[Feasibility and efficacy of pre-management of superior laryngeal artery in endoscopic surgery for hypopharyngeal cancer].

Objective: To explore the feasibility and efficacy for the dissection and ligation of the superior laryngeal artery in endoscopic surgery for hypopharyngeal cancer. Methods: Eight cadaveric heads were selected, and the laryngopharynxes were harvested. The positions of the superior laryngeal arteries entering the larynxes were dissected and observed under endoscopic vision, and their anatomical characteristics were summarized. Twenty-nine patients (all were male, aged 39-74 years old) with hypopharyngeal cancer who underwent transoral endoscopic surgery at the Department of Otorhinolaryngology Head and Neck Surgery of the Second Xiangya Hospital, Central South University from January 2018 to December 2019 were selected, and the patients were randomly divided into two groups by drawing lots, namely, the superior laryngeal artery was actively dissected and occluded during surgery in observation group (n=15) or not in control group (n=14). The differences in surgical time, bleeding volume, postoperative complications, and postoperative disease-free survival rate were compared between the two groups. Statistical analysis was conducted using SPSS 25.0 software. Results: The entry point of the superior laryngeal artery into the larynx was approximately at the level of the superior edge of the thyroid cartilage, and entered the larynx at the posterior one-third of the lateral wall of the pyriform fossa. The superior laryngeal artery might be determined through endoscopic exploration in all patients of observation group. The endoscopic surgery time [(40.00±7.56) minutes] and intraoperative bleeding volume [(24.00±8.28) ml] in the observation group were respectively less than those [(48.57±14.06) minutes and (42.86±15.41) ml] in the control group, and the differences were statistically significant (t=-2.064, P=0.049; t=-4.064, P=0.001). There was no case with postoperative bleeding in the observation group, but with one case of postoperative bleeding in the control group. Total disease free survival rate was 86.2% and there was no significant difference in disease free survival rates between the two groups during a follow-up period of at least 36 months (P=0.986). Conclusion: Dissection of the superior laryngeal artery during endoscopic surgery for hypopharyngeal cancer is feasible, and pre-management and occlusion of the superior laryngeal artery can effectively reduce intraoperative bleeding.

[Application of supraclavicular fasciocutaneous island flap for reconstruction after removal of tumors in parotid and auricle area].

Objective: To evaluate the efficacy of supraclavicular fasciocutaneous island flap (SIF) for repairing the defect of parotid or auricle regions after tumor resection. Methods: From February 2019 to June 2021, 12 patients (11 males and 1 female, aged 54-77 years old), of whom 4 with parotid adenoid cystic carcinoma and 8 with auricular basal cell carcinoma underwent reconstruction surgery for postoperative defects in the parotid gland area and auricular area with SIF in the Department of Otorhinolaryngology Head and Neck Surgery, the Second Xiangya Hospital of Central South University and their clinical data were retrospectively analyzed. Size of the SIF, time for harvesting SIF, neck lymph node dissection and postoperative complications were recorded. Results: The flap areas were (6-9) cm × (8-13) cm, and the harvesting time for SIF ranged from 40 to 80 min, averaging 51.7 min. The donor sites were directly closed. All patients underwent ipsilateral levels Ⅰ-Ⅲ neck dissection, with 4 cases undergoing additional level Ⅳ neck dissection and 2 cases undergoing level Ⅳ-Ⅴ neck dissection. Of the 12 SIF, 10 were completely survival and 2 had flap arterial crisis with partial flap necrosis, in addition, 1 had donor site wound dehiscence. With follow-up of 10-42 months, there were no tumor recurrences in 10 patients, 1 patient was lost to follow-up at 10 months postoperatively, and 1 patient experienced local tumor recurrence at 11 months after surgery and died 15 months later. Conclusion: SIF is an easily harvested flap with good skin features matching the skin in parotid and auricle regions and less damage to donor site, and this flap has no need for microvascular anastomosis technique. SIF is feasible and effective for repairing defects in parotid and auricle area.

[Feasibility and efficacy of preserving internal branch of superior laryngeal nerve in endoscopic surgery for hypopharyngeal squamous cancer: an observational study].

Objective: This study was performed to investigate the feasibility of preservation of internal branch of superior laryngeal nerve(ibSLN) during transoral endoscopic surgery for hypopharyngeal squamous cancer(HSCC) and the influence on patient's swallowing function after operation. Methods: From May 2020 to June 2021, the data of 29 HSCC patients who required for transoral endoscopic surgery in the Department of Otorhinolaryngology Head and Neck Surgery, the Second Xiangya Hospital of Central South University were prospectively included, and the included patients were divided into two groups randomly by lottery. According to whether ibSLN was actively dissected during operation, they were divided into ibSLN preservation group (n=15) and control group (n=14, without ibSLN preservation). Operation time, intraoperative hemorrhage, intraoperative neck dissection, postoperative radiotherapy, postoperative recurrence within 1 year, retention and swallowing function, the recovery of oral soft diet and the quality of life were compared between two groups. SPSS 25.0 software was used for statistical analysis. Results: The study included 29 eligible patients, including 25 males and 4 females.The age ranged from 42 to 67 (56.07±5.93) years. There were no significant differences(P>0.05) between 2 groups in the following data,including age(t=-0.56), gender(χ2=0.01), TNM stage(T stageχ2=0.29, N stage χ2=0.02), pathological diagnosis(χ2=0.03), preoperative swallowing function(χ2=0.00) and M. D. Anderson Dysphagia Inventory(MDADI) score(global t=0.55, emotional t=0.16, functional t=0.60, physical t=0.64), operation time(t=1.62) and intraoperative hemorrhage(t=-1.46), intraoperative neck dissection(χ2=0.01), postoperative radiotherapy(χ2=0.32), postoperative recurrence within 1 year(P>0.050). The swallowing function was evaluated by water swallowing test after operation. The swallowing function of ibSLN preservation group was better than control group, and the difference between two groups was statistically significant on the 1st (χ2=4.44, P=0.035), 5th (χ2=4.24, P=0.039) and 7th (χ2=4.55, P=0.033) day after operation. On the 14th day after operation, the MDADI scores of patients in the ibSLN preservation group were higher than those in the control group in global (t=2.45, P=0.021), functional (t=2.54, P=0.017) and physical (t=2.24, P=0.034) dimensions, except for emotional dimension (t=1.89, P=0.070). The median time of oral soft diet(U=23.00, P<0.001), normal oral diet(U=21.00, P<0.001) and the nasogastric tube removal time (U=18.50, P<0.001) in ibSLN preservation group was 2 days, 5 days and 6 days respectively, earlier than that in control group, which had statistically significant difference. Conclusion: Our results show that it is feasible to preserve the ibSLN during HSCC transoral endoscopic surgery, which can achieve rapid recovery of postoperative swallowing function.

Beclin1 enhances cisplatin-induced apoptosis via Bcl-2-modulated autophagy in laryngeal carcinoma cells Hep-2.

To investigate the role of Beclin1 in cisplatin-induced apoptosis in laryngeal carcinoma cells Hep-2 and to explore the potential mechanism. We up-regulated Beclin1 expression in Hep-2 cells. The survival rate and apoptotic rate were evaluated by MTT and flow cytometry (FCM). The Beclin1 overexpression group and the control group were treated with cisplatin for 24 hours. The proliferation and cell apoptosis of laryngeal cancer cell lines were evaluated. The mitochondrial membrane potentials were detected by DiOC6(3). Activities of Caspase-8/9/3 and convention of microtubule-associated protein one light chain 3 (LC3) were detected by western blot. The effect of Bcl-2 overexpression on increased cisplatin-sensitivity and autophagy induced by Beclin1 was investigated using Bcl-2 cDNA transfection. Expression of Beclin1 in Hep-2 cells was meaningfully enhanced by transfection, and the proliferation and the apoptosis were not considerably affected. By cisplatin treatment, the Beclin1 overexpression group showed lower survival rate and higher apoptotic rate than the control group (p<0.05). Decrease of mitochondrial membrane potential and increase of activities of Caspase-8, Caspase-9 and Caspase-3 were detected. Beclin1 overexpression increase the convention of LC3, especially after the cisplatin treatment. Overexpression of Bcl-2 decreased the cisplatin-induced apoptosis and inhibited Beclin1-induced autophagy. In conclusion, Beclin1 enhances cisplatin-induced apoptosis in laryngeal carcinoma cells Hep-2 via Bcl-2 modulated autophagy.

[Inhibition of PI3K-Akt pathway reverses LMP1 induced TRAIL resistance in nasopharyngeal carcinoma cell].

Objective:To observe the role of PI3K-Akt pathway in epsterin barr virus latent membrane protein 1(LMP1) induced TNF-related apoptosis inducing ligand(TRAIL) resistance in nasopharyngeal carcinoma(NPC) cell. Method:Expressions of Akt and p-Akt were detected after LMP1 overexpression by western blot. Colocation of LMP1 and p-Akt was detected by immunofluorescence. PI3K-Akt pathway was inhibited by LY294002 in CNE 1 and CNE 1+ LMP1. Then TRAIL sensitivity of CNE 1+ LMP1 and CNE 1 was analyzed by MTT and flow cytometric analysis, respectively. Activation of cell extrinsic apoptotic pathway was analyzed by western blot. Result:The expression of p-Akt in CNE 1+ LMP1 was higher than CNE 1. Immunofluorescence showed that after LMP1 overexpression in CNE 1, p-Akt was translocated from cytoplasm to membrane. With 1 µmol/ml LY294002 pretreatment for 8 hours, p-Akt expression was both downregulated in CNE 1 and CNE 1+ LMP1, but there was no significant difference between them(P >0.05). Moreover, the cytotoxicity and apoptosis induced by TRAIL in CNE 1 were enhanced after LMP1 overexpression and there was no significant difference between CNE 1 and CNE 1+ LMP1(P >0.05). Conclusion:LMP1 induced TRAIL-resistance in NPC cell through activeting PI3K-Akt pathway, sug-gesting that therapy targeting PI3K-Akt pathway could reverse TRAIL-resistance in NPC.

Computational characterization of substrate binding and catalysis in S-adenosylhomocysteine hydrolase.

S-Adenosylhomocysteine (AdoHcy) hydrolase catalyzes the reversible hydrolysis of AdoHcy to adenosine (Ado) and homocysteine (Hcy), playing an essential role in modulating the cellular Hcy levels and regulating activities of a host of methyltransferases in eukaryotic cells. This enzyme exists in an open conformation (active site unoccupied) and a closed conformation (active site occupied with substrate or inhibitor) [Turner, M. A., Yang, X., Yin, D., Kuczera, K., Borchardt, R. T., and Howell, P. L. (2000) Cell Biochem. Biophys. 33, 101-125]. To investigate the binding of natural substrates during catalysis, the computational docking program AutoDock (with confirming calculations using CHARMM) was used to predict the binding modes of various substrates or inhibitors with the closed and open forms of AdoHcy hydrolase. The results have revealed that the interaction between a substrate and the open form of the enzyme is nonspecific, whereas the binding of the substrate in the closed form is highly specific with the adenine moiety of a substrate as the main recognition factor. Residues Thr57, Glu59, Glu156, Gln181, Lys186, Asp190, Met351, and His35 are involved in substrate binding, which is consistent with the crystal structure. His55 in the docked model appears to participate in the elimination of water from Ado through the interaction with the 5'-OH group of Ado. In the same reaction, Asp131 removes a proton from the 4' position of the substrate after the oxidation-reduction reaction in the enzyme. To identify the residues that bind the Hcy moiety, AdoHcy was docked to the closed form of AdoHcy hydrolase. The Hcy tail is predicted to interact with His55, Cys79, Asn80, Asp131, Asp134, and Leu344 in a strained conformation, which may lower the reaction barrier and enhance the catalysis rate.

[Studies on anamorph of Cordyceps sinensis (Berk) from Naqu Tibet].

OBJECTIVE: To isolate and determine the strain of anamorph of Cordyceps sinensis collected from Naqu Tibet. METHOD: Isolating the strains of anamorph of Cordyceps sinensis collected from Baqing Shuoxian, Biru Counties by means of the ascope, tissue, ossified larva and body liquid. RESULT: Same strain of anamorph of Cordyceps sinensis was obtained with 4 different methods. CONCLUSION: The strain of Hirsutella sinensis is the anamorph of C. sinensis from Naqu Tibet.

[Investigation on quality difference between Cordyceps of Sichuan and those of Xizang].

OBJECT: The quality difference among the Cordyceps specimens which grew in various micro-ecological environment and between those from Kangding of sichuan and those from Naqu of Xizang was compared and analyzed. METHOD: The specimens from various spots in field were collected, their appearance properties were analyzed, content of adenosine was assayed, and their difference on ecological hereditary variation was discussed. RESULT: The biggest values of quality difference among the Cordyceps specimens from various micro-ecological environment can surpass that between specimen of Sichuan and specimen of Xizang. CONCLUSION: The effect of the micro-ecological environment on the quality of the Cordyceps might exceed that of extensive climates and region differences at times.

Loss of conformational stability in calmodulin upon methionine oxidation.

We have used electrospray ionization mass spectrometry (ESI-MS), circular dichroism (CD), and fluorescence spectroscopy to investigate the secondary and tertiary structural consequences that result from oxidative modification of methionine residues in wheat germ calmodulin (CaM), and prevent activation of the plasma membrane Ca-ATPase. Using ESI-MS, we have measured rates of modification and molecular mass distributions of oxidatively modified CaM species (CaMox) resulting from exposure to H2O2. From these rates, we find that oxidative modification of methionine to the corresponding methionine sulfoxide does not predispose CaM to further oxidative modification. These results indicate that methionine oxidation results in no large-scale alterations in the tertiary structure of CaMox, because the rates of oxidative modification of individual methionines are directly related to their solvent exposure. Likewise, CD measurements indicate that methionine oxidation results in little change in the apparent alpha-helical content at 28 degrees C, and only a small (0.3 +/- 0.1 kcal mol(-1)) decrease in thermal stability, suggesting the disruption of a limited number of specific noncovalent interactions. Fluorescence lifetime, anisotropy, and quenching measurements of N-(1-pyrenyl)-maleimide (PMal) covalently bound to Cys26 indicate local structural changes around PMal in the amino-terminal domain in response to oxidative modification of methionine residues in the carboxyl-terminal domain. Because the opposing globular domains remain spatially distant in both native and oxidatively modified CaM, the oxidative modification of methionines in the carboxyl-terminal domain are suggested to modify the conformation of the amino-terminal domain through alterations in the structural features involving the interdomain central helix. The structural basis for the linkage between oxidative modification and these global conformational changes is discussed in terms of possible alterations in specific noncovalent interactions that have previously been suggested to stabilize the central helix in CaM.

[Relationship between illumination and growth of the stroma of Cordyceps sinensis (Berk.) Sacc].

The growth of the stroma of Cordyceps sinensis largely depends upon the illumination in its growth period. By increasing illumination time and light intensity, its growth height can be controlled, growth rate slowed down and the corrosion time of larva body of host insect prolonged. Ultraviolet light is able to affect the growth of stroma too. The stroma also shows the strong phototaxis in its growth period. The arrangement and formation of the perithecium vary with the illumination.