter to regulate SAUR36 transcription, and this binding was enhanced during salt stress. Genetic modification of PagWOX11/12a-PagSAUR36 expression revealed that the PagWOX11/12a-PagSAUR36 module is crucial for controlling AR development via the auxin pathway. Overall, our results indicate that a novel WOX11-SAUR-auxin signaling regulatory module is required for AR development in poplar. These findings provide key insights and a better understanding of the involvement of WOX11 in root developmental plasticity in saline environments.

Transcriptome Analysis of Populus euphratica under Salt Treatment and PeERF1 Gene Enhances Salt Tolerance in Transgenic Populus alba × Populus glandulosa.

Populus euphratica is mainly distributed in desert environments with dry and hot climate in summer and cold in winter. Compared with other poplars, P. euphratica is more resistant to salt stress. It is critical to investigate the transcriptome and molecular basis of salt tolerance in order to uncover stress-related genes. In this study, salt-tolerant treatment of P. euphratica resulted in an increase in osmo-regulatory substances and recovery of antioxidant enzymes. To improve the mining efficiency of candidate genes, the analysis combining both the transcriptome WGCNA and the former GWAS results was selected, and a range of key regulatory factors with salt resistance were found. The PeERF1 gene was highly connected in the turquoise modules with significant differences in salt stress traits, and the expression levels were significantly different in each treatment. For further functional verification of PeERF1, we obtained stable overexpression and dominant suppression transgenic lines by transforming into Populus alba × Populusglandulosa. The growth and physiological characteristics of the PeERF1 overexpressed plants were better than that of the wild type under salt stress. Transcriptome analysis of leaves of transgenic lines and WT revealed that highly enriched GO terms in DEGs were associated with stress responses, including abiotic stimuli responses, chemical responses, and oxidative stress responses. The result is helpful for in-depth analysis of the salt tolerance mechanism of poplar. This work provides important genes for poplar breeding with salt tolerance.

Retraction Note to: An assessment of transgenomics as a tool for gene discovery in Populus euphratica Oliv.

This article (Zhou et al. 2018) has been retracted by the authors because the sequence BIBAC 002A111F06 was incorrectly assigned to the wrong bacterial species. The BIBAC 002A111F06 sequence (GenBank Accession KC129717) reported in the paper was attributed to Populus euphratica Oliv. The BLAST search of this KC129717 sequence against the nr database at NCBI showed that it has very high similarity to a genomic sequence from the gram-negative bacteria Stenotrophomonas maltophilia. The bacterium associates with Populus euphratica Oliv. and DNA isolated from Populus euphratica Oliv. for the construction of the BIBAC clone library inlcuded DNA from Stenotrophomonas maltophilia. Therefore, the phenotype of the transgenic Arabidopsis line carrying the KC129717 sequence cannot be attributed to genes from Populus euphratica Oliv. The authors apologize for the confusion and misinterpretation of our data resulting from the incorrect sequence assignment. All authors agree to this retraction.

An assessment of transgenomics as a tool for gene discovery in Populus euphratica Oliv.

KEY MESSAGE: Transgenomics for gene discovery in Populus euphratica. Transgenomics, a member of the omics family of methodologies, is characterized as the introduction of DNA from one organism into another on a genome-wide scale followed by the identification of recipients with altered phenotypes. This strategy allows investigators to identify the gene(s) involved in these phenotypic changes. It is particularly promising for woody plants that have a long life cycle and for which molecular tools are limited. In this study, we constructed a large-insert binary bacterial artificial chromosome library of Populus euphratica, a stress-tolerant poplar species, which included 55,296 clones with average insert sizes of about 127 kb. To date, 1077 of the clones have been transformed into Arabidopsis thaliana via Agrobacterium by the floral dip method. Of these, 69 transgenic lines showed phenotypic changes represented by diverse aspects of plant form and development, 22 of which were reproducibly associated with the same phenotypic change. One of the clones conferring transgenic plants with increased salt tolerance, 002A1F06, was further analyzed and the 127,284 bp insert in this clone harbored eight genes that have been previously reported to be involved in stress resistance. This study demonstrates that transgenomics is useful in the study of functional genomics of woody plants and in the identification of novel gene(s) responsible for economically important traits. Thus, transgenomics can also be used for validation of quantitative trait loci mapped by molecular markers.

Correlation between CO2 storage at the last minute of gas insufflation and area of retroperitoneal lacuna during retroperitoneal laparoscopic radical nephrectomy.

BACKGROUND: Adequate operation interspace is the premise of laparoscopy, and carbon dioxide (CO2) was an ideal gas for forming lacuna. A retroperitoneal space is used to form operation interspace in retroperitoneal laparoscopic radical nephrectomy by making ballooning, and the retroperitoneal space has no relative complete and airtight serous membrane, therefore CO2 absorption may be greater in retroperitoneal than transperitoneal laparoscopic radical nephrectomy. Excess CO2 absorption may induce hypercapnemia and further cause physiopathological change of respiratory and circulatory system. Therefore, exact evaluation of amount of CO2 which is eliminated from body via minute ventilation is important during retroperitoneal laparoscopic radical nephrectomy. The aim of the paper is to study the correlation between CO2 storage at the last minute of gas insufflation and area of retroperitoneal lacuna during retroperitoneal laparoscopic radical nephrectomy. METHODS: Forty ASA I/II patients undergoing retroperitoneal laparoscopic radical nephrectomy were enrolled. CO2 storage at the last minute of gas insufflation and area of a retroperitoneal lacuna were observed. Linear correlation and regression were performed to determine the correlation between them. RESULTS: There was positive correlation between CO2 storage at the last minute of gas insufflation and area of retroperitoneal lacuna (r = 0.880, P = 0.000), and the equation of linear regression was y = -83.097 + 0.925x (R(2) = 0.780, t = 11.610, P = 0.000). CONCLUSIONS: Amount of CO2 which is eliminated from body via mechanical ventilation could be calculated by measuring the area of retroperitoneal lacuna during retroperitoneal laparoscopic radical nephrectomy, and an anesthetist should be aware of the size of lacuna to predict high CO2 storage at the last minute of gas insufflation.

Comparative Proteomic Analysis of Mature Pollen in Triploid and Diploid Populus deltoides.

Ploidy affects plant growth vigor and cell size, but the relative effects of pollen fertility and allergenicity between triploid and diploid have not been systematically examined. Here we performed comparative analyses of fertility, proteome, and abundances of putative allergenic proteins of pollen in triploid poplar 'ZhongHuai1' ('ZH1', triploid) and 'ZhongHuai2' ('ZH2', diploid) generated from the same parents. The mature pollen was sterile in triploid poplar 'ZH1'. By applying two-dimensional gel electrophoresis (2-DE), a total of 72 differentially expressed protein spots (DEPs) were detected in triploid poplar pollen. Among them, 24 upregulated and 43 downregulated proteins were identified in triploid poplar pollen using matrix-assisted laser desorption/ionisation coupled with time of-flight tandem mass spectrometer analysis (MALDI-TOF/TOF MS/MS). The main functions of these DEPs were related with "S-adenosylmethionine metabolism", "actin cytoskeleton organization", or "translational elongation". The infertility of triploid poplar pollen might be related to its abnormal cytoskeletal system. In addition, the abundances of previously identified 28 putative allergenic proteins were compared among three poplar varieties ('ZH1', 'ZH2', and '2KEN8'). Most putative allergenic proteins were downregulated in triploid poplar pollen. This work provides an insight into understanding the protein regulation mechanism of pollen infertility and low allergenicity in triploid poplar, and gives a clue to improving poplar polyploidy breeding and decreasing the pollen allergenicity.

[Potato Spectrum and the Digital Image Feature Parameters on the Response of the Nitrogen Level and Its Application].

In order to know the potatoes nitrogen situation rapidly and accurately, promoting the efficient use of nitrogen fertilizer on the potatoes. Using the feature of portable hyperspectral spectrometer, digital cameras and SPAD-502 chlorophyll meter to abtain the potato digital indicators, leaf spectral and SPAD. Analysing the change status of digital indicators, leaf spectral index, SPAD and production of potatoes under different nitrogen levels in two key periods. Analysing the correlation between canopy image, leaf spectral and SPAD and production, with SPAD as auxiliary validation index, nitrogen fertilizer efficiency evaluation of yield to make sure potato canopy image under the most economic nitrogen application levels and leaf spectral's critical value to explore the methods of nitrogen nutrition diagnosis quickly and simply. The results show: (1)With nitrogen levels increased, potato tuber formation stage and tuber bulking stage leaf spectral reflectance is the emergence of the "red shift" phenomenon, and the red edge parameters REP, Lwidth, FD_Max increased, Lo decreased. (2)With the nitrogen levels increased, potatoes tuber formation stage and tuber bulking stage digital indicators G/B, (G-B)/(R+G+B) decreased gradually, B/(R+G+B) increased gradually. (3) with the increase of nitrogen application rate SPAD is increased.It is obvious low nitrogen levels increase production with nitrogen increased. It is not obvious the high level of nitrogen stimulation effect. Potato canopy image, leaf spectral and red edge parameters have good correlation with SPAD value and productions, establishing the index evaluation of nitrogen nutrition abundance or lack of quantitative standard of potatoes. Indicating digital image and spectrum technology to nitrogen nutrition diagnosis of potatoes is feasible, provide research ideas and technical support for the potato accurate monitoring of nitrogen nutrition.

Portal vein clamping alone confers protection against hepatic ischemia-reperfusion injury via preserving hepatocyte function in cirrhotic rats.

BACKGROUND: Chronic liver diseases always increase the risk of liver failure after hepatectomy. We aimed to explore the protective effect of portal vein clamping without hepatic artery blood control (PVC) on a cirrhotic rat liver that underwent ischemia and reperfusion. METHODS: Carbon tetrachloride-induced cirrhotic rats were randomly assigned to four groups as follows: cirrhotic control, PVC, portal triad clamping (PTC), and intermittent portal triad clamping (IC). After 45 min of portal vascular clamping, hepatic injury and liver function were investigated by assessing the 7-d survival rate, liver blood loss, serum alanine aminotransferase, liver tissue malondialdehyde, liver tissue adenosine triphosphate, indocyanine green retention rate, and morphology changes of the rat liver. RESULTS: The 7-d survival rates in the PVC and IC groups were much higher than in the PTC group. The PVC group had more liver blood loss during the hepatectomy than the PTC group, but had much less than the cirrhotic control group (P < 0.01). In addition, there were no differences between the IC group and PVC group. The PVC rats had a significantly higher adenosine triphosphate level in the liver tissue and a markedly lower indocyanine green retention rate than the PTC and IC rats (P < 0.05). At 1, 6, and 24 h after reperfusion, the alanine aminotransferase and malondialdehyde levels in the PTC group were much higher than those in the PVC and IC groups (P < 0.05). Based on the histopathologic analysis, hepatic injury in the PVC and IC groups were similar but less prominent than in the PTC group. CONCLUSIONS: Although both PVC and IC can confer protection against hepatic ischemic-reperfusion injury in cirrhotic rats, the PVC method is more efficient in preserving the energy and function of hepatocytes than the IC method, suggesting better prognosis after hepatectomy.

The monitoring of microvascular liver blood flow changes during ischemia and reperfusion using laser speckle contrast imaging.

OBJECTIVE: The recovery of microvascular liver blood flow (LBF) after ischemia is an important determinant of the degree of hepatocellular injury. Laser speckle contrast imaging (LSCI) was recently suggested to be a suitable instrument for monitoring the LBF. This study was designed to evaluate LSCI in monitoring the LBF changes during liver ischemia and reperfusion (IR). METHODS: A rat model with 120-min ischemia and 60-min reperfusion to 90% of the liver (entire liver except the caudate lobe, which was kept as portal blood bypass) was used. The LBF of the sham operation (SO) group and the IR group was measured with LSCI at the following time points: before ischemia (Baseline), 5 min after the start of ischemia (I-5 min), 5 min before the end of ischemia (I-115 min) and 5 and 60 min after the start of reperfusion (R-5 min and R-60 min). The reproducibility among different rats or repeated measurements, the liver histopathology, the liver biological zero (BZ) and the influence of liver movement on the LSCI measurements were investigated. RESULTS: The entire exposed liver surface after laparotomy was suitable for full-view LSCI imaging. Establishing many circular or oval regions of interest (ROIs) on the LSCI flux image was a simple and convenient method for calculating and comparing the LBF of different ROIs and different liver lobes. There was good-to-moderate intra-individual and inter-individual reproducibility for the LSCI measurements of the LBF in the rats of the SO group. In the IR group, the total blood inflow occlusion resulted in a notable drop of the LBF from the baseline (P<0.05) that remained for the 120 min of ischemia. The LBF decreased further after the reperfusion (P<0.05), reflecting the IR-induced liver microcirculation dysfunction. The histopathological examination revealed severe hepatic sinus congestion and damaged hepatocytes in the IR group. The no flow BZ and liver movement contributed to the LBF values. CONCLUSIONS: LSCI technology is a simple, convenient and accurate method for the real-time monitoring of microvascular LBF changes during ischemia and reperfusion, regardless of the contribution of biological zero and liver movement. This finding suggests the possible application of LSCI for monitoring the microvascular LBF changes intraoperatively.

Computerized cognitive remediation therapy on cognitive impairment and social function in patients with chronic schizophrenia.

BACKGROUND: Patients with schizophrenia may have various disease manifestations, most of which gradually tend toward incurable chronic decline, leading to mental disability. The basic symptoms of the disease can impair social function, whereas long-term hospitalization produces hospitalization syndrome, causing serious damage to social function. AIM: To investigate the effects of Computerized Cognitive Remediation Therapy (CCRT) on cognitive and social functioning in patients with chronic schizophrenia. METHODS: A retrospective analysis of 120 patients with chronic schizophrenia in Shanghai Pudong New Area Mental Health Center was performed. They were divided into an intervention group (60 cases treated with CCRT combined with conventional medication) and a control group (60 cases treated with conventional medication). After treatment, effects on cognitive function and social roles were observed in both groups. The Positive and Negative Syndrome Scale (PANSS) was used to assess the patients' psychiatric symptoms. The Wisconsin Card Sorting Test (WCST) was used to assess the patients' cognitive functioning, and the Social Functioning Scale for Psychiatric Inpatients (SSPI) was used to assess the social functioning of the inpatient psychiatric patients. RESULTS: No significant differences were observed in the PANSS, WCST, and SSPI intergroup scores before treatment (P > 0.05). After 2, 4, and 6 wk of therapy, general psychopathological factors, positive symptoms, negative symptoms, and total PANSS scores of PANSS in the intervention group were lower than in the control group (P < 0.05). After 2, 4, and 6 wk of treatment, the number of false responses, number of persistent bugs, and total responses in the WCST were significantly lower in the intervention group than in the control group (P < 0.05), and the amount of completed classification was significantly higher than in the control group (P < 0.05). After 2, 4, and 6 wk of therapy, the SSPI scores were significantly greater than those of the controls (P < 0.05). After 6 wk of treatment, the efficacy rates of the control and intervention groups were 81.67% and 91.67%, respectively. The curative effect in the intervention group was significantly higher than that in the control group (P < 0.05). CONCLUSION: CCRT can significantly improve cognitive function and social abilities in patients with chronic schizophrenia.

Clinical Application of Ureteroscope-Assisted Laparoscopic Surgery for Prostatic Utricle in Children.

Purpose: To investigate the use of ureteroscope-assisted laparoscopic surgery (UALS) in treating symptomatic prostatic utricle (PU) in children. Materials and Methods: Data on surgically treated cases of PU at the Department of Urology in Hunan Children's Hospital between September 2014 and September 2022 were retrospectively collected and analyzed. The diagnosis was confirmed by cystourethroscopy followed by ureteroscopy, and PU was excised by ureteroscope-assisted laparoscopy. Results: A total of 21 patients with PU were enrolled in this study. The median age of the patients at surgery was 8.1 (4.6-11.5) years. Karyotyping was available for 15 children: 13 (86.7%) were 46XY, 1 (6.7%) was 45X/46XY, and 1 (6.7%) was 45X/46XY/47XYY. The median length of the PU was 5.0 (4.1-7.1) cm. Nineteen patients underwent only ureteroscope-assisted laparoscopic excision, whereas 2 also had a perineal incision. All excisions were successfully performed. The median intraoperative blood loss was 25.0 (20.0-37.5) mL. The median hospital stay and follow-up durations were 18.0 (14.5-25.0) days and 24.0 (13.5-49.0) months, respectively. The patients reported no postoperative clinical symptoms. Conclusion: UALS allows for accurate patient positioning and thorough exposure of the anatomical structures, and it is a safe, effective, and minimally invasive treatment for PU in children.

Immunoglobulin a vasculitis with testicular/epididymal involvement in children: A retrospective study of a ten-year period.

The clinical characteristics and risk factors for testicular/epididymal involvement in 73 children with immunoglobulin A vasculitis (IgAV) who were admitted to our hospital between January 2012 and November 2022 were reviewed. The demographic data, laboratory parameters, and follow-up data of the patients were compared to those of 146 males without testicular/epididymal involvement. A logistic regression analysis was performed to determine the variables associated with testicular/epididymal involvement. The prevalence of testicular/epididymal involvement among male patients with IgAV was 1.3% (73/5,556). Increased blood flow in the testes and/or epididymis on ultrasound was found in 71 patients. The remaining two patients underwent surgical exploration for loss or reduction of testicular blood flow. One patient underwent orchiectomy for intraoperative confirmation of complete right testicular infarction. Pathological findings revealed IgA immune complex deposition in the testis. Patient age (odds ratio [OR] = 0.792; 95% confidence interval [CI]: 0.682-0.919, p = 0.002), platelet count (OR = 1.011; 95% CI: 1.002-1.020, p = 0.013), and immunoglobulin M (IgM) levels (OR = 0.236; 95% CI: 0.091-0.608, p = 0.003) were strongly associated with the occurrence of testicular/epididymal involvement in IgAV. Therefore, young age, increased platelet count, and low IgM levels in patients with IgAV are potential risk factors for testicular/epididymal involvement. Doppler ultrasound can help differentiate IgAV from acute scrotum. Most patients with testicular/epididymal involvement have good prognoses, although serious complications such as testicular infarction may occur.

Allicin ameliorates cardiac hypertrophy and fibrosis through enhancing of Nrf2 antioxidant signaling pathways.

AIM: To evaluate the protective effects of allicin on Ang II-induced cardiac hypertrophy. METHODS: Sprague-Dawley male rats were randomized into 3 groups:1)sham group (saline)(n = 12), 2) Ang II group(n = 9), 3) allicin group (Ang II + allicin)(n = 9). They received infusions of either saline or Ang II (250 ng/kg body weight per min) through mini-osmotic pumps implanted subcutaneously for 2 weeks and given a diet containing 180 mg/kg/day of allicin for 8 consecutive weeks. Hemodynamic, morphological, histological, and biochemical changes were evaluated at corresponding time points. RESULTS: Ang II infusion increased blood pressure, heart rate and heart weight to body weight ratio, and resulted in anatomical and functional changes, such as increased LV mass, posterior wall thickness and LV end-diastolic diameter, and decreased fractional shortening and EF compared with sham rats. Nrf2 and HO-1 in the hearts of rats in the Ang II group were moderately elevated at both mRNA and protein levels compared to sham group mice, but NQO1 andγ-GCS were significantly lower. GPx activities, levels of GSH and T-AOC in the hearts of the rats in the Ang II group were also significantly lower, and the levels of TBARS, reactive oxygen species and protein carbonyl were significant increased. Allicin attenuated LV mass, posterior wall thickness and LV end-diastolic diameter (1.10 ± 0.04 vs. 1.37 ± 0.05, 2.26 ± 0.08 vs. 2.96 ± 0.12, 7.27 ± 0.36 vs. 8.56 ± 0.41, respectively; all P < 0.05), and increased fractional shortening and EF (28.30 ± 3.21 vs. 25.40 ± 2.57, 60.27 ± 5.63 vs. 51.30 ± 4.78, respectively; both P < 0.05) in the Ang II-induced hypertrophic rats compared to the untreated Ang II rats. Furthermore, allicin treatment attenuated the accumulation of interstitial collagen and collagen I/III (P < 0.01 vs. the untreated Ang II group), decreased the levels of reactive oxygen species, protein carbonyl and TBARS and increased GPx activities. Moreover, allicin significantly increased mRNA expression and protein levels of Nrf2, NQO1, and γ-GCS ( P < 0.01, P < 0.05 vs. the untreated Ang II group). CONCLUSION: Allicin could prevent the development of cardiac remodeling and the progression of cardiac hypertrophy to cardiac dysfunction caused by enhancing the Nrf2 antioxidant signaling pathways.

Bacillus thuringiensis Cry3Aa fused to a cellulase-binding peptide shows increased toxicity against the longhorned beetle.

Cry3 class toxins are used extensively for biological control of coleopteran larvae. We previously identified a peptide (PCx) from a phage display library that specifically binds Cx-cellulase from the midgut of Anoplophora glabripennis Motschulsky (Asian longhorn beetle) larvae. Here, we added a DNA fragment that encodes the peptide onto either end of the cry3Aa gene and tested the expressed PCx-Cry3Aa and Cry3Aa-PCx proteins for insecticidal activity in the longhorned beetle. An insect bioassay revealed that, compared with native Cry3Aa, the two modified Cry3Aa proteins had significantly higher lethality, with PCx-Cry3Aa exhibiting a mortality rate almost three times that of Cry3Aa. We also proposed that the increased lethality in larvae fed with PCx-Cry3Aa or Cry3Aa-PCx would be attributable to the binding of the toxin with Cx-cellulase, thereby increasing toxin retention in the midgut. The significantly enhanced insecticidal activity of Cry3Aa fused with the Cx-cellulase binding peptide provides a new strategy for increasing toxin efficacy against the longhorned beetle. These uniquely modified Cry3Aa proteins have potential use for pest control.

[Advance in biomechanical study of embryonic vascular system development].

Embryonic vascular system development is a complex process, whose progress is regulated by a variety of the stimulation and inhibition signals, and these signals must play synergistic effect so as to ensure that each stage of vascular development can proceed normally. The vascular development is controlled by the gene to a certain extent, and has received extensive attention. Recent studies have revealed the biomechanical role is necessary to embryonic vascular development, in which different mechanism of cell biomechanics is involved. In this review, we summarize the latest research progress on the role of biomechanical factors during embryonic vascular system development.

[Effect of pretreatment with puerarin on activation of LPS-induced RAW264. 7 cells].

OBJECTIVE: To observe the effect of pretreatment with puerarin on activation of LPS -induced RAW264. 7 cells and secretory cytokines, and discuss its anti-inflammatory mechanism. METHOD: Well-grown RAW264. 7 cells in the exponential phase were collected and randomly divided them into the blank control group, the LPS group and the puerarin pretreatment + LPS group. The cellular toxic effect of puerarin on RAW264. 7 cells was examined by CCK-8 assay, cell morphology was detected by Giemsa stain method, the changes in TNF-alpha and MIP-2 were tested by ELISA, and the expression of NF-kappaB p65 mRNA were determined by qRT-PCR. RESULTS: When puerarin was cultured with 1 mg x L(-1) LPS at a concentration of lower than 400 micromol x L(-1), it had not showed the cellular toxic effect (P < 0.05). Compared with the control group, the LPS group could significantly change the morphology of RAW264. 7 cells (increase in cell body, irregular shape, with a large number of pseudopodia extending). After intervention, the puerarin 100 micromol x L(-1) group could significantly inhibit LPS-induced cell morphological changes, while the puerarin 200 micromol x L(-1) and 400 micromol x L(-1) puerarin groups showed more notable inhibitory effects. However, there was no obvious difference between the two groups. The pretreatment with puerarin could inhibit the expression of TNF-alpha and MIP-2 in cell supernatant and NF-kappaB p65 mRNA in cells (P < 0.05). With increase in the puerarin concentration, its inhibitory effect gradually grew (P < 0.05), but did not reach the level of the blank control group. CONCLUSION: As a safe and effective natural anti-inflammatory drug, puerarin can significantly reduce the expression of inflammatory cytokines (TNF-alpha, MIP-2). Its mechanism may be related to the reduction of NF-kappaB p65 mRNA expression.

Clinical Efficacy of Laparoscopic Orchiopexy With the Modified Prentiss Maneuver for Non-palpable Testis Near the Internal Ring.

Objective: To compare the clinical efficacy and safety of laparoscopic orchiopexy with the modified Prentiss maneuver (LOMPM) and laparoscopic trans-inguinal orchiopexy (LTIO) for the treatment of non-palpable testis (NPT) <1 cm from the internal ring. Methods: Children with unilateral NPT who underwent laparoscopic orchiopexy at our center between February 2018 and January 2021 were retrospectively analyzed. According to the surgical method, they were divided into LOMPM and LTIO groups. The operation time, postoperative pain degree, postoperative complications and follow-up results were compared between the two groups. Results: A total of 98 patients were included in this study, including 41 cases in the LOMPM group and 57 cases in the LTIO group. All patients underwent successful surgery. The LOMPM group was superior to the LTIO group in terms of postoperative testicular position (lower scrotm: 90.2 vs. 71.9%, P = 0.026). There were no significant differences in operation time, postoperative pain score, and complications between the two groups. Preoperative testicular volume, postoperative testicular volume, and testicular growth rate in the LOMPM group were comparable to those in the LTIO group. There were no testicular atrophy, inguinal hernia and hydrocele in both groups after operation. Conclusions: LOMPM was comparable in safety to LTIO, but LOMPM had a good post-operative testicular position, and was suitable for the treatment of NPT near the internal ring.

Testicular regression syndrome: A retrospective analysis of clinical and histopathological features in 570 cases.

This study aimed to analyze the clinical features and pathological findings of the largest reported case series of testicular regression syndrome (TRS). Data, including age, affected side, color Doppler ultrasound results, surgical methods, intraoperative conditions, and pathological examinations, of children with unilateral TRS who were treated in our center from December 2012 to November 2021 were retrospectively analyzed. A total of 570 patients were included in this study. The mean age at surgery was 38 (range, 5-193) months. There were 457 cases (80.2%) of left TRS. Preoperative color Doppler ultrasonography found nubbins in 172 cases (30.2%). The long diameter of the contralateral testis was 17.11 (±4.22) mm, and the volume was 0.81 (±1.15) ml. The long diameter was ≥1.6 cm in 62.0% of the patients (240/387) aged ≤3 years. Laparoscopy was performed as the initial surgical step in 513 cases, of which 96.7% of the children had closed internal rings. One or more lesions of fibrosis, hemosiderin, and calcification were found in 92.4% (474/513) of the excised remnants. Germ cells were present in 16 cases (3.1%). In conclusion, TRS is more common on the left side and is usually accompanied by a closed internal ring and compensatory hypertrophy of the contralateral testis. Germ cells are only present in cases where the spermatic vessels enters the internal ring. We recommend that further exploration and excision of the remnants may not be applicable in cases where only the vas deferens has entered the internal ring.

Changes in freezing tolerance in hybrid poplar caused by up- and down-regulation of PtFAD2 gene expression.

In plant species, the level of polyunsaturated fatty acids (PUFAs) is essential for cold acclimation. To test whether changes in PUFA levels can lead to the alteration of freezing tolerance in poplar trees, we up- and down-regulated a Populus tomentosa Delta-12 fatty acid desaturase gene (PtFAD2) in the hybrid poplar (P. alba x P. glandulosa) clone 84 K. Real-time PCR results demonstrated that compared to untransformed control lines, the transcriptional level of PtFAD2 increased by up to 90% in over-expressing poplar lines (line OE-1) and decreased in down-regulated RNAi lines by up to 64% (line DR-1). As a result, the content of linoleic (C18:2) and linolenic (C18:3) unsaturated fatty acids (FAs) in total FAs increased by 7.5 and 3.9%, respectively, in the OE-1 line and decreased by 14.4 and 5.4% in the DR-2 line when compared to non-transgenic lines. After freezing treatment at -4 degrees C for 3 h without pre-cold acclimation, the survival rates of the PtFAD2-over-expressing cuttings were significantly higher (60% for OE-1) than those of non-transgenic plants (36.7%) and down-regulated lines (10% for DR-2). These results clearly demonstrate that the expression level of PUFAs substantially affected the freezing tolerance of hybrid poplar cuttings and could thus be utilized as an effective strategy to improve poplar anti-freezing traits through genetic engineering biotechnology.