Objective To establish a genetic monitoring method for laboratory quails.Methods Quail microsatellite loci were searched in the literature,and microsatellite DNA loci suitable for quails were screened by an interspecific transfer method in closely related species,namely chickens and ducks.Quail liver DNA was extracted as a template,and the corresponding loci were screened by PCR amplification and agarose gel electrophoresis.On the basis of amplification of the selected microsatellite loci,the number of alleles,polymorphisms,and microsatellite loci combinations for quail genetic quality detection were selected and detection method were developed.Results We preliminary determined 23 microsatellite loci for genetic monitoring of closed-colony laboratory quails.Conclusions A genetic monitoring method for laboratory quails was preliminary developed.

Objective To investigate the expression level of serum ubiquitin-conjugating enzyme 2C(UBE2C)and tripartite motif-containing protein 27(TRIM27)in patients with endometrial cancer(EC)and their correlation with pathological parameters.Methods A total of 96 EC patients from March 2020 to March 2023 were selected as EC group;65 patients with endometrial atypical hyperplasia(EAH)as EAH group,and 80 healthy subjects as the control group.Enzyme linked immunosorbent assay was applied to analyze the expression level of serum UBE2C and TRIM27.The relationship between serum UBE2C,TRIM27,and pathological data was analyzed;receiver operating characteristic was applied to evaluate the predictive value of serum UBE2C and TRIM27 level for EC.Results The level of serum UBE2C and TRIM27 of EC patients was obviously higher than that in healthy subjects(P<0.05),and was correlated with tumor diameter,tumor differentiation,lymph node metastasis,FIGO stage,muscle layer invasion depth,cervical involvement,estrogen receptor expression,and progestogen receptor expression(P<0.05).There was positive correlation between serum UBE2C and TRIM27(r=0.475,P<0.001);and the level of serum UBE2C and TRIM27 was positively correlated with tumor diameter,lymph node metastasis,FIGO stage,and depth of musclular invasion,but negatively with tumor differentiation,estrogen receptor expression,and progestogen receptor expression(P<0.05).The combination of UBE2C and TRIM27 had obviously higher AUC in evaluating EC than single detection(ZUBE2C-combination=3.406,P<0.001,ZTRIM27-combination=3.285,P=0.001).Conclusion The expression level of UBE2C and TRIM27 in serum of EC patients is up-regulated,which is closely related to pathological parameters.The level of serum UBE2C and TRIM27 can provide reference for early diagnosis of EC.

Objective A HPLC-quantitative analysis of multi-components by single marker(QAMS)was established to determine 5 ingredients including uracil,uridine,hypoxanthine,inosine and guanosine in Periplaneta americana.Methods Separation took place on a Agilent ZORBAX SB-Aq column(250 mm×4.6 mm,5 μm)by gradient elution of methanol-0.01 mol·L-1 potassium dihydrogen phosphate at 20℃with a flow rate of 1.0 mL·min-1.The detection wavelength was 260 nm and the injection amount was 10 μL.The relative correction factors(fa/b)was calculated for the other four components with uridine as an internal standard.The content of 5 ingredients in 10 batches of Periplaneta americana was determined by QAMS.Results were compared with those of external standard method(ESM).Results Five nucleosides showed good linear relationships in their own ranges(r＞0.999 5),and the average recoveries ranged from 97.0%to 100.8%.The relative correction factors of uracil,hypoxanthine,inosine and guanosine were 0.908 0,1.005 3,1.969 5 and 1.303 4,respectively.Conclusion The established method is accurate and stable.It can provide theoretical reference for the quality control of Periplaneta americana.

OBJECTIVE To establish HPLC characteristic chro matogram of Jianpi yifei biyan prescription standard decoction ， to select the quality control index components and determine their contents. METHODS HPLC method combined with Similarity Evaluation System of TCM Chromatographic Fingerprint （2004 edition）were used to establish the characteristic chromatogram of 10 batches of Jianpi yifei biyan prescription standard decoction ；the similarity evaluation and common peaks identification were also carried out. Using common peak area of characteristic chromatogram as variables ，SPSS 26.0 software and SIMCA 14.1 software were used to perfor m cluster analysis （CA），principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA）；differential components with variable important i n pro jection（VIP）value greater than 1.5 were screened；the contents of cimifugin and differential components were determined by the same method. RESULTS A total of 24 common characteristic peaks were identified ， and the similarities of 10 batches of samples were higher than 0.960；eight characteristic peaks were identified by comparison with reference substance. CA and PCA results revealed that the samples were classified into 3 categories.OPLS-DA analysis showed that 3 components with VIP value greater than 1.5， which were prim-O-glucosylcimifugin （peak 2），calycosin 7-O-β-D-glucopyranoside （peak 4） and 5-O-methylvisammioside （peak 6） in descending order. The linear ranges of prim- O- glucosylcimifugin，calycosin 7-O-β-D-glucopyranoside，cimifugin and 5-O-methylvisammioside were 0.010 7-0.213 0，0.007 8- 0.156 0，0.008 0-0.160 0，0.009 8-0.195 0 μg（r＞0.999），respectively. RSD values of precision ，repeatability and stability tests （24 h） were all less than 2％. Average recoveries were 105.98％（RSD＝1.75％，n＝6），98.06％（RSD＝3.87％，n＝6），96.38％（RSD＝ 4.03％ ，n＝6） and 104.17％（RSD＝1.27％ ，n＝6）. The contents of the above 4 components in 10 batches of samples were 12.12-18.87，3.86-6.40，3.10-4.27 and 11.17-15.79 μ g/mL，respectively. CONCLUSIONS The established HPLC characteristic chromatographic method is stable and feasible ，it can be used for the quality control of Jianpi yifei biyan prescription standard decoction. Prim- O-glucosylcimifugin，calycosin 7-O-β-D-glucopyranoside，cimifugin and 5-O-methylvisammioside can be used as the index components for quality control of the standard decoction.

Objective:To study the effect of Sanjie Xiaoliu Recipe on transplanted tumors in breast cancer mice through in vivo experiments, in order to explore the efficacy and mechanism of Sanjie Xiaoliu Recipe on breast cancer patients. Methods:45 BaL B/c female mice were selected to establish the transplanted tumor model of breast cancer. All the transplanted tumor models of breast cancer mice were randomly divided into five groups: the control group (intragastric administration of normal saline), the low, medium and high dose group of Sanjie Xiaoliu Recipe (intragastric administration of different doses of Sanjie Xiaoliu Decoction), and the paclitaxel group (intraperitoneal injection of paclitaxel). After 24 days of continuous administration, the diet and activities of mice were observed; the body weight, weight and volume changes of transplanted tumor mice were recorded, and the tumor inhibition rate was calculated. Western blot was used to detect the expression of epithelial mesenchymal transformation related proteins (E-cadherin, N-cadherin, Vimentin) in the transplanted tumor tissues of mice in each group.Results:(1) The food intake and activity status of mice treated with Sanjie Xiaoliu Recipe were less affected by the transplanted tumor of breast cancer. (2) The volume and weight of transplanted tumor in the treat groups were smaller than those in the control group (all P<0.01), and the volume of transplanted tumor in the middle dose group of Sanjie Xiaoliu Recipe was smaller than that in the low dose group ( P<0.05). The tumor inhibition rates among the treatment groups were: Sanjie Xiaoliu Recipe medium dose group 52.4%, paclitaxel group 40.3%, Sanjie Xiaoliu Recipe low dose group 39.5%, Sanjie Xiaoliu Recipe high dose group 34.1%. (3) The results of Western blot showed that the expression level of E-cadherin in the transplanted tumor tissue of the treat groups was higher than that in the control group, and the expression levels of N-cadherin and Vimentin were lower than that in the control group, with statistically significant difference (all P<0.05). Conclusions:The Sanjie Xiaoliu Recipe can improve the weakness and reduced consumption of breast cancer mice, which can inhibit the tumor mass growth in mice to a certain extent. Its mechanism may be that Sanjie Xiaoliu Recipe can inhibit the epithelial mesenchymal transformation of breast cancer and the invasion and metastasis of breast cancer.

Closed-loop hospital management can effectivly cope with the COVID-19 pandemic. In order to ensure the continuity of treatments for hemodialysis patients under closed-loop management and minimize possible medical and infection risks, Huashan Hospital affiliated to Fudan University and 9 hospitals in Shanghai established a hemodialysis alliance in January 2021.The alliance optimized hemodialysis resources within the region through overall planning by preparing sites, materials and personnel shifts in advance, and establishing management systems and work processes to ensure that patients could be quickly and orderly diverted to other blood dialysis centers for uninterrupted high-quality hemodialysis services, in case that some hemodialysis centers in the alliance under closed-loop management.From November 2021 to April 2022, 317 of 1 459 hemodialysis patients in the alliance were diverted to other centers for treatment, accumulating 1 215 times/cases of treatments without obvious adverse reactions. The practice could provide a reference for medical institutions to quickly establish mutual support mode under major public health events.

Objective:To investigate the multilocus sequence typing feature of the virulence-associated genes of Staphylococcus aureus(S. aureus) separated from the clinical specimens of a multi-center cohort children in Guangzhou area. Methods:A total number of 412 Staphylococcus aureus strains isolated from 2 059 non-repeated fecal specimens of children by three groups′ researchers in Guangzhou Women and Children′s Medical Center from August 2018 to November 2018. While collecting specimens, patient clinical information is also properly collected and preserved. After extracting the DNA of the strain, the virulence-associated genes were detected by polymerase chain reaction (PCR), including the staphylococcal enterotoxin (SE) genes ( sea, seb, sec, sed, see) and the Panton-Valentine leucocidin-encoding gene ( pvl).The multi-locus sequence typing (MLST) method was performed to reveal the MLST feature of these genes and the statistical difference were examined by the the χ 2 test. Results:Among the 412 isolates of S. aureus, 256 strains (256/412, 62.1%) contains at least one SE gene. Among the enterotoxin gens, the sec (125/412, 30.3%), seb(98/412, 23.8%)and sea (66/412, 16.0%)genes were the three most prevalent members of SEs. The frequency of pvl gene in Staphylococcus aureus was 18.7%(77/412).Among them, the frequency of Staphylococcus aureus sea gene isolated from patients with gastroenteritis (58/319, 18.2%) was significantly higher than that from the non-gastroenteritis group (8/93, 8.6%)(χ2=4.912, P=0.027). The frequency of Staphylococcus aureus pvl gene isolated from the patients with pneumonia (8/21, 38.1%) was greater than that from the non-pneumonia group (6/47, 12.8%)(χ2=4.252, P=0.039). In addition, the virulence-associated gene of S. aureus was closely related to the specific ST type, 82.4% (28/34) of ST6 carried sea gene, all ST338 and ST59 carried seb gene, 96% (48/50) ST45 carried sec gene, and the pvl gene carrying rate of ST338 was 5/5. Conclusions:The SEA toxin produced by ST6 Staphylococcus aureus may be closely related to the diagnosis of gastroenteritis in children. The frequency of pvl virulence gene in Staphylococcus aureus in children with community-acquired pneumonia was higher than that in the non-pneumonia group, and closely related to the CC59.

Objective:To retrospectively assess the relationship between immune disorder and acute gastrointestinal injury (AGI) in patients after severe polytrauma.Methods:Totally 205 patients with severe polytrauma admitted to Tongji Hospital from April 2018 to October 2019 were enrolled as the observation group, and 23 healthy volunteers were served as the control group. According to the diagnostic criteria of AGI, all patients were divided into the AGI group (with AGI) or N-AGI group (without AGI), AGI patients were divided into the S-AGI group or L-AGI group according to the severity. The levels of cytokines and lymphocyte subset were evaluated at day 1, 7, and 14 after severe polytrauma. The differences between groups were statistically analyzed. The independent risk factors of AGI were analyzed by Logistic regression analyzed.Results:Totally 79.5% (163/205) of patients with severe polytrauma were accompanied by AGI. There were significant differences in the ratio of Tc, Th at day 1 after trauma, the levels of IL-6, TNF-α, IL-8, IL-10, the ratio of Ts, Th/Ts, Treg at day 7 after trauma, and the levels of IL-8, IL-10,the ratio of Ts, Th/Ts, Treg at day 14 after trauma between the AGI group and N-AGI group ( P<0.05). There were significant differences in the ratio of Tc, Th, the levels of IL-6, TNF-α at day 1 after trauma and the ratio of Ts, Th/Ts, Treg, the levels of IL-8, IL-10 at day 7 and 14 after trauma between the S-AGI group and L-AGI group ( P<0.05). Logistic regression analysis showed that Ts 7 d ( OR=2.018, 95% CI: 1.105-5.364, P=0.013), Treg 14 d ( OR=3.612, 95% CI: 1.375-8.476, P=0.006), IL-6 7 d ( OR=1.824, 95% CI: 1.011-5.835, P=0.024), IL-10 14 d ( OR=2.847, 95% CI: 1.241-6.216, P=0.014), TNF-α 7 d ( OR=1.754, 95% CI: 1.215-5.441, P=0.018) were independent risk factors in patients with AGI after severe polytrauma. Conclusions:AGI is more easily occurred in patients with the heavier immune disorders after severe polytrauma. AGI can also aggravate pre-existing immune disorders in patients after severe polytrauma.

Objective:To retrospectively assess the occurrence and risk factors in patients with acute gastrointestinal injury (AGI) after polytrauma.Methods:Totally 430 patients with polytrauma admitted to Tongji Hospital from April 2018 to October 2019 were enrolled as the observation group. According to the diagnostic criteria of AGI, all patients were divided into the AGI group (with AGI) or N-AGI group (without AGI). The patients with abdominal injury or previously suffered from gastrointestinal disease were excluded. The patient's clinical characteristics lab tests results, and the first ISS, APACHEⅡ, SOFA and GCS scores were collected. The differences between different groups were statistically analyzed. The independent risk factors of AGI were analyzed by Logistic regression.Results:65.3% of patients with polytrauma were accompanied by AGI (281/430 cases).There were significant differences between the AGI group and N-AGI group in ISS, GCS, APACHE Ⅱ and SOFA score, PCT or IL-6 level, shock index and length of stay in ICU ( P<0.05). Logistic regression analysis showed that shock, ISS≥16, APACHE Ⅱ≥16, SOFA≥5, GCS≤8 and IL-6>50 pg/mL were the early independent risk factors in patients with ACI after polytrauma. Conclusion:The incidence of AGI in patients after polytrauma is higher, which is related to ischemia, hypoxia, abnormal blood coagulation and stress in the early stage after trauma.

Objective:To observe the pronunciation of consonants among children with developmental speech sound disorder and explore the correlation between mispronounced consonants and short-term memory so as to determine the pathogenesis of the disorder.Methods:Thirty-six children with developmental speech sound disorder and aged 4 to 13 years were evaluated. Their pronunciation of consonants at the phoneme and lexical levels was tested to record the error types and error rate. Twelve of the children were then randomly chosen to form a voice disorder group. Another 10 healthy counterparts constituted a control group. The short-term memory of both groups was assessed and any correlation between pronunciation and short-term memory was analyzed.Results:The children with a developmental speech sound disorder differed significantly from the controls in terms of the numbers of errors in articulating blade-alveolar, blade-palatal and velar consonants. On the phoneme level, the highest substitution error rate occurred when pronouncing lingua-palatal consonants (42.86%), followed by supradental consonants (32%). The highest distortion and non-acquisition error rates were with blade-palatal consonants (14%) and lingua-palatal consonants (9.5%). On the vocabulary level, the highest substitution, distortion, ellipsis and non-acquisition error rates appeared when pronouncing lingua-palatal and velar consonants, velar and blade-palatal consonants, supradental consonants as well as blade-palatal consonants. Significant differences were found between the phoneme and lexical levels in the substitution of supradental and blade-palatal consonants as well as in the ellipsis of blade-alveolar consonants. They were moderately associated with pronunciation level. There was, however, no significant difference in working memory span between the two groups, and no significant correlation was observed between working memory span and pronunciation level.Conclusion:The mispronunciation of consonants by children with developmental speech disorders is higher at the lexical than at the phoneme level. They mainly substitute lingua-palatal and velar consonants and elide supradental consonants, which may be related to short-term memory span.