[Related factors and prognosis analysis of esophagorespiratory fistula after esophageal cancer surgery].

Objective: To explore the factors associated with the development of esophagorespiratory fistula (ERF) after esophageal cancer surgery and its relationship with patient survival. Methods: A total of 241 patients with esophageal cancer after surgery, who received postoperative sputum suction through bronchoscope from West China Hospital of Sichuan University between January and December 2021 were included. The clinical data and airway features under bronchoscope of these patients were collected. Of the 241 patients, 203 were males (84.2%) and 38 were females (15.8%), aged (63.63±8.05) years. The related factors of ERF were analyzed by multivariate logistic regression analysis, and Kaplan-meier was used to analyze the relationship between bronchoscopic specific manifestations, treatment modality and patient survival. Results: Of the 241 postoperative patients with esophageal cancer, 21 (8.7%) developed ERF. There were 39 (16.2%) patients with bronchoscopic specific manifestations, including 16 cases (6.6%) of hyperemia, 13 cases (5.4%) of congestion, and 15 cases (6.2%) of erosion. Bronchoscopic specific manifestations of tracheal mucosa (OR=13.734, 95%CI: 3.535-29.074, P<0.001) and thoracotomy (OR=9.121, 95%CI 1.843-44.237, P=0.007) were independent risk factors for the development of ERF, and preoperative chemotherapy (OR=0.128, 95%CI: 0.052-0.607, P=0.006) was a protective factor in the occurrence of ERF. The median survival time was 224 (95%CI: 95-353)d in the stent-treated group (14 patients) after the onset of ERF, and the median survival time of patients in the supportive care group (7 patients) was 29 (95%CI: 8-50)d, and the survival difference was statistically significant (χ2=5.69, P=0.017). Conclusions: Bronchoscopic specific manifestations are independent risk factors for the development of ERF in postoperative patients with esophageal cancer and are useful in assessing the risk of developing ERF. After the occurrence of postoperative ERF, timely intervention by insertion of tracheal stents to seal the fistula may prolong the survival time of the patients.

[The influence of IGF-â on the expression of Myf5 and TGF-ß1 in medial rectus muscle of cat model with strabismus].

Objective: To observe the influence of exogenous insulin-like growth factor-Ⅰ(IGF-Ⅰ) on the expression of myocyte differentiation factor 5 (Myf5) and transforming growth factor ß1(TGF-ß1) in medial rectus muscle of cat model with strabismus. Methods: Experiment research. Twenty-seven kittens which were in sensitive period of visual development (4-6 weeks old), were randomly divided into experimental, control and blank control groups by random numbers table method. Each experimental group was further divided into 3 sub-groups (4 weeks, 8 weeks and 12 weeks) based on drug intervention time, hence 3 kittens in each sub-group. The control group and the blank control group were also divided into 3 sub-groups respectively. Exotropia treatment models were set up through surgical methods and injection of IGF-Ⅰ(0.05 ml,0.1 g/L). The internal rectus muscles of 4 weeks sub-group, 8 weeks sub-group and 12 weeks sub-group were taken respectively after the treatment model had been set up. The internal rectus muscles of the control group and the blank control group were also taken according to corresponding time. The expressions of Myf5 and TGF-ß1 were tested with immunohistochemistry staining method, and optical density analysis method were employed to measure the average optical density value. The expression of Myf5 and TGF-ß1 was analyzed by Kruskal-Wallis and Bonferroni test. The correlation between the expression of Myf5 and TGF-ß1 and the time of drug intervention was analyzed by simple linear regression. Results: (1) In the experimental group, the expression of the Myf5 of the 4 weeks sub-group, 8 weeks sub-group and 12 weeks sub-group were 33.34±17.16, 39.24±15.25 and 47.70±19.39, which were higher than the control group (21.30±7.44, 19.43±4.75, 4.82±2.66) and the blank control group (18.95±6.59, 18.00±7.29, 5.86±2.61) at the same time point, and the differences were statistically significant in 8 weeks sub-group and 12 weeks sub-group (χ(2)=21.864, 31.814, both P<0.01). The expression of Myf5 in the experimental group increased with the extension of IGF-Ⅰ intervention time (R(2)=0.99, P<0.05). But there were negative correlation between expression of Myf5 and drug intervention times in the control group and the blank control group (R(2)=0.81, 0.80, both P<0.05). (2) In the experimental group, the expression of the TGF-ß1 of the 4 weeks sub-group, 8 weeks sub-group and 12 sub-weeks group were 0.80±0.12, 0.53±0.09, 0.42±0.08, which were higher than the control group (1.91±0.23, 2.30±1.03, 1.82±0.72) and the blank control group (2.01±0.31, 2.62±1.11, 1.83±0.67) at the same time point, and the differences were statistically significant (χ(2)=30.801, 40.278, 35.177, all P<0.01). The expression of TGF-ß1 in the experimental group decreased with the extension of IGF-Ⅰintervention time (R(2)=0.83, P<0.05). The average optical density value regression equation of TGF-ß1 in the sterile water control group and the blank control group was 0.04 and 0.06, respectively, and the fitting degree was very poor. Therefore, there was no correlation trend with time. Conclusions: Exogenic IGF-Ⅰ could enhance the expression of Myf5 in medial rectus muscle of cat model with strabismus. Exogenic IGF-Ⅰ could inhibit the expression of TGF-ß1 in medial rectus muscle of cat model with strabismus. Repeated injection of exogenous IGF-Ⅰ may continuously enhance the expression of Myf5 and inhibit the expression of TGF-ß1. (Chin J Ophthalmol, 2018, 54: 375-382).

A linewidth-narrowed and frequency-stabilized dye laser for application in laser cooling of molecules.

We demonstrate a robust and versatile solution for locking the continuous-wave dye laser for applications in laser cooling of molecules which need linewidth-narrowed and frequency-stabilized lasers. The dye laser is first stabilized with respect to a reference cavity by Pound-Drever-Hall (PDH) technique which results in a single frequency with the linewidth 200 kHz and short-term stabilization, by stabilizing the length of the reference cavity to a stabilized helium-neon laser we simultaneously transfer the ± 2 MHz absolute frequency stability of the helium-neon laser to the dye laser with long-term stabilization. This allows the dye laser to be frequency chirped with the maximum 60 GHz scan range while its frequency remains locked. It also offers the advantages of locking at arbitrary dye laser frequencies, having a larger locking capture range and frequency scanning range to be implemented via software. This laser has been developed for the purpose of laser cooling a molecular magnesium fluoride beam.

Molecular characterization of pyraclostrobin resistance and structural diversity of the cytochrome b gene in Botrytis cinerea from apple.

Botrytis cinerea isolates obtained from apple orchards were screened for resistance to the quinone outside inhibitor (QoI) pyraclostrobin. Of the 220 isolates tested, 43 (19.5%) were resistant to pyraclostrobin. Analysis of partial sequences of the cytochrome b gene (cyt b) in five pyraclostrobin-resistant (PR) and five pyraclostrobin-sensitive (PS) isolates showed that PR isolates harbored the point mutation leading to the substitution of glycine by alanine at codon position 143 in cyt b (G143A). Two pairs of allele-specific primers were designed based on this point mutation, and allele-specific polymerase chain reaction analysis with these primers showed that all 73 PR isolates (including 30 collected from decayed apple fruit) harbored the G143A mutation but PS isolates did not. Six pairs of primers were designed to analyze the presence of various introns in cyt b. There were six types (I to VI) of cyt b present in 247 isolates of B. cinerea collected from various apple-production areas in Washington State. Of the 247 isolates, 23 had type I cyt b containing all four introns (Bcbi-67/68, Bcbi-131/132, Bcbi-143/144, and Bcbi-164), 176 had type II cyt b containing three introns (Bcbi-67/68, Bcbi-131/132, and Bcbi-164), six had type III cyt b containing two introns (Bcbi-67/68 and Bcbi-131/132), one had type IV cyt b containing two introns (Bcbi-131/132 and Bcbi-164), one had type V cyt b containing only the Bcbi-131/132 intron, and 40 had type VI cyt b containing no introns. This is the first report of types III to VI cyt b present in B. cinerea. All 73 PR isolates did not carry the Bcbi-143/144 intron in cyt b. Of the 247 isolates tested, >90% did not carry the Bcbi-143/144 intron in cyt b, suggesting that B. cinerea populations from apple pose a high inherent risk for the development of resistance to QoIs because the presence of this intron in cyt b prevents the occurrence of G143A-mediated resistance. Analysis of genetic background based on three microsatellite primers showed that PR isolates originated from different lineages, and there was no correlation between cyt b types (I, II, and III) and the genetic background of the isolates; however, isolates carrying type VI cyt b might originate from the same lineage.

Molecular characterization of boscalid resistance in field isolates of Botrytis cinerea from apple.

Botrytis cinerea isolates obtained from apple orchards were screened for resistance to boscalid. Boscalid-resistant (BosR) isolates were classified into four phenotypes based on the levels of the concentration that inhibited fungal growth by 50% relative to control. Of the 220 isolates tested, 42 were resistant to boscalid, with resistant phenotypes ranging from low to very high resistance. There was cross resistance between boscalid and carboxin. Analysis of partial sequences of the iron-sulfur subunit of succinate dehydrogenase gene in B. cinerea (BcSdhB) from 13 BosR and 9 boscalid-sensitive (BosS) isolates showed that point mutations in BcSdhB leading to amino acid substitutions at the codon position 272 from histidine to either tyrosine (H272Y) or arginine (H272R) were correlated with boscalid resistance. Allele-specific polymerase chain reaction (PCR) analysis of 66 BosR isolates (including 24 additional isolates obtained from decayed apple fruit) showed that 19 carried the point mutation H272Y and 46 had the point mutation H272R, but 1 BosR isolate gave no amplification product. Analysis of the BcSdhB sequence of this isolate revealed a different point mutation at codon 225, resulting in a substitution of proline (P) by phenylalanine (F) (P225F). The results indicated that H272R/Y in BcSdhB were the dominant genotypes of mutants in field BosR isolates from apple. A multiplex allele-specific PCR assay was developed to detect point mutations H272R/Y in a single PCR amplification. Levels of boscalid resistance ranged from low to very high within isolates carrying either the H272R or H272Y mutation, indicating that, among BosR isolates, different BosR phenotypes (levels of resistance) were not associated with particular types of point mutations (H272R versus H272Y) in BcSdhB. Analysis of genetic relationships between 39 BosR and 56 BosS isolates based on three microsatellite markers showed that 39 BosR isolates and 30 BosS isolates were clustered into two groups, and the third group consisted of only BosS isolates, suggesting that the development of resistance to boscalid in B. cinerea likely is not totally random, and resistant populations may come from specific genetic groups.