Objective: To investigate the effect of neurite outgrowth inhibitor extracellular peptide residues 1-40 (NEP1-40) combined with poly (lactic-co-glycolic acid) (PLGA) and gelatin electrospun fiber membrane on facial nerve repair in rats. Methods: According to the principle of random grouping, 108 male SD rats were divided into four groups (n = 27 in each group, approved by the ethics committee), namely, the sham group, control group, PLGA group, and NEP1-40 + PLGA group. A facial nerve fracture model was established for all of the groups except for the sham group. The control group received no further treatment, the PLGA group and the NEP1-40+PLGA group were supported by PLGA membrane, and the NEP1-40+PLGA group received one immediate local injection of NEP1-40 (5 μg/μL) at a dose of 10 μL. Facial nerve function analysis, electrophysiological examination, transmission electron microscope observation, HE staining, and immunohistochemical staining of myelin marker S100β and axonal marker β3-tubulin were used to evaluate the recovery of injured facial nerves of rats at 2, 4 and 8 weeks. Results : At 8 weeks, the facial nerve function score of the NEP1-40+PLGA group was better than that of the control group and PLGA group (P < 0.001), and facial nerve function was significantly restored. Electrophysiological examination of nerve action potentials at the injured facial nerve showed that the amplitude in the NEP1-40+PLGA group was higher than that of the control group and PLGA group (P < 0.001), but there was no significant difference in latency and conduction velocity results between the groups (P > 0.05). At 2, 4, and 8 weeks, transmission electron microscopy showed that the number of myelinated nerve fibers and myelin sheath thickness in the cross-section of the injured facial nerve in the NEP1-40+PLGA group were greater than those in the other groups (P < 0.05). At 8 weeks, HE staining showed that the facial nerves in the control group had partially recovered, but the overall cell distribution was uneven and the boundary with surrounding tissues was slightly blurred. In contrast, the NEP1-40+PLGA group had a relatively uniform cell distribution and a clearer boundary with surrounding tissues. At 2, 4, and 8 weeks, the immunohistochemical results showed that in the cross-section of the injuried facial nerve, NEP1-40 increased the expression of neural markers S100 β and β3-tubulin, especially β3-tubulin, which was close to normal levels (P > 0.05) Conclusion NEP1-40 is beneficial for the generation of new myelin sheaths and axons at the site of injury, and it can promote the repair and regeneration of injured facial nerves to a certain extent, thus accelerating the recovery of injured nerve function.

African swine fever(ASF),caused by the African swine fever virus(ASFV),is a highly contagious infectious disease of pigs.This disease has been spread rapidly in China since 2018,po-sing a huge threat to China's pig farming industry.To rapid detect the ASFV,a loop-mediated iso-thermal amplification(LAMP)combined with the disposable nucleic acid visualization test strip was established for visual detection of the nucleic acid of ASFV B646L gene.The method was easy to operate without special instruments and equipment,while it effectively avoided the disadvantage of false positives caused by aerosol contamination.The method was able to detect 1.16 copies/μL of the recombinant plasmid in 50 min at 65 ℃.In addition,the method was specific with no cross-re-action with classical swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,transmissible gastroenteritis virus.The results in this study provides a rapid,con-venient,sensitive and reliable method for early diagnosis and screening for ASFV suspected infec-tion cases.

To establish a simple,convenient,sensitive,and specific method for rapid detection of Zi-ka virus(ZIKV),the whole genome sequences of ZIKV isolated from different times and regions were analyzed.The specific primers and probes were designed based on the screened target se-quences located in the conserved region of the ZIKV NS5 gene.By combining RT-LAMP isother-mal amplification technology and immunochromatography technology,a reverse transcription loop mediated isothermal amplification nucleic acid and flow visualization strip(RT-LAMP-VF)detec-tion method for ZIKV was established.The results showed that the method had good specificity and sensitivity.When the ratio of inner,outer,and ring primers(FIP∶LF∶F3)was 4∶2∶1,the detection method can specifically detect 102 copies/pL RNA transcripts or 2.15 pfu ZIKV at 61 ℃for 45 minutes,with no cross reaction with other flaviviruses such as Japanese encephalitis virus and classical swine fever virus.Other RNAs in blood tissue samples did not affect the sensitivity and specificity of RT-LAMP-VF,indicating that the method can be applied to clinical practice.The ZIKV RT-LAMP-VF detection method established in this study is easy to perform and does not require special instruments and equipment.It is particularly suitable for the rapid detection of ZIKV in grassroots units,providing technical support and material support for the establishment of on-site rapid detection and early warning and prediction systems for ZIKV disease.

The PCR-amplified severe fever with thrombocytopenia syndrome virus(SFTSV)Gn-DⅢ-Ⅲ gene was inserted into the pET-30a(+)prokaryotic expression vector to generate the re-combinant plasmid pET-SFTSV-Gn-D Ⅲ-Ⅲ.The plasmid was transformed into E.coli BL21(DE3)for Gn-DⅢ-m protein expression and the expression conditions were optimized.The Gn-DⅢ-Ⅲ protein purified with Ni-NTA column affinity chromatography was applied as the captured antigen to establish an indirect ELISA method for the detection of SFTSV antibody.The results demonstrated that the recombinant plasmid pET-SFTSV-Gn-D Ⅲ-Ⅲ was successfully constructed as identified by PCR and sequencing.The recombinant protein SFTSV Gn-D m-Ⅲ was soluble ex-pression in E.coli under the optimal induction conditions of 0.4 mmol/L IPTG at 25 ℃ for 4 h,and the protein purity was 91.77％after purification by Ni-NTA column.The optimal reaction con-ditions for the indirect ELISA of SFTSV antibody were as follows:coating antigen concentration(5 μg/mL),primary antibody(incubation at 37 ℃ for 1.5 h),and secondary antibody(diluted 1:10 000 and incubated at 37 ℃ for 1 h).The established method had no cross-reactivity with Rift Valley fever virus(RVFV),Ebola virus(EBOV),and tick-borne encephalitis virus(TBEV)posi-tive sera.The method had a high sensitivity,with P/N＞2.1 for SFTSV-positive sera diluted to 81920.Coefficients of variation for intra-and inter-batch reactions were less than 10％.Detection of four SFTSV-infected human clinical serum samples showed the serum samples from patients in re-mission were tested as positive(P/N＞2.1),while serum samples from patients with multiple or-gan failure were detected as negative(P/N＜2.1).The results indicated that the SFTSV Gn-D Ⅲ-Ⅲ protein was successfully expressed and purified,and it was used as the coating protein to estab-lish an indirect ELISA assay for SFTSV antibody,which possesses good specificity,sensitivity and reproducibility.This method might be applied to detect human SFTSV clinical serum samples.

In order to explore the effects of Coptis detoxification powder on immune suppression and inflammatory response caused by heat stress in animals,Bama pigs were used as experimental animals to establish an animal model of heat stress,and the effects of Coptis detoxification powder on transcriptional activation of TLRs mRNA in PBMC induced by heat stress and its mediated ex-pression of inflammatory factors were studied in vivo.The animal model of heat stress was estab-lished by artificial climate greenhouse,Chinese medicine"Coptis detoxification powder"was pre-pared,and its inhibitory effect on heat stress and inflammation was studied by preventive and therapeutic administration.Real-time fluorescence quantitative PCR was used to determine the transcriptional changes of HSP70,TLRs and IL-17 mRNA,and ELISA was used to determine the dynamic changes of serum IL-2,IL-6 and TNF-α.Western blot and indirect immunofluorescence(IFA)were used to determine the nuclear translocation and protein expression of p65 phosphoryl-ated protein in NF-κβ inflammatory factor pathway.The transcription of PBMC TLRs mRNA and its mediated inflammatory factor expression and function in Bama pigs under heat stress were sys-tematically analyzed.The results showed that the experimental animal model of Bama pigs under heat stress was successfully established,and heat stress significantly up-regulated the mRNA tran-scription levels of HSP70,TLR2,TLR4 and inflammatory factor IL-17,resulting in significantly enhanced expressions of serum inflammatory factors IL-2,IL-6 and TNF-α.The NF-κβ pathway significantly promoted the level of p65 nuclear phosphorylation and the expression of cytoplasmic protein.The Chinese medicine"Coptis detoxification powder"could significantly reduce the upreg-ulation effect of heat stress conditions on the transcription levels of HSP70 and TLRs mRNA,thus inhibiting the mRNA transcription of inflammatory factor IL-17 and the expression of IL-2,IL-6 and TNF-α in serum,and significantly antagonizing the transcriptional activity of NF-κβ caused by heat stress.It showed good anti-heat stress effect.The anti-inflammatory effects of Chinese veteri-nary medicine prescription"Coptis detoxification powder",its pathway of action and its effectors were investigated,the expression and secretion levels of the regulatory factors related to the path-way associated with TLRs mRNA inhibition were analyzed,and the functional effects and mecha-nism of its clinical anti-heat stress and anti-inflammatory effects were clarified,which laid a data foundation for the promotion and application of this prescription and related mechanism research.

Objective To investigate the predictive value of dynamic monitoring of transcutaneous oxygen partial pressure [p_tc(O₂)], transcutaneous carbon dioxide partial pressure [p_tc(CO₂)], serum amyloid A (SAA), and interleukin-6 (IL-6) levels in predicting bronchopulmonary dysplasia (BPD) in neonates with neonatal respiratory distress syndrome (NRDS). Methods A total of 240 NRDS patients were selected as the research subjects and divided into BPD group (108 cases) and non-BPD group (132 cases) based on whether BPD occurred. The levels of p_tc(O₂), p_tc(CO₂), SAA, and IL-6 were compared between the two groups on the 1st, 7th, and 14th day after birth. Spearman correlation analysis was used to explore the correlation between each indicator and the degree of illness. The relative risk (RR) and 95% confidence interval (CI) were used to evaluate the impact of each indicator on BPD in NRDS patients. Receiver operating characteristic (ROC) curves and decision curve analysis were used to evaluate the predictive efficacy and clinical net benefit of each indicator for BPD. Results On the 1st day after birth, there were no statistically significant differences in p_tc(O₂), p_tc(CO₂), serum SAA, and IL-6 levels between the two groups (P>0.05). On the 7th and 14th day after birth, the p_tc(O₂) in the BPD group was lower than that in the non-BPD group, while the p_tc(CO₂) and serum SAA, IL-6 levels were higher than those in the non-BPD group (P < 0.05). Correlation analysis results showed that the degree of BPD in children was negatively correlated with p_tc(O₂) on the 7th day after birth (r=-0.724, P < 0.05), and positively correlated with p_tc(CO₂), SAA, and IL-6 on the 7th day after birth (r=0.635, 0.830, 0.715, P < 0.05). The degree of pulmonary dysfunction in children was negatively correlated with p_tc(O₂) on the 7th day after birth (r=-0.719, P < 0.05), and positively correlated with p_tc(CO₂), SAA, and IL-6 on the 7th day after birth (r=0.673, 0.756, 0.696, P < 0.05). On the 7th day after birth, the p_tc(O₂) of BPD patients was negatively correlated with SAA and IL-6 (r=-0.605, -0.623, P < 0.05), while p_tc(CO₂) was positively correlated with SAA and IL-6 (r=0.618, 0.650, P < 0.05). ROC curve showed that the combined prediction of p_tc(O₂), p_tc(CO₂), SAA, and IL-6 on the 7th day after birth for NRDS complicated with BPD was better than the individual prediction of the four indicators, with an area under the curve of 0.938 (95%CI, 0.899 to 0.965). The risk of BPD in high-expression of p_tc(CO₂), SAA, and IL-6 was 2.256, 1.668, and 1.667 times higher than that in low-expression, respectively. The risk of BPD in high-expression of p_tc(O₂) was 0.568 times higher than that in low-expression. The decision curve analysis showed that when the threshold value was within the range of 0.2 to 0.9, the combined model of p_tc(O₂), p_tc(CO₂), SAA, and IL-6 was superior to the individual prediction in predicting the net benefit rate of BPD in NRDS patients on the 7th day after birth. Conclusion The severity of NRDS complicated with BPD is closely related to p_tc(O₂), p_tc(CO₂), SAA, and IL-6, and the combined detection of the four indicators can help improve the predictive efficacy and net benefit rate of NRDS complicated with BPD.

BACKGROUND/OBJECTIVES: This study examined the relationship between famine exposure in early life and the risk of type 2 diabetes in adulthood during the 1959–1961 Chinese Famine. SUBJECTS/METHODS: A total of 3,418 individuals aged 35–74 years free of diabetes from two studies in 2006 and 2009 were followed up prospectively in 2009 and 2012, respectively. Famine exposure was classified as unexposed (individuals born in 1962–1978), fetal exposed (individuals born in 1959–1961), child exposed (individuals born in 1949–1958), and adolescent/adult exposed (born in 1931–1948). A logistic regression model was used to assess the relationship between famine exposure and diabetes after adjustment for potential covariates. RESULTS: During a three-year follow-up, the age-adjusted incidence rates of type 2 diabetes were 5.7%, 14.5%, 12.7%, and 17.8% in unexposed, fetal-exposed, child-exposed, and adolescent/adult-exposed groups, respectively (P < 0.01). Relative to the unexposed group, the relative risks (95% confidence interval) for diabetes were 2.15 (1.29–3.60), 1.53 (0.93– 2.51), and 1.65 (0.75–3.63) in the fetal-exposed, child-exposed, and adolescent/adult-exposed groups, after controlling for potential covariates. The interactions between famine exposure and obesity, education level, and family history of diabetes were not observed, except for the urbanization type. Individuals living in rural areas with fetal and childhood famine exposure were at a higher risk of type 2 diabetes, with relative risks of 8.79 (1.82–42.54) and 2.33 (1.17–4.65), respectively. CONCLUSIONS These findings indicate that famine exposure in early life is an independent predictor of type 2 diabetes, particularly in women. Early identification and intervention may help prevent diabetes in later life.

Objective:To investigate the effects of mindfulness therapy on blood glucose control and quality of life in older adult patients with diabetes mellitus and cognitive impairment.Methods:A total of 313 patients with type 2 diabetes mellitus and cognitive impairment aged over 75 years who received treatment in Xinxiang First People's Hospital from August 2018 to July 2020 were included in this study. They were randomly allocated to the intervention group ( n = 157) and control group ( n = 156). All patients underwent treatments for diabetes mellitus and underlying diseases. The intervention group was subject to 8 weeks of mindfulness therapy. The percentage of patients with qualified glycosylated hemoglobin in human whole blood and 36-Item Short Form Health Survey questionnaire score were compared between the two groups. The Pearson correlation was performed. Results:After 8 weeks of treatment, the percentage of patients with qualified glycosylated hemoglobin in human whole blood in the intervention group was 43.95%, which was significantly higher than the level (25.48%) at the recruitment and the level (30.77%) in the control group ( χ2 = 12.17, P = 0.002; χ2 = 5.80, P = 0.016). The quality of life score was significantly higher in the intervention group than in the control group [(72.16 ± 14.08) points vs. (64.37 ± 15.49) points, t = 4.65, P = 0.001]. Conclusion:Mindfulness therapy can regulate blood glucose level and improve quality of life in older adult patients with diabetes mellitus and cognitive impairment.

Objective:To investigate the clinicopathological features and clinical management of primary extragonadal germ cell tumor of the prostate.Methods:Two cases of primary extragonadal germ cell tumor in the prostate were collected at Fudan University Shanghai Cancer Center, in January and September 2016, respectively. Their pathological features, clinical treatments and follow-up were retrospectively analyzed.Results:The two patients were 41 and 32 years old, respectively, and both presented with obstructive symptoms of the lower urinary tract. Histologically, both cases showed small round blue cells and an invasive growth pattern. The immunohistochemistry (IHC) stains of SALL4, OCT3/4, CD117 and PLAP were all positive, while those of PSA, AR and syn were negative. Moreover, case 1 demonstrated perinuclear dot-like staining for CKpan, which might be a diagnostic pitfall. There was no evidence of disease in other areas via physical examination or radiographic studies. Based on these IHC findings and the morphology, the two cases were diagnosed as primary seminoma of the prostate, which were consequently treated with six cycles of bleomycin, etoposide and cisplatin-based chemotherapy. A complete response was achieved in case 1. Case 2 was followed up and showed tumor recurrence, and progression with elevated tumor marker AFP. The subsequent radical removed specimens of case 2 were finally diagnosed as mixed germ cell tumor of the prostate.Conclusion:As a rare neoplastic entity, primary germ cell tumor of the prostate can show small blue round cell morphology. Pathologically, the morphology of small round blue cells combined with a perinuclear dot-like pattern of CKpan IHC staining may be a diagnostic pitfall. The clinical treatment strategy should be evaluated with consideration of the pathological diagnosis and comprehensive evaluation of the tumor markers.

Objective:To evaluate the effect of definitive radiotherapy with different doses on overall survival (OS) and identify the prognostic factors of patients with non-metastatic esophageal squamous cell carcinoma (ESCC).Methods:Clinical data of 2 344 ESCC patients treated with definitive radiotherapy (RT) alone or chemoradiotherapy from 2002 to 2016 in 10 hospitals were collected and analyzed retrospectively. After the propensity score matching (PSM)(1 to 2 ratio), all patients were divided into the low-dose group (equivalent dose in 2 Gy fractions, EQD 2Gy<60 Gy; n=303) and high-dose group (EQD 2Gy≥60 Gy; n=606) based on the dose of radiation. Survival analysis was conducted by Kaplan- Meier method. Multivariate prognostic analysis was performed by Cox′s regression model. Results:The median follow-up time was 59.6 months. After the PSM, the 1-, 3- and 5-year overall survival (OS) rate was 66.5%, 34.7%, 27.2% in the low-dose group, 72.9%, 41.7% and 34.7% in the high-dose group, respectively ( P=0.018). The 1-, 3-and 5-year progression-free survival rate was 52.2%, 27.2%, 23.1% in the low-dose group, 58.3%, 38.1% and 33.9% in the high-dose group, respectively ( P=0.001). The outcomes of univariate analysis indicated that cervical/upper esophagus location, early (stage Ⅱ) AJCC clinical stage, node negative status, tumor length ≤5 cm, receiving intensity-modulated radiation therapy (IMRT), receiving concurrent chemotherapy and EQD 2Gy≥60 Gy were closely associated with better OS (all P<0.05). Multivariable analysis demonstrated that tumor location, regional lymph node metastasis, concurrent chemotherapy and EQD 2Gy were the independent prognostic factors for OS (all P<0.05). Conclusion:Three-dimensional conformal or IMRT with EQD 2Gy≥60 Gy yields favorable survival outcomes for patients with locally advanced ESCC.