New Analogues of Amides and Quinolinones Produced by Streptomyces sp. YIM S01983.

Streptomide (1), a new amide analogue, streptomynone (2), a new quinolinone, and ten known compounds including three aliphatic acids (3-5), two amides (6-7), four cyclic dipeptides (8-11), and an adenosine (12) were isolated from the fermentation broth of Streptomyces sp. YIM S01983 isolated from a sediment sample collected in Bendong Village, Huadong Town, Chuxiong, China. Their structures were determined by analysis of the 1D/2D-NMR and HR-ESI-MS spectra. Compound 12 presented weak antimicrobial activities against Candida albicans and Aligenes faecalis (MIC=64 µg/mL). Compounds 7 and 12 showed weak cytotoxic activity against MHCC97H.

Case report of avascular necrosis of the glans penis after PAE embolization.

BACKGROUND: Prostate artery embolization (PAE) is a relatively safe and effective alternative method for the treatment of lower urinary tract symptoms secondary to benign prostatic hyperplasia. The adverse events caused by PAE are primarily mild, including urinary tract infection, acute urinary retention, dysuria, fever, etc. Severe complications, such as nontarget organ embolism syndrome or penile glans ischemic necrosis, are rare. Here, we report a case of severe ischemic necrosis of the glans penis after PAE and review the literature. CASE PRESENTATION: An 86-year-old male patient was admitted to the hospital due to progressive dysuria with gross hematuria. The patient underwent placement of a three-way urinary catheter to facilitate continuous bladder flushing, hemostasis, and rehydration. After admission, his hemoglobin decreased to 89 g/L. After an examination, the diagnosis was benign prostatic hyperplasia with bleeding. During communication with the patient regarding treatment, he requested prostate artery embolization due to his advanced age and concomitant disease status. He underwent bilateral prostate artery embolization under local anesthesia. His urine gradually turned clear. However, on the 6th day after embolization, the glans gradually showed ischemic changes. On the 10th day, there was partial necrosis and blackening of the glans. The glans completely healed, and the patient was able to urinate smoothly on the 60th day after local cleaning and debridement, the administration of pain relief, anti-inflammatory and anti-infection agents, and external application of burn ointment. CONCLUSION: Penile glans ischemic necrosis after PAE is rare. The symptoms include pain, congestion, swelling, and cyanosis in the glans.

Retroperitoneal tumor finally diagnosed as a bronchogenic cyst: A case report and review of literature.

BACKGROUND: Retroperitoneal bronchogenic cyst (RBC) is an extremely rare developmental abnormality. Most are benign tumors but malignant transformation is possible. Because of their anatomical position, RBCs are easily misdiagnosed as adrenal or pancreatic solid tumors on radiological evaluation. Here, we report a case of RBC, review the literature, and summarize some important features. CASE SUMMARY: A 49-year-old woman was incidentally found to have a retroperitoneal tumor during a physical examination. Enhanced computed tomography and laboratory evaluations, including routine blood examination, blood biochemistry, 24-h urine 17 ketones, 17 hydroxyls, adrenocortical hormone, serum potassium concentration, serum amylase, lipase, and epithelial tumor markers, revealed a moderate density, 54 mm × 40 mm mass with a clear boundary near the left adrenal gland. The were no abnormalities in the blood and urine values. Because the patient had a history of hypertension and the location of the mass was adjacent to the adrenal gland, it was initially diagnosed as a left adrenal tumor and was resected by retroperitoneal laparoscopy. However, the pathological examination after surgery confirmed it to be a bronchogenic cyst. CONCLUSION: Retroperitoneal laparoscopic surgery can be prioritized for symptomatic RBC patients. Conservative treatment is feasible for selected patients.

The immuneoreaction and antioxidant status of Chinese mitten crab (Eriocheir sinensis) involve protein metabolism and the response of mTOR signaling pathway to dietary methionine levels.

To study the effects of dietary methionine on growth performance, immunity, antioxidant capacity, protein metabolism, inflammatory response and apoptosis factors in Chinese mitten crabs (Eriocheir sinensis). Five diets with different methionine levels (0.63%, 0.85%, 1.06%, 1.25% and 1.47%) were fed to E. sinensis for 8 weeks. Results showed that in the 1.25% Met group, both growth performance and feed utilization were significantly increased. The crude protein content of crab muscle in the 1.06% and 1.25% Met groups was significantly higher than that in the control group. The immune and antioxidant enzyme activities, as well as gene expression levels of anti-lipopolysaccharide factor 1 (ALF1), Crustin-1, prophenoloxidase (proPO), cap 'n' collar isoform C (CncC) in 1.25% Met group were significantly higher than other groups. The activities of adenosine deaminase (ADA) and glutamate transaminase (GPT) in serum decreased first and then increased with the increase of methionine content, while the changes of ADA and GPT in hepatopancreas increased first and then decreased. 1.25% Met group exhibited significantly increased levels of GOT, GPT, and ADA compared to the control group. 1.25% Met diet group significantly up-regulated protein synthesis and anti-apoptotic factors, and significantly down-regulated inflammatory and pro-apoptotic factors in hepatopancreas. At 1.25% in the diet, methionine was found to boost E. sinensis growth, muscle protein deposition and immunity, as well as its antioxidant capacity. Combined with the above results, based on the expression of factors involved in the mammalian target of rapamycin (mTOR) signaling pathway and the mitogen-activated protein kinase (MAPK) signaling pathway, it is proved that methionine can not only promote protein metabolism, improve feed utilization, but also alleviate the inflammatory response and apoptosis caused by oxidative stress in the body.

Construction of PD1/CD28 chimeric-switch receptor enhances anti-tumor ability of c-Met CAR-T in gastric cancer.

Chimeric antigen receptor (CAR) T cell is a promising method in cancer immunotherapy but faces many challenges in solid tumors. One of the major problems was immunosuppression caused by PD-1. In our study, the expression of c-Met in GC was analyzed from TCGA datasets, GC tissues, and cell lines. The c-Met CAR was a second-generation CAR with 4-1BB, cMet-PD1/CD28 CAR was c-Met CAR adding PD1/CD28 chimeric-switch receptor (CSR). In vitro, we measured the changes of different subgroups, phenotypes and PD-1 expression in CAR-T cells. We detected the secretion levels of different cytokines and the killing ability of CAR-Ts. In vivo, we established a xenograft GC model and observed the anti-tumor effect and off-target toxicity of different CAR-Ts. We find that the expression of c-Met was increased in GC. CD3+CD8+ T cells and CD62L+CCR7+ central memory T cells (TCM) were increased in two CAR-Ts. The stimulation of target cells could promote the expression of PD-1 in c-Met CAR-T. Compared with Mock T, the secretion of cytokines as IFN-γ, TNF-α, IL-6, IL-10 secreted by two CAR-Ts was increased, and the killing ability to c-Met positive GC cells was enhanced. The PD1/CD28 CSR could further enhance the killing ability, especially the long-term anti-tumor effect of c-Met CAR-T, and reduce the release level of IL-6. CAR-Ts target c-Met had no obvious off-target toxicity to normal organs. Thus, the PD1/CD28 CSR could further enhance the anti-tumor ability of c-Met CAR-T, and provides a promising design strategy to improve the efficacy of CAR-T in GC.

Oral Administration of Lactobacillus Casei Expressing Flagellin A Protein Confers Effective Protection against Aeromonas Veronii in Common Carp, Cyprinus Carpio.

Aeromonas veronii is a pathogen capable of infecting humans, livestock and aquatic animals, resulting in serious economic losses. In this study, two recombinant Lactobacillus casei expressing flagellin A (FlaA) of A. veronii, Lc-pPG-1-FlaA (surface-displayed) and Lc-pPG-2-FlaA (secretory) were constructed. The immune responses in fish administered with recombinant L. casei were evaluated. The two recombinant L. casei were orally administered to common carp, which stimulated high serum IgM and induced higher ACP, AKP, SOD and LYZ activity. Using qRT-PCR, the expression of IL-10, IL-8, IL-1ß, TNF-α and IFN-γ in the tissue of fish immunized with recombinant L. casei was significantly (p < 0.05) upregulated, which indicated that recombinant L. casei could activate the innate immune system to trigger the cell immune response and inflammatory response. Furthermore, recombinant L. casei was able to survive the intestinal environment and colonize in intestine mucosal. The study showed that after being challenged by A. veronii, fish administered with Lc-pPG-1-FlaA (70%) and Lc-pPG-2-FlaA (50%) had higher survival rates compared to Lc-pPG and PBS, indicating that recombinant L. casei might prevent A. veronii infection by activating the immune system to trigger immune responses. We demonstrated that flagellin as an antigen of vaccine, is acceptable for preventing A. veronii infection in fish. The recombinant L. casei expressing FlaA may be a novel mucosal vaccine for treating and controlling A. veronii.

Nobiletin inhibited hypoxia-induced epithelial-mesenchymal transition of lung cancer cells by inactivating of Notch-1 signaling and switching on miR-200b.

Epithelial-mesenchymal transition (EMT) is an early step in the process of tumor metastasis. It is well known that tumor microenvironment affects malignancy in various carcinomas; in particular, that hypoxia induces EMT. Deregulated notch signaling also contributes a lot to the development of EMT in lung cancer. In this study, we investigated the use of Notch-1-inhibiting compound as novel therapeutic candidates to regulate hypoxia-induced EMT in lung cancer cells. According to previous screening, nobiletin was selected as a Notch-1 inhibitor. Hypoxia-induced EMT was characteristic of increased N-cadherin & vimentin expressions and decreased E-cadherin expressions. Treatment with nobiletin notably attenuated hypoxia-induced EMT, invasion and migration in H1299 cells, accompanied with reduced Notch-1, Jagged1/2 expressions and its downstream genes Hey-1 and Hes-1. Nobiletin treatment also promoted tumorsuppressive miR-200b level. Moreover, notch-1 siRNA prevented hypoxia-mediated cell migration and decreased Twist1, Snail1, and ZEB1/2 expressions, which are key EMT markers. Re-expression of miR-200b blocked hypoxia-induced EMT and cell invasion. Our findings suggest that downregulation of Notch-1 and reexpression of miR-200b by nobiletin might be a novel remedy for the therapy of lung cancer.

[Auricular reconstruction for microtia with modified Nagata method].

OBJECTIVE: To investigate a method for total reconstruction of auricle. METHODS: 90 patients (101 ears) with congenital microtia underwent two-stage operations for auricular reconstruction. The first stage involved fabrication and grafting of autologous costal cartilage, removing the remnant ear cartilage, embedding the framework into local flap of the mastoid region, transferring the remnant ear lobule flap to link to the inferior framework. The second stage was creating an auriculocephalic sulcus. The reconstruction was performed 4 - 12 months after the first surgery. Skin incision was made 5 mm lateral side of the posterior margin of the auricle. The ear framework carrying a thick ear fascia was separated from the side of the head, the frames of the costal cartilage banked at the first operation were harvested, shaved and transplanted to the posterior wall of the concha with sutures; adjust stand position and angle, so that made the ear shape, position, axis, close to the healthy ear, and auriculocephalic angle was slightly larger than the contralateral ear. Two random flap was designed with superior on the root of the helix and in the inferior-posterior direction of the inferior mastoid area, two flapes were elevated and transplanted to posterior auricular sulcus to cover the grafted cartilage. Skin graft was performed in the remaining raw surface. RESULTS: A total of 90 patients were operated, all of 101 constructed ears achieved satisfied or near satisfied shapes. Five cases of partial skin flap necrosis were caused by pedicle impairment. Exposure of cartilage framework happened in two cases. The auriculocephalic sulcus of four cases diminished after the second stage operation. Three month to two-year follow-up of 67 patients showed that the reconstructed ears were satisfied with the results, including good shapes and steady auriculocephalic angles. CONCLUSIONS: The method is a simple, safe and reliable method for total aural reconstruction.

[Reconstruction of segmental ear helix defect with post-auricular skin flap].

OBJECTIVE: To explore the method of repairing segmental ear helix defect. METHODS: Twenty-one patients with segmental ear helix defect were repaired with post-auricular skin flap. In the first stage operation, ear helix defect was assessed, including the anterior and posterior area defect. According to the defect, post-auricular skin flap was designed and transplanted to repair the defect. Six weeks later, the pedicle of the post-auricular skin flap was cut off, elevated, and folded to form the helix. The secondary defect was directly sutured or repaired with skin graft. RESULTS: Twenty-one patients were treated with this method. In two to 12 months follow-up, all flaps survived and reconstructed ear helices were in good shape. The reconstructed ears were in symmetry to the healthy ones. CONCLUSION: The method is safe and effective for the correction of segmental ear helix defect.

Analysis of long-term outcomes of double-strut bone graft for osteonecrosis of the femoral head.

OBJECTIVE: To analyze the long-term effect of double-strut bone graft for osteonecrosis of the femoral head (ONFH). METHODS: A total of 366 adult patients with ONFH in 466 hips underwent double-strut bone graft from March 1988 to January 1999. Of them, 186 patients with 206 hips and an average age of 32.2 years (range, 20-60 years) were followed up for more than five years, up to January 2006. Based on the Association Research Circulation Osseous (ARCO) classification, there were 36 hips in stage IIB, and 30, 40, 40, 32 and 28 in stage IIC, IIIA, IIIB, IIIC and IV, respectively. The functional results of affected hips were evaluated by the hundred forked method. RESULTS: Hip pain in all patients disappeared or alleviated greatly after the operation. The height of the femoral head improved to various extents, and the range of motion of the hip joint increased. The patients were followed up for 5-16 years, (average 10.5 years). The total scores increased significantly postoperatively (P < 0.01). The rate of excellent and good results was 83.3%, 80.0%, 75.0%, 65.0%, 40.6% and 28.6% in stage IIB, IIC, IIIA, IIIB, IIIC and IV, respectively (63.6% for the whole group). CONCLUSION: The long-term effect of double-strut bone graft for ONFH is satisfactory in relation to staging of ONFH. Favorable results can be expected in young ONFH patients in stage IIB, IIC, and IIIA, IIIB.

The relationship between the extracellular matrix and the angiogenesis and metastasis of laryngeal carcinoma.

AIMS: We studied the relationships between the growth, metastasis and angiogenesis of laryngeal carcinoma and extracellular matrix protein-1 (ECM1) and hyaluronan (HA). METHODS: Thirty-three cases of benign and malignant laryngeal tumors were included. Using immunohistochemical staining, we examined the expression of ECM1 and HA in tumors and metastatic lymph nodes and correlated it with counts of microvessel density. RESULTS: The expression of ECM1 (p = 0.004) and HA (p = 0.036) was significantly different between benign and malignant tumors. The expression of ECM1 was strongly associated with microvessel density (Spearman's rho = 0.513, p = 0.017), with the strongest expression present within caner nests. With respect to HA, expression was significantly associated with more advanced clinical TNM stage (Spearman's rho = 0.521, p = 0.015). The link was stronger in cases where metastasis or recurrence occurred than in those without complication (p = 0.014). No significant relationship was found between ECM1 and HA expression among malignant samples (Spearman's rho = 0.383, p = 0.087). CONCLUSIONS: ECM1 has a high association with the growth, metastasis and angiogenesis of laryngeal carcinoma. An increased level of HA expression was related to the process of laryngeal carcinogenesis. Specifically, carcinoma cells with high levels of HA secretion had a higher potential for metastasis. No significant relative expression of ECM1 and HA was found.

Auricle reconstruction with a nickel-titanium shape memory alloy as the framework.

OBJECTIVE: The objective of this study is to explore the biocompatibility and implantability of a nickel-titanium (NiTi) alloy in auricle reconstruction. METHODS AND MATERIALS: Twelve New Zealand rabbits underwent subcutaneous implantation with a NiTi alloy framework shaped like the human auricle under general anesthesia. The implant was inserted after skin expansion. Implant vascularization was evaluated at months 1, 3, 6, 9, and 12 after implantation by histologic analysis. Immunohistochemical methods were used to examine expression of vascular endothelial growth factor in tissue around the implant. The fibrovascular ingrowth rate of implants was determined by bone scanning using (99m)Tc-PYP. The surface of the NiTi alloy implant was examined microscopically with scanning electron microscopy. RESULT: The implant harvested showed only partial vascularization at 1 month and completely vascularized at 3 months. The amount of vascular endothelial growth factor-positive cells was markedly increased at 6 months and reached the highest number at 3 months. The fibrovascular ingrowth rate of implant was assessed by (99m)Tc-PYP bone scan using ratios of (99m)Tc-PYP activity in placement regions versus the contralateral normal region. One rabbit had exposure of the NiTi alloy framework as a result of overlying skin flap necrosis. It was rescued with animal skin without the complete removal of the framework. All the other rabbits tolerated the implant well, and there were no complications. CONCLUSION: The NiTi alloy implant represents an alternative implant for auricular reconstruction.

[Observation of auditory brainstem response and distortion product otoacoustic emission on the animal model of autoimmune auditory neuropathy].

OBJECTIVE: To set up an animal model of autoimmune auditory neuropathy and to observe the auditory brainstem response (ABR) and distortion product otoacoustic emission (DPOAE) in guinea pigs. METHODS: The spiral ganglion and the cochlear nerve were obtained and purified by electrophoresis from 250 normal guinea pigs. The purified cochlear nerve antigen was mixed with an equal volume of complete Freunds adjuvant for immunization. Seventy guinea pigs were divided into three groups: experiment group (50 guinea pigs), control group (10 guinea pigs), normal group (10 guinea pigs). ABR, DPOAE, serum IgG levels, and morphological changes of spiral ganglion cells and the cochlear nucleus were observed. The protein expressions of the antigen were examined by immunohistochemistry and the super-structure of the auditory nerve were observed. RESULTS: The threshold of ABR response increased ranged from 10 to 25 dB in 32% (32/100 ears) of the guinea pigs. The peak latencies of waves I , III and the interpeak latency I approximately III were prolonged in the hearing loss group of guinea pigs. Prolonged peak latency of wave III was noted in hearing loss group at 2 and 3 weeks post immunization and slowly decreased to normal peak latency. The amplitude of DPOAE was no difference in the guinea pigs. The levels of serum IgG increased significantly compared with those of the control group. Inflammatory cell infiltration was observed in the cochlear nerve and the number of spiral ganglion cells detected. On the contrary, inflammatory cell infiltration was not observed in the cochlear nucleus. The cell densities and the across-sectional areas of neurons in anteroventral cochlear nucleus and posteroventral cochlear nucleus were no difference in the guinea pigs. The antigen protein distributed strictly in cochlear nerve and the spiral ganglion. Some demyelinated areas in cochlear nerve was observed in this group. The threshold of ABR response in 68% guinea pigs (68/100 ears) did not increase. The data of DPOAE and the serum IgG levels show no difference compared with the control group. There were not pathological observation in spiral ganglion cells, cochlear nucleus and cochlear nerve. CONCLUSION: An animal model of autoimmune auditory neuropathy has been set up successfully and the character of the ABR and DPOAE was observed.

[Structure and function influence of cochlear after dexamethasone intratympanic application].

OBJECTIVE: To study the changes of the structure and function of cochlear after dexamethasone intratympanic application. METHODS: Forty-five guinea pigs were divided into three groups as normal group, 0.9% sodium chloride injection group and 0.5% dexamethasone group. By using the auditory brainstem response (ABR), the superoxide dismutase (SOD) activity, the nitric oxide (NO) value and cochlear surface preparation technique, the changes of structure and function of cochlear after intratympanic Dexamethasone application has been investigated in this study. RESULTS: There was not significant difference among three groups in ABR thresholds of wave III (F = 0.5, P = 0.5). There was not significant difference among three groups in SOD activity (F = 2.45, P = 0.96). There was not significant difference among three groups in NO value (F = 3.1, P = 0.3). There was not significant difference among three groups in necrotic values of hair cells on the cochlear surface specimens in 15 mm from basal turn (F = 0.93, P = 0.45). CONCLUSION: There is no change of cochlear construction and function after intratympanic Dexamethasone application.

Functional remobilization evaluation of the paralyzed vocal cord by end-to-side neurorrhaphy in rats.

OBJECTIVE: To investigate the value of end-to-side neurorrhaphy to treat vocal cord paralysis. STUDY DESIGN: A prospective study evaluating the effects of end-to-side neurorrhaphy to treat vocal cord paralysis by means of fiberoptic laryngoscopy and nerve electromyography. METHODS: Thirty Sprague-Dawley rats were divided into experimental group 1, experimental group 2, and a control group randomly. Right recurrent laryngeal nerve (RLN) was incised, and the distal end of the RLN was anastomosed to the right phrenic nerve by end-to-side neurorrhaphy in experimental group 1 or by end-to-end nerve anastomosis in experimental group 2, respectively. The adductor nerve branch of the right RLN was incised and anastomosed to the proximal end of the right ansa cervicalis nerve by end-to-end nerve anastomosis. Fiberoptic laryngoscopy and nerve electromyography were used to examine the vocal cord movement and nerve regeneration. RESULTS: Three months after operation, this effect of end-to-side neurorrhaphy created a significant difference compared with the end-to-end nerve anastomosis (P < .05). The end-to-side neurorrhaphy did not lead to vocal cord movement compared with end-to-end nerve anastomosis. CONCLUSION: Vocal cord paralysis cannot be treated by this microsurgical technique.