Higher alcohols are one of the main by-products of Saccharomyces cerevisiae in brewing. High concentration of higher alcohols in alcoholic beverages easily causes headache, thirst and other symptoms after drinking. It is also the main reason for chronic drunkenness and difficulty in sobering up after intoxication. The main objective of this review is to present an overview of the flavor characteristics and metabolic pathways of higher alcohols as well as the application of mutagenesis breeding techniques in the regulation of higher alcohol metabolism in S. cerevisiae. In particular, we review the application of metabolic engineering technology in genetic modification of amino transferase, α-keto acid metabolism, acetate metabolism and carbon-nitrogen metabolism. Moreover, key challenges and future perspectives of realizing optimization of higher alcohols metabolism are discussed. This review is intended to provide a comprehensive understanding of metabolic regulation system of higher alcohols in S. cerevisiae and to provide insights into the rational development of the excellent industrial S. cerevisiae strains producing higher alcohols.
Alcoholic Beverages ; Alcohols/analysis* ; Fermentation ; Saccharomyces cerevisiae/metabolism* ; Saccharomyces cerevisiae Proteins/metabolism*

Objective: To preliminarily explore the value of transanal endoscopic microsurgery (TEM) in rectal cancer patients with clinical complete response (cCR) following neoadjuvant chemoradiotherapy (nCRT). Methods: Using descriptive case series method, Clinical data of 13 patients who met the criteria of nCRT and were considered to be cCR after MRI or CT scanning, digital rectal examination and colonoscopic biopsy, as well as no lymph node or distant metastasis were found, then underwent TEM from 2013 to 2016 at the Department of General Surgery of Peking Union Medical College Hospital were collected retrospectively. A 3-course combination of capecitabine and oxaliplatin (XELOX) was used for chemotherapy. Besides, a 6MV-X ray radiation was used as radiotherapy simultaneously. Six to eight weeks after completion of radiotherapy, a preoperative assessment was carried out with intrarectal ultrasound, MRI, or pelvic abdominal CT examination. TEM was performed afterwards with informed consent. Postoperative pathological findings and follow-up results were used to evaluate the value of diagnosis and treatment of TEM on those patients. Results: There were 8 males and 5 females with a median age of 63 (27-80) years. Preoperative examination showed that the lesions were located in the anterior wall in 3 cases, the posterior wall in 3 cases, the left side wall in 4 cases, and the right side wall in 3 cases. Before nCRT, the distance between tumor and anal margin was (4.8±1.1) (2.0-7.5) cm; after nCRT, this distance was (5.2±1.3) (3.0-7.5) cm. All the 13 patients underwent extended local resection of rectal cancer via TEM with the placement of urethral catheter. The average operative time was (52.2±3.7) (42-70) minutes, and the average intraoperative blood loss was (19.2±2.8) (5-30) ml. All the patients could engage in daily activities on postoperative day 1, and could cater themselves orally on postoperative day 2. The main discomfort was postoperative anal pain and foreign body sensation (n=5), which could be alleviated by non-steroidal anti-inflammatory drugs. One case had postoperative lung infection and was cured by antibiotic treatment. One case had urinary retention after removing urine catheter, and then a urine catheter was re-inserted. Average postoperative hospital stay was (2.8±2.4) (2-12) days. All specimens were completely resected via TEM. Histopathological examination confirmed that 7 specimens had achieved pathologic complete response (pCR) and the other 6 specimens had obtained partial tumor response of CAP grade 2. Seven patients with pCR received a median follow-up of 24 (8-48) months and no local recurrence or distant metastasis was reported during follow-up period. Among these 7 cases, one developed defecation dysfunction after discharge, mainly for defecation pain and even dare to defecate, who returned to normal defecation within 2 months after surgery; One developed severe anal pain within six months after surgery and the pain disappeared after symptomatic pain relief. The other 6 patients with CAP grade 2 refused to undergo further radical operation because of their strong desire in preserving anus, and received remedial adjuvant chemotherapy instead. Conclusion For rectal cancer patients with cCR after nCRT, TEM does have certain application values if the patient has a strong desire to preserve anus.

Objective To preliminarily explore the value of transanal endoscopic microsurgery (TEM) in rectal cancer patients with clinical complete response (cCR) following neoadjuvant chemoradiotherapy (nCRT). Methods Using descriptive case series method,Clinical data of 13 patients who met the criteria of nCRT and were considered to be cCR after MRI or CT scanning, digital rectal examination and colonoscopic biopsy, as well as no lymph node or distant metastasis were found, then underwent TEM from 2013 to 2016 at the Department of General Surgery of Peking Union Medical College Hospital were collected retrospectively. A 3?course combination of capecitabine and oxaliplatin (XELOX) was used for chemotherapy. Besides, a 6MV?X ray radiation was used as radiotherapy simultaneously. Six to eight weeks after completion of radiotherapy, a preoperative assessment was carried out with intrarectal ultrasound, MRI, or pelvic abdominal CT examination. TEM was performed afterwards with informed consent. Postoperative pathological findings and follow?up results were used to evaluate the value of diagnosis and treatment of TEM on those patients. Results There were 8 males and 5 females with a median age of 63 (27?80) years. Preoperative examination showed that the lesions were located in the anterior wall in 3 cases, the posterior wall in 3 cases, the left side wall in 4 cases, and the right side wall in 3 cases. Before nCRT, the distance between tumor and anal margin was (4.8±1.1) (2.0?7.5) cm; after nCRT, this distance was (5.2±1.3) (3.0?7.5) cm. All the 13 patients underwent extended local resection of rectal cancer via TEM with the placement of urethral catheter. The average operative time was (52.2±3.7) (42?70) minutes, and the average intraoperative blood loss was (19.2±2.8) (5?30) ml. All the patients could engage in daily activities on postoperative day 1, and could cater themselves orally on postoperative day 2. The main discomfort was postoperative anal pain and foreign body sensation (n=5), which could be alleviated by non?steroidal anti?inflammatory drugs. One case had postoperative lung infection and was cured by antibiotic treatment. One case had urinary retention after removing urine catheter, and then a urine catheter was re?inserted. Average postoperative hospital stay was (2.8 ± 2.4) (2?12) days. All specimens were completely resected via TEM. Histopathological examination confirmed that 7 specimens had achieved pathologic complete response (pCR) and the other 6 specimens had obtained partial tumor response of CAP grade 2. Seven patients with pCR received a median follow?up of 24 (8?48) months and no local recurrence or distant metastasis was reported during follow?up period. Among these 7 cases, one developed defecation dysfunction after discharge, mainly for defecation pain and even dare to defecate, who returned to normal defecation within 2 months after surgery; One developed severe anal pain within six months after surgery and the pain disappeared after symptomatic pain relief. The other 6 patients with CAP grade 2 refused to undergo further radical operation because of their strong desire in preserving anus, and received remedial adjuvant chemotherapy instead. Conclusion For rectal cancer patients with cCR after nCRT, TEM does have certain application values if the patient has a strong desire to preserve anus.

Objective To preliminarily explore the value of transanal endoscopic microsurgery (TEM) in rectal cancer patients with clinical complete response (cCR) following neoadjuvant chemoradiotherapy (nCRT). Methods Using descriptive case series method,Clinical data of 13 patients who met the criteria of nCRT and were considered to be cCR after MRI or CT scanning, digital rectal examination and colonoscopic biopsy, as well as no lymph node or distant metastasis were found, then underwent TEM from 2013 to 2016 at the Department of General Surgery of Peking Union Medical College Hospital were collected retrospectively. A 3?course combination of capecitabine and oxaliplatin (XELOX) was used for chemotherapy. Besides, a 6MV?X ray radiation was used as radiotherapy simultaneously. Six to eight weeks after completion of radiotherapy, a preoperative assessment was carried out with intrarectal ultrasound, MRI, or pelvic abdominal CT examination. TEM was performed afterwards with informed consent. Postoperative pathological findings and follow?up results were used to evaluate the value of diagnosis and treatment of TEM on those patients. Results There were 8 males and 5 females with a median age of 63 (27?80) years. Preoperative examination showed that the lesions were located in the anterior wall in 3 cases, the posterior wall in 3 cases, the left side wall in 4 cases, and the right side wall in 3 cases. Before nCRT, the distance between tumor and anal margin was (4.8±1.1) (2.0?7.5) cm; after nCRT, this distance was (5.2±1.3) (3.0?7.5) cm. All the 13 patients underwent extended local resection of rectal cancer via TEM with the placement of urethral catheter. The average operative time was (52.2±3.7) (42?70) minutes, and the average intraoperative blood loss was (19.2±2.8) (5?30) ml. All the patients could engage in daily activities on postoperative day 1, and could cater themselves orally on postoperative day 2. The main discomfort was postoperative anal pain and foreign body sensation (n=5), which could be alleviated by non?steroidal anti?inflammatory drugs. One case had postoperative lung infection and was cured by antibiotic treatment. One case had urinary retention after removing urine catheter, and then a urine catheter was re?inserted. Average postoperative hospital stay was (2.8 ± 2.4) (2?12) days. All specimens were completely resected via TEM. Histopathological examination confirmed that 7 specimens had achieved pathologic complete response (pCR) and the other 6 specimens had obtained partial tumor response of CAP grade 2. Seven patients with pCR received a median follow?up of 24 (8?48) months and no local recurrence or distant metastasis was reported during follow?up period. Among these 7 cases, one developed defecation dysfunction after discharge, mainly for defecation pain and even dare to defecate, who returned to normal defecation within 2 months after surgery; One developed severe anal pain within six months after surgery and the pain disappeared after symptomatic pain relief. The other 6 patients with CAP grade 2 refused to undergo further radical operation because of their strong desire in preserving anus, and received remedial adjuvant chemotherapy instead. Conclusion For rectal cancer patients with cCR after nCRT, TEM does have certain application values if the patient has a strong desire to preserve anus.

BACKGROUNDIntegrin-linked kinase (ILK) is a Ser/Thr protein kinase. Many studies have showed that ILK was closely related to occurrence, proliferation, invasion and metastasis in many malignant tumors, and it appeared to be an upstream cross point of tumor-associated factors. The aim of this study is to explore the relationship between the expression of ILK and some clinical pathological factors in human non-small cell lung cancer (NSCLC), and analyze whether there is relativity between ILK and E-cadherin.

METHODSImmunohistochemical S-P method was adopted to detect the expression of ILK and E-cadherin proteins in 76 NSCLC cases with the neighboring noncancerous tissue, and the expressions of them in 30 fresh NSCLC samples were determined with Western Blot assay.

RESULTSImmunohistochemically, the overexpression of ILK protein in NSCLC was 53/76 (69.7%), including 33/44 (75.0%) squamous cell carcinoma and 20/32 (62.5%) adenocarcinoma, but its expression was not related to the histological type (P= 0.247 ). Expression of ILK was related to differentiation (rs=-0.296, P=0.009), lymph node metastasis (rs=0.311, P=0.006) and clinical stage (rs=0.350, P=0.002). Moreover, Kaplan-Meier survival estimates showed a significant correlation between ILK expression and patient survival in Log-rank test (P=0.006). Overexpression of ILK in NSCLC was associated with unfavorable prognosis. An inverse correlation between the levels of ILK and E-cadherin was found (rs=-0.514, P < 0.001). Western Blot result showed that the level of ILK in the tumor tissues was noticeably higher than that in the normal tissues (t=-6.811, P=0.0002), and an inverse correlation between the levels of ILK and E-cadherin was proved (P=0.001).

CONCLUSIONSIn NSCLC, ILK can interact with some tumor-associated factors, through which it appears to be involved in several oncogenesis-related events ,including promotion of cell survival ,as well as cell migration and invasion.ILK keeps significant inverse correlation to E-cadherin,andit would be one of the pathways for ILK to affect differentia-tion,clinical stage ,lymph node metastasis and prognosis of patients .ILK expression can be a useful predictorof poor prognosis in NSCLC,and the detections of ILK and E-cadherin will help us better to predict prognosisof patients .

Objective To investigate the spectrum of mitochondrial DNA (mtDNA) mutations in Chinese patients with Leber′s hereditary optic neuropathy (LHON). Methods The primary mtDNA mutations (G3460A?G11778A and T14484C) of 140 patients with LHON were detected by mutation-specific priming polymerase chain reaction (MSP-PCR), heteroduplex-single strand conformation polymorphism polymerase chain reaction (HA-SSCP), restriction fragment length polymorphisms (RFLP) and measurement of DNA sequence. The transmissibility of the patients′ stirps was analyzed. Results In the 140 patients with LHON, G11778A mtDNA primary mutation was found in 130 (92.9%), including 113 males and 17 females; G3460A mutation was found in 2 (1.4%) including 1 male and 1 female; G14484A mutation was found in 8 (5.7%) including 6 males and 2 females. Conclusion In Chinese patients with LHON, the incidence of G11778A mtDNA mutation is higher than that of G3460A and T14484C.

Objective　To apply two-dimensional (2-D) gel electrophoresis to resolve specially expressed proteins related to retinitis pigmentosa (RP) in the retina of rds mice.Methods　Proteins, prepared from the retinas of rds mice and normal C3B mice at different ages, were separated by using two-dimensional electrophoresis and then analyzed by 2-DE imaging analyzer.Results　2-D gel electrophoresis was established for the retinal proteome analysis. Retinal neuronal tissue was lysed by using chemical lysis solution and ultrasonic. Using carrier ampholyte to set up pH gradient as first dimension and casting vertical 12% SDS-acylamide-bis slab as second dimension, the major retinal proteins showed maps of proteome on 2-D gels clearly. Retinal proteome of rds and C3B mice at 37d has different expressive patterns. Some proteins only expressed in the retina of rds mice while another only in the retina of C3B mice and others had different level between the two kinds of mice.Conclusion　2-D gel electrophoresis is effective to separate specially expressed retinal proteins. The expressed proteins in the rds retina are different in quality and quantity from that in the normal C3B mice.

Objective　 To clone the differentially expressed genes in the retina of rds mouse (the animal model of congenital retinitis pigmentosa) during the disease development. Methods　 The retinal mRNA of rds mouse during the development of retinitis pigmentosa was analyzed by the mRNA differential display. The differentilly expressed mRNA fragments were cloned and sequenced. Results There was obvious difference of gene expression between rds mouse and the control during the development of retinitis pigmentosa. Five differentially expressed bands were cloned and sequenced. One of those had 86% identity (132/154) with the sequence of the human cDNA DKFZp434D1227 from adult testis in GenBank, which was submitted lately (15-Oct-1999) and without much information. The other had lower identity with the sequeces in GenBank. A highly expressed clone in the rds mouse on postnatal day 25 had the same length as another clone in the normal on postnatal day 37, which was not expressed in the rds mouse on day 37. The sequences of the two clones were identical in all but two base pairs. Conclusion　 These results indicate that there are a lot of novel differentially expressed genes in the chronic processing diseases, such as retinitis pigmentosa.

Objective　To determine the sensitivity and specificity of using the computer-photoscreener and non-cycloplegic retinoscopy in the detection of amblyopiogenic factors in nine to fifty months old infants.Methods　Three hundred children whose ages range from nine to fifty months were screened with the computer-photoscreener and non-cycloplegic retinoscopy. With a masked standardized clinical assessment as the standard, an overall comparison of the results obtained with the two techniques revealed a sensitivity and specificity. Photoscreen images on the computer monitor screen were reviewed and analyzed immediately by two independent observers for indicators of amblyopiogenic risk factors. Simultaneously, the results were compared to the findings of a full ophthalmologic examination.Results　The computer-photoscreener revealed a sensitivity of 94.2% and specificity of 90.1%, and the non-cycloplegic retinocopy revealed a sensitivity of 85.7% and specificity of 81.1% for the detection of amblyopiogenic risk factors, including hyperopia (+2.75 D or more), myopia (-1.50 D or more), astigmatism (2.00 D or more),anisometropia (2.00 D or more), ocular misalignment (5 degrees or more), and media opacity (1.5mm or more). Conclusion　The computer-photoscreener offers an opportunity to identify problems that limit vision, and could provide a feasible and sufficiently reliable screening technique in infants and preschool children who can be screened successfully for amblyopiogenic risk factors.

Objective To analyze the clinical characteristics and to screen for causative mutations in CRX and GUCY2D genes in children with cone or cone-rod dystrophy. Methods Clinical data and genomic DNA was collected from 18 children with cone or cone-rod dystrophy, aged from 4 months to 8 years. The coding sequence of the cone-rod homeobox (CRX) gene and two exons of the retinal-specific guanylate cyclase GUCY2D gene (exons 2 and 8) were analyzed by using polymerase chain reaction(PCR) and heteroduplex combined with single-strand conformational polymorphism (heteroduplex-SSCP) analysis. Results All of the 18 patients manifested obvious visual impairment. Nystagmus, photophobia and mild ocular fundus changes were found in 13, 8,and 7 cases respectively. Normal fundus was seen in 11 cases. The visual acuity was less than 0.3 in 4 cases and was unable to measure in the other 14 cases because they were too young. Clinical ocular manifestations between cone and cone-rod dystrophy were overlapped. Mutation in the CRX and GUCY2D genes was not detected in the 18 children with cone and cone-rod dystrophy. Conclusion Visual impairment appeared more early and obvious than fundus changes in children with cone or cone-rod dystrophy. Mutation in the CRX gene may not contribute to this series of patients with cone and cone-rod dystrophy.