Metabolomic and morphologic surveillance reveals the impact of lactic acid-treated barley on in vitro ruminal fermentation.

Objective: Lactic acid (LA) treatment of cereals is known to improve ruminant performance. However, changes in cereal nutrient levels and variations in rumen fermentation remain unclear. Method: This study was designed to compare the effects of 5% LA treatment on the trophic and morphological characteristics of barley and to discover the differences in rumen fermentation characteristics and metabolomes between LA-treated and untreated barley. Results: Compared with those of untreated barley (BA), the dry matter (DM), crude protein (CP), ash and water-soluble carbohydrate contents of barley plants treated with 5% LA for 48 h (BALA) decreased, but the resistant starch (RS) and non-fiber carbohydrate contents increased. Moreover, the amount of proteinaceous matrix in BA decreased in response to LA treatment. During in vitro fermentation, BALA had a greater pH but lower dry matter disappearance and ammonia, methane, and short-chain fatty acid levels than BA. The differential metabolites between BA and BALA were clustered into metabolic pathways such as purine metabolism, lysine degradation, and linoleic acid metabolism. Observable differences in ultrastructure between BALA and BA were noted during fermentation. Conclusion: Lactic treatment altered barley nutrient content, including DM, CP, RS, ash, water-soluble carbohydrates and non-fiber carbohydrates, and affected barley ultrastructure. These variations led to significant and incubation time-dependent changes in the in vitro fermentation characteristics and metabolome.

Condensed tannin-induced variations in the rumen metabolome and the correlation with fermentation characteristics in goats.

In this study, we characterized the effects of CT dietary inclusion at 2% (wt/wt) dry matter on the goat rumen metabolome and fermentation characteristics. Barley (BA) and corn (CN) were separately used as basal grain for the control rations, and rations supplemented with CT were BACT and CNCT, respectively. The rations were tested using eight Japanese Shiba × Saanen goats in a replicated 4 × 4 Latin square arrangement (28 days for each period). Ruminal fluid was obtained on day 25 of each period, and ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) analysis was performed. Metabolites from BACT against BA and CNCT against CN were mostly associated with purine metabolism. Moreover, BACT against BA showed intensified biosynthesis of unsaturated fatty acids, and CNCT against CN resulted in strengthened amino acid metabolism. Furthermore, strong correlations were observed between rumen NH3 -N and the copy number of total bacteria with most of the differential metabolites. The present paper provides a better understanding of the relationship between the rumen metabolome and fermentation characteristics and supports a shift in concern about using CT as a strategy to manipulate rumen metabolism.

Treatment of corn with lactic acid delayed in vitro ruminal degradation without compromising fermentation: a biological and morphological monitoring study.

Grain processed by lactic acid (LA) is known to improve ruminant growth and health. However, the exact mechanism regarding rumen hydrolysis of LA-treated grain is still ambiguous. This experiment was designed to compare the effects of 5% LA treatment on the trophic and morphological variations in corn and to discover the alternations in ruminal hydrolysis between LA-treated and untreated corn macroscopically and microscopically using in vitro fermentation method. The results showed that, compared with untreated corn (CN), corn treated with 5% LA for 48 h (CNLA) experienced a decrease in the dry matter, albumin fraction, aNDFom, and water-soluble carbohydrate content but an increase in the resistant starch content. The in vitro fermentation showed that the pH of CNLA was higher, but dry matter disappearance was lower than that of CN. Most of the fermentation indices were unaffected, except for decreased iso-butyrate and iso-valerate. The abundances of total bacteria, Prevotella spp., Streptococcus bovis, and Selenomonas ruminantium were higher, but those of Ruminococcus flavefaciens and Ruminococcus albus were lower in CNLA than in CN. There were differences in the scanning electron micrographs between CNLA and CN after 3 h of fermentation. This study suggests that treating corn with LA for 48 h can induce changes in its nutrient composition and alter the bacterial flora during subsequent in vitro fermentation. These changes appeared to be crucial contributors to the beneficial effects observed in rumen fermentation.

[Clinical application effect of pedunculated rectus abdominis muscle combined with bilateral ureters for repairing refractory bladder-vaginal stump fistula through external vesical drainage].

Objective: To investigate the efficacy and safety of pedunculated rectus abdominis combined with bilateral ureteral extravestheter drainage in the treatment of refractory bladder-vaginal stump fistula. Methods: The clinical data of 8 cases of the refractory bladder-vaginal stump fistula were admitted to the Second Hospital of Hebei Medical University and Henan Cancer Hospital and underwent the clinical treatment of bladder-vaginal stump from December 2019 to December 2022 were collected. The reason of refractory bladder-vaginal stump fistula was analyzed, the operation manner of pedunculated rectus abdominis combined with peduncle and bilateral ureter for the treatment of bladder-vaginal stump through extrabladder drainage was explored. The operation time, bleeding volume and clinical effect were record. Results: The median operation time of 8 patients was 150 minutes(120~180 min), and the median blood loss was 400 ml(200~600 ml). During the perioperative period, there were 2 cases of incision infection, delayed healing by debridement and dressing, 2 cases of incision rupture and suture wound healing after reoperation, and 2 cases of urinary tract infection were cured by anti-infection. When followed up for 6 months, 8 cases of vesicovaginal stump fistula were cured. Conclusion: Bilateral ureteral external drainage of the rectus abdominis muscle, has a practical effect in the treatment of refractory bladder-vaginal stump fistula, which can be one of the clinical repairing treatment.

Comparison of clinical outcomes between posterolateral approach and modified Hardinge approach in salvage total hip arthroplasty after medial buttress plate failure for femoral neck fractures.

OBJECTIVE: The purpose of this study was to investigate the clinical outcomes of salvage total hip arthroplasty (THA) after medial buttress plate surgery for femoral neck fractures via the modified Hardinge approach (MHA) and posterolateral approach (PLA) through a retrospective analysis. PATIENTS AND METHODS: From October 2016 to October 2020, a total of 41 patients with failed femoral neck fractures treated with cannulated screws and medial buttress plates underwent unilateral salvage THA, and a retrospective study was conducted. According to the surgical approach, patients were divided into PLA group and MHA group. Clinical and radiological data were evaluated. The primary outcome indicators were the Pain Visual Analog Scale (VAS) and Hip Harris Score (HHS). Secondary outcome indicators include hemoglobin (HGB), hematocrit (HCT), creatine kinase (CK), creatine kinase-MB (CK-MB), etc. The occurrence of postoperative complications was also recorded. RESULTS: There were no differences in demographic or clinical characteristics before surgery. There were no differences in postoperative HGB, HCT, CK-MB and radiological parameters. The surgical approach had no effect on the hospitalization period. The PLA group had earlier ambulation time, and the serum level of CK was also low. Analysis of the HHS and VAS showed that on postoperative day 3, the PLA group had superior scores. The incidence of complications did not significantly differ between groups. CONCLUSIONS: The posterolateral approach for salvage THA provides better functional recovery with less muscle damage in the early postoperative period.

Subcentimetre thyroid nodules: Sonographic features associated with malignancy.

INTRODUCTION: The aim of this study was to identify the ultrasound features which are associated with malignancy in subcentimetre thyroid nodules. METHODS: This retrospective study included 454 thyroid nodules <10 mm in size in 413 patients from 2012 to 2016, which were subjected to fine needle aspiration cytology. Each nodule was classified according to the ultrasound patterns of the 2015 American Thyroid Association guidelines and the high suspicion ultrasound features (solid, hypo-echogenicity, irregular margins, microcalcifications, taller-than-wide, interrupted rim calcifications, and extrathyroidal extension) were identified for evaluation of their diagnostic performance. RESULTS: Of the American Thyroid Association high suspicion ultrasound features, univariate analysis showed that hypo-echogenicity (sensitivity 81.6% (95% CI 65.7-92.3%), specificity 50.0% (95% CI 43.4-56.6%)), irregular margins (sensitivity 34.2% (95% CI 19.6-51.4%), specificity 92.2% (95% CI 88.0-95.3%)), microcalcifications (sensitivity 23.7% (95% CI 11.4-40.2%), specificity 91.0% (95% CI 86.5-94.3%)), and taller-than-wide (sensitivity 23.7% (95% CI 11.4-40.2%), specificity 92.2% (95% CI 88.0-95.3%)) were significantly associated with a malignant diagnosis. Amongst the above features, subsequent multivariate analysis identified a combination of hypo-echogenicity and irregular margins as significantly associated with malignancy. Our malignancy rates based on American Thyroid Association ultrasound patterns from benign to high suspicion were 0.0, 8.3, 3.9, 15.7, and 40.4%, respectively. The American Thyroid Association high suspicion ultrasound pattern had a sensitivity of 50.0% (95% CI 33.4-66.7%) and specificity of 84.5% (95% CI 79.2-88.9%). CONCLUSION: The presence of both hypo-echogenicity and irregular margins was significantly associated with malignancy in subcentimetre thyroid nodules in our study. Hence, subcentimetre nodules which are hypoechoic with irregular margins may warrant further follow-up.

Effects of the acid-base treatment of corn on rumen fermentation and microbiota, inflammatory response and growth performance in beef cattle fed high-concentrate diet.

Beef cattle are often fed high-concentrate diet (HCD) to achieve high growth rate. However, HCD feeding is strongly associated with metabolic disorders. Mild acid treatment of grains in HCD with 1% hydrochloric acid (HA) followed by neutralization with sodium bicarbonate (SB) might modify rumen fermentation patterns and microbiota, thereby decreasing the negative effects of HCD. This study was thus aimed to investigate the effects of treatment of corn with 1% HA and subsequent neutralization with SB on rumen fermentation and microbiota, inflammatory response and growth performance in beef cattle fed HCD. Eighteen beef cattle were randomly allocated to three groups and each group was fed different diets: low-concentrate diet (LCD) (concentrate : forage = 40 : 60), HCD (concentrate : forage = 60 : 40) or HCD based on treated corn (HCDT) with the same concentrate to forage ratio as the HCD. The corn in the HCDT was steeped in 1% HA (wt/wt) for 48 h and neutralized with SB after HA treatment. The animal trial lasted for 42 days with an adaptation period of 7 days. At the end of the trial, rumen fluid samples were collected for measuring ruminal pH values, short-chain fatty acids, endotoxin (or lipopolysaccharide, LPS) and bacterial microbiota. Plasma samples were collected at the end of the trial to determine the concentrations of plasma LPS, proinflammatory cytokines and acute phase proteins (APPs). The results showed that compared with the LCD, feeding the HCD had better growth performance due to a shift in the ruminal fermentation pattern from acetate towards propionate, butyrate and valerate. However, the HCD decreased ruminal pH and increased ruminal LPS release and the concentrations of plasma proinflammatory cytokines and APPs. Furthermore, feeding the HCD reduced bacterial richness and diversity in the rumen. Treatment of corn increased resistant starch (RS) content. Compared with the HCD, feeding the HCDT reduced ruminal LPS and improved ruminal bacterial microbiota, resulting in decreased inflammation and improved growth performance. In conclusion, although the HCD had better growth performance than the LCD, feeding the HCD promoted the pH reduction and the LPS release in the rumen, disturbed the ruminal bacterial stability and increased inflammatory response. Treatment of corn with HA in combination with subsequent SB neutralization increased the RS content and helped counter the negative effects of feeding HCD to beef steers.

[Analysis of predictors for Frey syndrome after parotidectomy].

Objective:To analysis the data of the patients with parotid tumors, clarify the contributing factors of Frey syndrome, and to evaluate the role of soft tissues membrane SIS in prevention of Frey syndrome after parotidectomy. Method:The data of 95 patients who suffered from parotid tumors and underwent parotidectomy were included in this study. The relationship between the patients' age, sex, tumor location ,tumor size, disease pathology, type of resection, SIS application and the incidence of Frey syndrome were statistically analyzed . Result:The incidence of Frey syndrome after parotidectomy for 95 patients was 25.3%. Age, sex, tumor location and size, disease pathology, type of resection did not appear to be associated with development of Frey syndrome（P>0.05）. SIS application was the only statistically significant factor（P<0.01）, and SIS could prevent Frey syndrome after parotidectomy. Conclusion:Frey syndrome is one of the common complicationsafter parotidectomy. Implantation of SIS is an effective method for prevention of Frey syndrome after parotidectomy.

[Primary small cell neuroendocrine carcinoma of the tonsil: a case report].

Neuroendocrine carcinoma(NEC) of the tonsil is extremely rare that exhibits highly malignant and poorly prognosis. Only a few of case reports and the optial treatment is still uncertain. Here, the current study presents a case of 60-year-old male with neck mass rapidly growing for four months. It was diagnosed a primary tonsil neuroendocrine carcinoma. And he was treated by one cycle of cisplatin and etoposide. It had no effect and the tumor was rapidly progressed. With review of the literatures, the nomenclature, clinicopathological characteristics and treatment modalities of this rare tumor are discussed.

Aberrant expression of Rab1A and its prognostic significance in human colorectal cancer.

OBJECTIVE: Colorectal cancer (CRC) is one of the most common types of cancer worldwide. Emerging evidence has verified that Rab1A plays an oncogenic role in several human malignancies including breast cancer, lung cancer, and hepatocellular carcinomas. However, the clinical significance and prognostic impact of Rab1A in CRC is still unclear. PATIENTS AND METHODS: We initiated our investigation by immunohistochemistry and Western blot analysis to confirm Rab1A expression in CRC tissues. Meanwhile, the correlation of Rab1A expression and clinicopathologic features, as well as outcome in CRC patients, were retrospectively analyzed. RESULTS: In the issue, Rab1A is overexpressed in CRC tissues compared with matched noncancerous tissues. Meanwhile, high Rab1A expression was significantly associated with the TNM stage, lymph node metastasis, and peritoneal metastasis. In addition, multivariate analyses identified Rab1A expression and TNM stage as independent predictors for CRC patients. Moreover, Kaplan-Meier survival analysis showed that patients with high Rab1A expression had a significantly worse survival time than those with low Rab1A expression, which especially affected the survival in CRC patients with advanced stage. Spearman analysis suggested that there was a positive relationship between Rab1A expression and preoperative serum carcinoembryonic antigen (CEA) for CRC patients. CONCLUSIONS: These results suggested that Rab1A is an important diagnostic marker for CRC, and Rab1A can be used as a valuable biomarker for prognosis as well as peritoneal metastasis in CRC patients. Rab1A may prove to be clinically useful for developing a new therapeutic target of CRC treatment.

Prokaryotic functional expression and activity comparison of three CYP9A genes from the polyphagous pest Helicoverpa armigera.

Cytochrome P450s (CYPs or P450s) have been long recognized as very important enzymes in the metabolism of xenobiotic and endogenous compounds, but only a few CYPs have been functionally characterized in insects. The effort in functional characterization of insect P450s is heavily hindered by technical difficulties in preparing active, individual P450 enzymes directly from the target insect. In this paper, we describe the functional expression of two additional pyrethroid resistance-associated CYP9A genes (CYP9A12 and CYP9A17) from the polyphagous pest Helicoverpa armigera in the facile Escherichia coli. The functionality of E. coli produced CYP9A12, CYP9A14, and CYP9A17 was investigated and activities of these CYP9As were compared against three probe substrates after reconstitution with NADPH-dependent cytochrome P450 reductase. The results showed that active forms of CYP9A12 and CYP9A17 were expressed in E. coli with a content of about 1.0-1.5 nmol mg-1 protein in membrane preparations. In vitro assays showed that CYP9A14 was capable of catalyzing O-dealkylation of methoxyresorufin (MROD), ethoxyresorufin (EROD), and benzyloxyresorufin (BROD), while CYP9A12 and CYP9A17 exhibited only MROD and EROD activities. Kinetic studies demonstrated that CYP9A14 had the greatest k cat/K m value for MROD, and CYP9A17 for EROD, while the lowest k cat/K m values for both MROD and EROD were observed for CYP9A12. The distinct biochemical traits suggest that the three paralogous CYP9As may play different roles in xenobiotic metabolism in this important pest.

Expression and clinical significance of ARTN and MMP-9 in endometrial carcinoma.

The aim of this study was to detect the differential expression of Artemin (ARTN) and matrix metallopeptidase protein 9 (MMP-9) in endometrial carcinoma (EC) and to assess their clinical significance in order to provide insight into the pathological mechanism of tumor infiltration and metastasis in EC. A total of 48 patients who had undergone surgery for EC at the School of Medicine and Affiliated Hospital of HeBei University of Engineering between July 2015 and July 2016 were included in the study. The 48 patients were classified into 3 groups according to tumor stage: 27 patients with EC stage I, 12 patients with EC stage II and 9 patients with EC stage III. The samples collected from each patient included fresh normal endometrial tissue, endometrial simple hyperplastic tissue and endometrial atypical hyperplastic tissue. The transcription levels of ARTN and MMP-9 mRNA in each group were investigated using RT-PCR. The expression levels of ARTN and MMP-9 protein in each group were examined using Western blotting. Spearman’s correlation analysis was used to analyze the correlation between the expression levels of ARTN and MMP-9 proteins and EC tissue type. RT-PCR and Western blotting assays revealed that the expression levels of ARTN and MMP-9 were increased in normal endometrial tissue, simple hyperplastic tissue, atypical hyperplastic tissue and EC of stages I, II and III. The differences noted were statistically significant (P less than 0.05). Furthermore, Western blot analysis indicated that the expression levels of ARTN and MMP-9 proteins in lymphatic metastatic tissues were higher than those in non-lymphatic metastatic tissues (P less than 0.05). The expression levels in the infiltration tissues of the deep muscular layer were higher than those noted in the light muscular layer (P less than 0.05). The expression levels of ARTN and MMP-9 proteins were positively correlated (P less than 0.05). The data suggest that ARTN and MMP-9 are involved in the occurrence, development, invasion and metastasis of EC, and play a synergistic role in the development of EC and lymphatic metastasis.

MR Imaging Grading System for Skull Base Chordoma.

BACKGROUND AND PURPOSE: Skull base chordoma has been widely studied in recent years, however, imaging characteristics of this tumor have not been well elaborated. The purpose of this study was to establish an MR imaging grading system for skull base chordoma. MATERIALS AND METHODS: In this study, 156 patients with skull base chordomas were retrospectively assessed. Tumor-to-pons signal intensity ratios were calculated from pretreatment MR images RT1 (ratio of tumor to pons signal intensity in T1 FLAIR sequence), RT2 (ratio of tumor to pons signal intensity in T2 sequence) and REN (ratio of tumor to pons signal intensity in enhanced T1 FLAIR sequence), and significant ratios for overall survival and progression-free survival were selected to establish a grading system. Clinical variables among different MR imaging grades were then analyzed to evaluate the usefulness of the grading system. RESULTS: RT2 (P < .001) and REN (P = .04) were identified as significant variables affecting progression-free survival. After analysis, the classification criteria were set as follows: MR grade I, RT2 > 2.49 and REN ≤ 0.77; MR grade II, RT2 > 2.49 and REN > 0.77, or RT2 ≤ 2.49 and REN ≤ 0.77; and MR grade III, RT2 ≤ 2.49 and REN > 0.77. MR grade III tumors had a more abundant tumor blood supply than MR grade I tumors (P < .001), and the intraoperative blood loss of MR grade III tumors was higher than that of MR grade I tumors (P = .002). Additionally, skull base chordoma progression risk increased by 2.071 times for every single MR grade increase (P < .001). CONCLUSIONS: A higher RT2 value was a negative indicator of tumor progression, whereas a higher REN value was a positive risk factor of tumor progression. MR grade III tumors showed a more abundant blood supply than MR grade I tumors, and the risk of skull base chordoma progression increased with every single MR grade increase.

Comparative study of 13 candidate genes applying multi-reference normalization to detect the expression of different fineness in skin tissues of wool sheep.

Fiber diameter is a useful indicator of wool traits and it is the main determinant of wool quality and value. A comparative study was conducted to analyze the abundance and expression of 13 candidate genes using expression profile microarray analysis and to identify novel molecular markers associated with wool traits to provide a molecular basis for improving wool quality in sheep. Genes associated with fineness of skin tissue were identified using a real-time reverse transcriptase-polymerase chain reaction method with 18SrRNA, ß-Actin, and GAPDH used for multi-reference normalization. The results indicated that the expression levels of TXNIP, TFDP1, and FAIM genes in super-fine type wool sheep were higher than those in fine-type wool sheep; the corresponding expression ratios of super-fine to fine wool sheep were 1.45, 1.57, and 2.55, respectively. The expression levels of PIK3CA, ADAM9, and FZD3 genes were lower in super-fine wool sheep compared with fine-type wool sheep; the corresponding expression ratios were 0.61, 0.65, and 0.52, respectively. The other genes tested (RPS6KA, ABCG2, GSTA1, PTPN13, GJB3, PPARD, and LAMB1) were similarly expressed in both types of wool sheep. These results infer that lower expression of PIK3CA, ADAM9, and FZD3 genes was associated with lower fiber diameter, whereas lower expression of TXNIP, TFDP1, and FAIM genes was associated with higher fiber diameter.

Identification and expression patterns of Halloween genes encoding cytochrome P450s involved in ecdysteroid biosynthesis in the cotton bollworm Helicoverpa armigera.

20-Hydroxyecdysone (20E) is a key hormone which regulates growth, development and reproduction in insects. Although cytochrome P450 enzymes (P450s) participating in the ecdysteroid biosynthesis of 20E have been characterized in a few model insects, no work has been published on the molecular entity of their orthologs in the cotton bollworm Helicoverpa armigera, a major pest insect in agriculture worldwide. In this study, four cytochrome P450 homologs, namely HarmCYP302A1, HarmCYP306A1, HarmCYP314A1 and HarmCYP315A1 from H. armigera, were identified and evolutional conservation of these Halloween genes were revealed among lepidopteran. Expression analyses showed that HarmCYP302A1 and HarmCYP315A1 were predominantly expressed in larval prothoracic glands, whereas this predominance was not always observed for HarmCYP306A1 and CYP314A1. The expression patterns of Halloween genes indicate that the fat bodies may play an important role in the conversion of ecdysone into 20E in larval-larval molt and in larval-pupal metamorphosis, and raise the possibility that HarmCYP315A1 plays a role in tissue-specific regulation in the steroid biosynthesis in H. armigera. These findings represent the first identification and expression characterization of four steriodogenic P450 genes and provide the groundwork for future functional and evolutionary study of steroid biosynthesis in this agriculturally important pest.

Effect of different alcohols on stratum corneum kallikrein 5 and phospholipase A2 together with epidermal keratinocytes and skin irritation.

OBJECTIVES: The aim of this exploratory study was to investigate the effect of ethanol, isopropanol and n-propanol on stratum corneum (SC) enzymes and keratinocytes in vitro together with their effects on skin condition and function. METHODS: Activities of kallikrein 5 (KLK5) and phospholipase A2 (PLA2) as well as keratinocyte metabolic activity, interleukin-1α (IL-1α) and tumor necrosis factor-α (TNF-α) were measured in vitro in the presence and absence of the different alcohols. We also measured transepidermal water loss (TEWL), skin capacitance, visual dryness and visual redness on the volar forearms of 25 Caucasian women following application of the alcohols 20 and 100 times per day over a period of 14 days in a clinical study. RESULTS: Reduced activities of KLK5 and PLA2 were observed in the presence of the alcohols. The greatest denaturing effect was always observed for n-propanol (P < 0.001), and in the case of PLA2, the effect of isopropanol was greater than ethanol (P < 0.001). Equally, ethanol had the mildest effects on keratinocyte metabolic activity and cytokine secretion (P < 0.001) and n-propanol always produced the most severe changes in normal and differentiated keratinocytes. These in vitro findings supported the clinical results where the major effects were on the induction of skin irritation (increased dropout rates) and ranked the intolerance of the different alcohols as follows: n-propanol > isopropanol > ethanol. At the high application frequencies, the effect of the different alcohols on transepidermal water loss (TEWL) and skin capacitance was similar, but at the low application frequencies, n-propanol had a significant effect on TEWL and capacitance values (P < 0.05). Equally, n-propanol and isopropanol produced significantly more skin redness at the low application frequencies. CONCLUSIONS: Clearly, isopropanol and n-propanol caused significant SC and keratinocyte perturbation in vitro together with damage to skin condition and function in vivo whereas ethanol did not. As a result, we show that ethanol-based sanitizers are better tolerated by skin, particularly in high-use settings, than other alcohols and should be the active ingredient of choice.