Rhizosphere soil properties of waxy sorghum under different row ratio configurations in waxy sorghum-soybean intercropping systems.

To overcome the continuous planting obstacle and promote the sustainable production of waxy sorghum, a two-years field experiment was performed to determine the responses of waxy sorghum rhizosphere soil properties to different row ratio configurations in waxy sorghum-soybean intercropping systems. The treatments included five row ratio configurations, which were two rows of waxy sorghum intercropped with one row of soybean (2W1S), two rows of waxy sorghum intercropped with two rows of soybean (2W2S), three rows of waxy sorghum intercropped with one row of soybean (3W1S), three rows of waxy sorghum intercropped with two rows of soybean (3W2S), and three rows of waxy sorghum intercropped with three rows of soybean (3W3S), and sole cropping waxy sorghum (SW) was used as control. The nutrients, enzyme activities, and microbes of waxy sorghum rhizosphere soil were investigated at the jointing, anthesis, and maturity stages. Results showed that rhizosphere soil properties of waxy sorghum were significantly affected by row ratio configurations of waxy sorghum intercropped soybean. Among all treatments, the performances of rhizosphere soil nutrients contents, enzymes activities, and microbes contents were 2W1S > 3W1S > 3W2S > 3W3S > 2W2S > SW. Compared to SW treatment, the 2W1S treatment increased the organic matter, total N, total P, total K, gram-negative bacteria phospholipid fatty acids (PLFAs), and gram-positive bacteria PLFAs contents and catalase, polyphenol oxidase, and urease activities by 20.86%-25.67%, 34.33%-70.05%, 23.98%-33.83%, 44.12%-81.86%, 74.87%-194.32%, 81.59-136.59%, 91.44%-114.07%, 85.35%-146.91%, and 36.32%-63.94%, respectively. Likewise, the available N, available P, available K, total PLFAs, fungus PLFAs, actinomycetes PLFAs, and bacteria PLFAs contents under the 2W1S treatment were 1.53-2.41, 1.32-1.89, 1.82-2.05, 1.96-2.91, 3.59-4.44, 9.11-12.56, and 1.81-2.71 times than those of SW treatment, respectively. Further, the determining factors of soil microbes were total K, catalase, and polyphenol oxidase for total microbes, bacteria, and gram-negative bacteria, total P and available K for fungus, available N, available K, and polyphenol oxidase for actinomycetes, and total K and polyphenol oxidase for gram-positive bacteria. In conclusion, the 2W1S treatment was the optimal row ratio configuration of waxy sorghum intercropped with soybean, which can improve the rhizosphere soil quality and promote the sustainable production of waxy sorghum.

Expression and clinicopathological characteristics of PDX1, PTF1A, and SALL4 in large and small ducts of ectopic pancreas located in gastro-duodenum and jejunum.

An ectopic pancreas is defined as pancreatic tissue outside its normal location, anatomically separated from the pancreas. The transcription factor pancreas/duodenum homeobox protein 1 (PDX1) is involved in maintaining the pancreas and functions in early pancreatic development, beta cell differentiation, and endocrine non beta cells. Pancreatic transcription factor 1 subunit alpha (PTF1A) affects exocrine cell formation and regulation of acinar cell identity, and is expressed in exocrine cells as a transcription factor. The depletion of SALL4 disrupts self-renewal and induces differentiation. To clarify which of PDX1, PTF1A, or SALL4 determines the difference in Heinrich's classification, we examined the localization and number of positive cells. We analyzed the differential expression of PDX1, PTF1A, and SALL4 in large and small ducts in ectopic pancreas by immunohistochemistry. Results showed that the number of PTF1A-positive cells in large ducts was more widespread in type I than in type II in the gastro-duodenum, and more SALL4-positive cells were noticed in large ducts than in small ducts in the gastro-duodenum of type II. Our results revealed that PTF1A might promote exocrine differentiation in developing the pancreatic tissues, and that those with widespread expression differentiate into exocrine cells.

Tunneling nanotubes-based intercellular mitochondrial trafficking as a novel therapeutic target in dry eye.

Cell-to-cell mitochondria transfer via tunneling nanotubes (TNTs) has recently been revealed as a spontaneous way to protect damaged cells. Previously, we have reported mesenchymal stem cells (MSCs) can rescue retinal ganglion cell and corneal epithelium through intercellular mitochondrial trafficking. Mitochondrial damage and oxidative stress in corneal epithelial cells are vital in dry eye disease (DED). However, whether intercellular mitochondrial transfer is involved in the pathological and repair process of DED is currently unknown. Therefore, in this study, we designed a coculture system to evaluate the role of intercellular mitochondrial transfer between human corneal epithelial cells (CEC) in DED. In addition, we successfully discovered the ROCK inhibitor, Y-27632 as an intensifier to improve the efficiency of intercellular mitochondrial transport. As expected, the enhanced mitochondrial transfer promotes the regeneration of CECs. Moreover, through further exploration of mechanisms, it was demonstrated that F-actin-mediated cell morphological changes and cytoskeletal remodeling may be potential mechanisms for Y-27632 to induce mitochondrial metastasis. In conclusion, we established a new method for cell repair in DED that healthy CEC offered mitochondria to damaged CEC, providing a new insight into the cellular mechanism of corneal epithelium homeostatic regenerative therapeutics in DED.

Effect of Corneal Collagen Cross-Linking on Subsequent Corneal Fungal Infection in Rats.

Purpose: The purpose of this study was to determine whether corneal collagen cross-linking (CXL) alters fungal susceptibility and increases the severity of keratitis through macrophage activation in rats. Methods: Four weeks following CXL pretreatment, the corneal epithelium of adult rats was removed and inoculated with Candida albicans (C. albicans; CXL + inoculation group). The non-CXL-pretreated corneas were also inoculated with C. albicans (inoculation group). Clinical scoring and histopathological examination were performed to determine the severity of fungal keratitis. Immunofluorescence and confocal microscopy imaging were applied to determine the effects of CXL treatment on corneal local macrophage content. Real-time polymerase chain reaction (RT-PCR) and Western blots were used to evaluate mRNA and protein expression. Flow cytometry assays were performed to detect M1- and M2-type macrophages. Results: CXL pretreatment (CXL + inoculation) resulted in higher infection success rate and more severe fungal keratitis than inoculation alone (inoculation group). On days 1, 3, and 7 following fungal infection, the increase in macrophage infiltration and IL-1ß, MMP-9, and VEGFA expression was greater in the CXL + inoculation group than in the inoculation group. Number of M1- and M2-type macrophages, M1 to M2 ratio, M1-type macrophage genes, inducible nitric oxide synthase (iNOS), and tumor necrosis factor (TNFα) expression were higher in the CXL + inoculation group compared with the inoculation group. Conclusions: Our data demonstrate that CXL may increase the colonization of macrophages and activate more M1-type macrophages to increase fungal susceptibility and severity of keratitis. Translational Relevance: This study may aid long-term risk assessment and treatment of the complications of CXL.

Anti-inflammatory and immunomodulatory effects of polysaccharide extracted from Wuguchong (maggot) on 2,4-dinitrochlorobenzene-induced atopic dermatitis in mice.

Atopic dermatitis (AD) is an inflammatory, heterogeneous, chronic skin disorder characterized by recurrent eczematous lesions and intense pruritus, and the pathophysiology mechanism of AD is known for immune dysregulation and inflammatory responses. Wuguchong (maggot) has been widely used in the wound field and found with pharmacological properties of the anti-inflammatory and immunomodulatory function. Recently, some polysaccharides were proven to have beneficial effects on AD skin lesions in mice and humans. However, the effect of the polysaccharide extracted from Wuguchong (PEW) on AD remains to be investigated. In the present study, we examined the anti-inflammatory and immunomodulatory effects of PEW on AD and explored the potential mechanisms. Balb/c mice were orally administrated with PEW to evaluate the therapeutic effect of PEW on 2,4-dinitrochlorobenzene (DNCB)-induced AD. Oral PEW administration significantly ameliorated the lesions and symptoms in AD mice, such as the ear thickness and ear swelling degree, epidermal and dermal thickness, and the infiltration of mast cells. In addition, PEW treatment decreased the levels of serum IgE and histamine, the frequencies of Th1 and Th17 cells, as well as the mRNA expression levels of Th1 and Th17 cytokines and nuclear transcript factors (IFN-γ, T-bet, IL-17A, and ROR-rt). Furthermore, the activation of the NF-κB pathway and the phosphorylation of MAPKs (p38, ERK, and JNK) were significantly suppressed by PEW treatment. Taken together, our study suggests that PEW exerts anti-inflammatory and immunomodulatory effects through inhibition of Th1 and Th17 responses and downregulation of NF-κB and MAPK pathways, PEW would be developed as a promising immune therapy for AD.

AMPK/MFF Activation: Role in Mitochondrial Fission and Mitophagy in Dry Eye.

Purpose: To assess the role of mitochondrial morphology and adenosine monophosphate-activated protein kinase (AMPK)/mitochondrial fission factor (MFF) in dry eye and the underlying mechanisms. Methods: Immortalized human corneal epithelial cells (HCECs) and primary HCECs were cultured under high osmotic pressure (HOP). C57BL/6 female mice were injected subcutaneously with scopolamine. Quantitative real-time PCR was used to measure mRNA expression. Protein expression was assessed by western blot and immunofluorescence staining. Mitochondrial morphology was observed by confocal microscopy and transmission electron microscopy. Results: First, HOP induced mitochondrial oxidative damage to HCECs, accompanied by mitochondrial fission and increased mitophagy. Then, AMPK/MFF pathway proteins were increased consequent to HOP-induced energy metabolism dysfunction. Interestingly, the AMPK pathway promoted mitochondrial fission and mitophagy by increasing the recruitment of dynamin-related protein 1 (DRP1) to the mitochondrial outer membrane in the HOP group. Moreover, AMPK knockdown attenuated mitochondrial fission and mitophagy due to HOP in HCECs. AMPK activation triggered mitochondrial fission and mitophagy. Mitochondrial fission of HCECs stressed by HOP was mediated via MFF phosphorylation. MFF knockdown reversed mitochondrial fragmentation and mitophagy in HCECs treated with HOP. Inhibition of MFF protected HCECs against oxidative damage, cell death, and inflammation in the presence of HOP. Finally, we detected mitochondrial fission and AMPK pathway activation in vivo. Conclusions: The AMPK/MFF pathway mediates the development of dry eye by positively regulating mitochondrial fission and mitophagy. Inhibition of mitochondrial fission can alleviate oxidative damage and inflammation in dry eye and may provide experimental evidence for treating dry eye.

Changes in Nutrient Accumulation and Transportation of Waxy Sorghum in Waxy Sorghum-Soybean Intercropping Systems Under Different Row Ratio Configurations.

To determine the optimal row ratio configuration of waxy sorghum-soybean intercropping systems, a field experiment with seven treatments, including sole waxy sorghum (SW), sole soybean (SS), two rows of waxy sorghum alternated with one row of soybean (2W1S), two rows of waxy sorghum alternated with two rows of soybean (2W2S), three rows of waxy sorghum alternated with one row of soybean (3W1S), three rows of waxy sorghum alternated with two rows of soybean (3W2S), and three rows of waxy sorghum alternated with three rows of soybean (3W3S), was conducted during 2019 and 2020 in Guiyang, China. Accumulation and transportation of nitrogen (N), phosphorus (P), and potassium (K) in waxy sorghum were investigated. The results showed that the row ratio configurations had significant effects on the N, P, and K accumulation and transportation of waxy sorghum. On the one hand, compared to SW treatment, intercropping treatments showed higher N, P, and K contents and accumulation amounts, N, P, and K transportation amounts before anthesis, N, P, and K transportation rates before anthesis, and contribution rates of N, P, and K transportation before anthesis to the grain of each organ in waxy sorghum. Similarly, the waxy sorghum-soybean intercropping system increased the yield components (including spike length, grain number per spike, and 1,000-grain weight) of waxy sorghum. In addition, the yields of waxy sorghum and soybean among all treatments were in the sequence of SW (SS) > 2W1S > 3W1S > 3W2S > 3W3S > 2W2S. Besides, the 2W1S treatment showed the highest land equivalent ratio and economic benefit. On the whole, the waxy sorghum-soybean intercropping system can increase the N, P, and K absorption among organs and promote the N, P, and K transportation from vegetative organs to grain in waxy sorghum so as to promote the growth and development of spike in waxy sorghum to obtain higher land equivalent ratio and economic benefits. The 2W1S treatment was recommended as the optimal row ratio configuration of the waxy sorghum-soybean system to achieve the maximum utilization of nutrient resources.

Protective effect of homogeneous polysaccharides of Wuguchong (HPW) on intestinal mucositis induced by 5-fluorouracil in mice.

BACKGROUND: In hospitalized patients, drug side effects usually trigger intestinal mucositis (IM), which in turn damages intestinal absorption and reduces the efficacy of treatment. It has been discovered that natural polysaccharides can relieve IM. In this study, we extracted and purified homogenous polysaccharides of Wuguchong (HPW), a traditional Chinese medicine, and explored the protective effect of HPW on 5-fluorouracil (5-FU)-induced IM. METHODS AND RESULTS: First, we identified the physical and chemical properties of the extracted homogeneous polysaccharides. The molecular weight of HPW was 616 kDa, and it was composed of 14 monosaccharides. Then, a model of small IM induced by 5-FU (50 mg/kg) was established in mice to explore the effect and mechanism of HPW. The results showed that HPW effectively increased histological indicators such as villus height, crypt depth and goblet cell count. Moreover, HPW relieved intestinal barrier indicators such as D-Lac and diamine oxidase (DAO). Subsequently, western blotting was used to measure the expression of Claudin-1, Occludin, proliferating cell nuclear antigen, and inflammatory proteins such as NF-κB (P65), tumour necrosis factor-α (TNF-α), and COX-2. The results also indicated that HPW could reduce inflammation and protect the barrier at the molecular level. Finally, we investigated the influence of HPW on the levels of short-chain fatty acids, a metabolite of intestinal flora, in the faeces of mice. CONCLUSIONS: HPW, which is a bioactive polysaccharide derived from insects, has protective effects on the intestinal mucosa, can relieve intestinal inflammation caused by drug side effects, and deserves further development and research.

CST6 protein and peptides inhibit breast cancer bone metastasis by suppressing CTSB activity and osteoclastogenesis.

Background: Bone metastasis is a frequent symptom of breast cancer and current targeted therapy has limited efficacy. Osteoclasts play critical roles to drive osteolysis and metastatic outgrowth of tumor cells in bone. Previously we identified CST6 as a secretory protein significantly downregulated in bone-metastatic breast cancer cells. Functional analysis showed that CST6 suppresses breast-to-bone metastasis in animal models. However, the functional mechanism and therapeutic potential of CST6 in bone metastasis is unknown. Methods: Using in vitro osteoclastogenesis and in vivo metastasis assays, we studied the effect and mechanism of extracellular CST6 protein in suppressing osteoclastic niches and bone metastasis of breast cancer. A number of peptides containing the functional domain of CST6 were screened to inhibit bone metastasis. The efficacy, stability and toxicity of CST6 recombinant protein and peptides were evaluated in preclinical metastasis models. Results: We show here that CST6 inhibits osteolytic bone metastasis by inhibiting osteoclastogenesis. Cancer cell-derived CST6 enters osteoclasts by endocytosis and suppresses the cysteine protease CTSB, leading to up-regulation of the CTSB hydrolytic substrate SPHK1. SPHK1 suppresses osteoclast maturation by inhibiting the RANKL-induced p38 activation. Importantly, recombinant CST6 protein effectively suppresses bone metastasis in vitro and in vivo. We further identified several peptides mimicking the function of CST6 to suppress cancer cell-induced osteoclastogenesis and bone metastasis. Pre-clinical analyses of CTS6 recombinant protein and peptides demonstrated their potentials in treatment of breast cancer bone metastasis. Conclusion: These findings reveal the CST6-CTSB-SPHK1 signaling axis in osteoclast differentiation and provide a promising approach to treat bone diseases with CST6-based peptides.

Micelle-solubilized axitinib for ocular administration in anti-neovascularization.

The development of new blood vessels is directly related to the occurrence of eye diseases. Anti-angiogenic drugs can theoretically be extended to the treatment of ophthalmic diseases. In this study, axitinib, a class of tyrosine kinase inhibitors, was loaded via the amphiphilic copolymer MPEG-PCL, improving its dispersibility in water. Axitinib-loaded micelles showed low toxicity in concentration gradient assays. Additionally, multiple doses by scratch assay confirmed that axitinib had no significant effect on normal cell migration, and biosafety test results showed good cell compatibility. After we established the corneal neovascularization model after an alkali burn in rats, the anti-angiogenic efficacy was tested, with dexamethasone as a positive control. The results showed that axitinib-loaded micelles had anti-angiogenic effects without obvious tissue toxicity. As a class of targeted tyrosine kinase inhibitors, axitinib can be used in the treatment of ocular neovascular diseases through nanocrystallization.

Differential effects on lung and bone metastasis of breast cancer by Wnt signalling inhibitor DKK1.

Metastatic cancer is a systemic disease, and metastasis determinants might elicit completely different effects in various target organs. Here we show that tumour-secreted DKK1 is a serological marker of breast cancer metastasis organotropism and inhibits lung metastasis. DKK1 suppresses PTGS2-induced macrophage and neutrophil recruitment in lung metastases by antagonizing cancer cell non-canonical WNT/PCP-RAC1-JNK signalling. In the lungs, DKK1 also inhibits WNT/Ca2+-CaMKII-NF-κB signalling and suppresses LTBP1-mediated TGF-ß secretion of cancer cells. In contrast, DKK1 promotes breast-to-bone metastasis by regulating canonical WNT signalling of osteoblasts. Importantly, targeting canonical WNT may not be beneficial to treatment of metastatic cancer, while combinatory therapy against JNK and TGF-ß signalling effectively prevents metastasis to both the lungs and bone. Thus, DKK1 represents a class of Janus-faced molecules with dichotomous roles in organotropic metastasis, and our data provide a rationale for new anti-metastasis approaches.

Combination of dexamethasone and Avastin® by supramolecular hydrogel attenuates the inflammatory corneal neovascularization in rat alkali burn model.

Corneal neovascularization (CNV) is one of the leading causes of vision loss and a high-risk factor for transplant rejection. The present study proposed a supramolecular hydrogel comprised of MPEG-PCL micelles and α-cyclodextrin (α-CD) for co-delivery of dexamethasone sodium phosphate (Dexp) and Avastin® (Ava), and further evaluated its therapeutic efficacy in rat alkali burn model. A physical mixing of Dexp/Ava, MPEG-PCL micelles, and α-CD aqueous solution leads to a spontaneous formation of the supramolecular hydrogel via a "host-guest" recognition between MPEG and α-CD. The supramolecular hydrogel provides a relatively quick release of Dexp over Ava during the study of the 5-day in vitro release. The results of in vitro cytotoxicity test and wound healing assay illustrated that the proposed supramolecular hydrogel was non-toxic against L-929 and HCEC cells and did not significantly affect the migration of HCEC cells after 24h incubation. The corneal distribution test suggested that the precorneal duration of Ava was significantly extended by the supramolecular hydrogel with respect to its solution formulation. Moreover, the supramolecular hydrogel showed high ocular biocompatibility and was a non-irritant after topical instillation. Furthermore, the Dexp-Ava hydrogel medication, but not by Ava solution and Ava hydrogel medication, could greatly attenuate the alkali burn-induced corneal inflammation and remarkably suppress the corneal neovascularization via the downregulation of VEGF, CD31, and α-SMA expression in the rat alkali burn model. As a result, the combined Dexp and Ava by supramolecular hydrogel exhibited an advantage over Ava monotherapy approach, which might be a promising alternative therapy for inflammatory CNV.

Combined sildenafil with vacuum erection device therapy in the management of diabetic men with erectile dysfunction after failure of first-line sildenafil monotherapy.

OBJECTIVE: To evaluate the efficacy and safety of combination therapy of sildenafil plus vacuum erection devices in men with type 2 diabetes mellitus with moderate to severe erectile dysfunction who are dissatisfied with the results of using sildenafil alone. METHODS: The study included 66 diabetes mellitus patients presenting erectile dysfunction for at least 6 months and dissatisfied with the use of 100 mg sildenafil monotherapy. The patients were randomized in two groups. Those in group A (n = 33) were instructed to use a vacuum erection device only, whereas those in group B (n = 33) were treated with combination therapy, including sildenafil 100 mg and a vacuum erection device. Erectile function was evaluated subjectively using the International Index of Erectile Function, Sexual Encounter Profile questionnaire questions 2 and 3 at visit 1 (baseline; study entry), visit 2 (4 weeks after baseline), and visit 3 (12 weeks after baseline; study end). RESULTS: There were no significant differences in average patient age, duration of diabetes, duration of erectile dysfunction, baseline International Index of Erectile Function scores, hypertension, blood testosterone, smoking and alcohol consumption between two groups. Mean International Index of Erectile Function scores were significantly higher for group B at the 1-month (14.86 ± 2.17 vs 12.41 ± 2.63; P < 0.0001) and 3-months (17.53 ± 2.95 vs 14.29 ± 2.81; P < 0.0001) visits. Men in group B had better successful penetration (73.3% vs 46.6%) and successful intercourse (70% vs 46.6%) at 3 months compared with group A. CONCLUSION: Combined use of sildenafil and vacuum erection device therapy significantly enhances erectile function, and it is well tolerated by diabetes mellitus patients not responding to first-line sildenafil alone.

Simultaneous saline irrigation during retrograde rigid ureteroscopic lasertripsy for the prevention of proximal calculus migration.

OBJECTIVE: In this paper, we analyze the clinical efficacy of a simultaneous saline irrigation method in treating upper-mid ureteral stone migration and evaluate its effectiveness during ureteroscopic lasertripsy. METHODS: We prospectively evaluated 78 patients with a total of 95 upper-mid ureteral stones, which were treated with holmium:YAG lasertripsy. These patients were randomized into 2 groups. In Group 1 (39 cases with 44 ureteral stones), conventional ureteroscopic lasertripsy was performed. In Group 2, (39 cases with 51 ureteral stones), the simultaneous saline irrigation method was used during lasertripsy. There was no significant difference between the groups with regards to stone site, size or state of the upper urinary tract by spiral computed tomography or excretory urography. Data were analyzed regarding stone migration, lengths of time, and ureteral clearing for various stages of each procedure. RESULTS: One patient in Group 2 (2%) experienced upward stone migration, while this occurred in 8 patients in Group 1 (20%). The operative time in Group 1 ranged from 35 to 55 minutes (mean: 44.8 ± 5.3), while in Group 2 it ranged from 40 to 69 minutes (mean: 50.4±3). There was no significant different in the operative times between the two groups (p < 0.05). Ureteral perforation, urinoma and urosepsis were not seen in both groups. CONCLUSION: The simultaneous saline irrigation method demonstrated a statistically significant advantage over conventional methods. The operation can be performed persistently under clear vision, and since the stones cannot move upward, fragmented portions are easily flushed out. Our data suggest that this method is simple, safe and effective in preventing proximal stone migration during ureteroscopic lasertripsy.

Treatment of ipsilateral renal ureteral calculi by combining retroperitoneal laparoscopic surgery with tubeless mini-percutaneous nephrolithotomy.

OBJECTIVES: To report and discuss the role of the treatment of ipsilateral renal and upper middle ureteral stones in patients by combing retroperitoneal laparoscopic surgery with tubeless mini-percutaneous nephrolithotomy. PATIENTS AND METHODS: Eleven patients associated with ipsilateral renal and upper middle ureteral stones underwent combing retroperitoneal laparoscopic surgery with tubeless mini-percutaneous nephrolithotomy after failure of shockwave or ureteroscopy lithotripsy. Their data were analyzed retrospectively including stone burden, perioperative complications and outcomes. RESULTS: All the patients underwent retroperitoneal laparoscopic ureterolithotomy and tubeless mini-percutaneous nephrolithotomy successfully. Surgical time ranged from 80 to 160 min with a mean of 118 min. The mean hospital stay was 4 days (3-7 days). The mean length of retroperitoneal urinary drainage was 3 days (2-6 days). There were minor complications in 3 (27.2%) patients. The stone-free rate was 82% (9 patients). Two patients had a residual calyceal fragment that was treated with shockwave lithotripsy. All of them were followed up for 3-24 months. Renal function was improved in different degree. CONCLUSION: In carefully selected patients, combining retroperitoneal laparoscopic surgery with tubeless mini-percutaneous nephrolithotomy can treat ipsilateral renal and upper middle ureteral calculi by a single procedure with advantages of high stone-free rate, safety, reliability, rapid recovery and less complications.