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.

Moloney leukemia virus 10(MOV10)gene knockout(MKO)mouse neuro 2a(N2a)cell lines was constructed by CRISPR/Cas9(clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9)gene editing technology.First,a recombinant plasmid pMD18T-U6-sgRNA expressing MOV10 gene-specific guide RNA(sgRNA)was constructed,and then pMD18T-U6-sgRNA and pMJ920-Cas9-eGFP were co-transfected into N2a.The results showed that the MKO N2a cell line had normal cell activity and cell proliferation ability.The infection test of the MKO N2a cell line was carried out using the rabies virus(RABV)and vesicular stomatitis virus(VSV)of the Rhabdoviridae family.The results showed that the replication level of the Rhabdoviridae virus in the MKO N2a cell line was significantly enhanced.The results showed that a MKO N2a cell line was successfully constructed in this study,which provided a preliminary basis for the exploration of the biological function and antiviral mechanism of MOV10 and the develop-ment of a recombinant viral vector vaccine with RABV/VSV as the vector.

Moloney leukemia virus 10(MOV10)gene knockout(MKO)mouse neuro 2a(N2a)cell lines was constructed by CRISPR/Cas9(clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9)gene editing technology.First,a recombinant plasmid pMD18T-U6-sgRNA expressing MOV10 gene-specific guide RNA(sgRNA)was constructed,and then pMD18T-U6-sgRNA and pMJ920-Cas9-eGFP were co-transfected into N2a.The results showed that the MKO N2a cell line had normal cell activity and cell proliferation ability.The infection test of the MKO N2a cell line was carried out using the rabies virus(RABV)and vesicular stomatitis virus(VSV)of the Rhabdoviridae family.The results showed that the replication level of the Rhabdoviridae virus in the MKO N2a cell line was significantly enhanced.The results showed that a MKO N2a cell line was successfully constructed in this study,which provided a preliminary basis for the exploration of the biological function and antiviral mechanism of MOV10 and the develop-ment of a recombinant viral vector vaccine with RABV/VSV as the vector.

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.

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: The therapeutic effect of poly(ADP-ribose) polymerase inhibitors (PARPi) monotherapy compared with platinum-based chemotherapy, and the impact to subsequent platinum-based chemotherapy after PARPi resistance were inconclusive. Methods: BRCA1/2 mutant ovarian cancer patients with secondary platinum-sensitive relapse were included. The patients did not receive any maintenance regimen after first- and second-line platinum therapy, and the secondary platinum-free interval (PFI) was more than 6 months. Patients in study group were treated with PARPi monotherapy until disease progression, and patients in control group were treated with platinum-based chemotherapy. Results: A total of 64 patients were retrospectively analyzed, including 31 (48.4%) in study group and 33 (51.6%) in control group. The objective response rate (77.4% vs. 84.0%; p=0.538) and median progression-free survival (8.6 vs. 11.1 months; p=0.679) were comparable. PARPi monotherapy significantly prolonged post-recurrent survival (PRS) (hazard ratio [HR]=0.35; p=0.024), and was the independent factor associated with PRS (HR=0.33; p=0.038) in multivariate analysis. The median time from treatment to first subsequent therapy or death (mTFST) of patients with platinum-based chemotherapy after PARPi progression and patients in control group with PFI ≥6 months after third-line platinum-based chemotherapy was comparable (mTFST: 7.5 vs. 7.1 months; p=0.800). Further survival analysis showed that PRS of patients with PARPi monotherapy were similar to patients with PFI ≥6 months after third-line platinum chemotherapy (HR=0.66; p=0.503), and superior to patients with PFI <6 months after third-line platinum chemotherapy (HR=0.15; p=0.009). Conclusion PARPi monotherapy was equivalent to platinum-based chemotherapy for BRCA1/2-mutated ovarian cancer patients with secondary platinum-sensitive recurrence, and could improve prognosis.

Objective To investigate the intrauterine growth characteristics of twins and birthweight discordant twins (discordant twins ).MethodsTotal of 1010 twin pregnancies (2020 fetuses) with complete delivery records from the Department of Obstetrics and Gynecology,the First and Third Affiliated Hospital of SUN Yat-sen University between January 1,2000 and July 31,2010 were studied retrospectively.One handred and ninteen cases (238 fetuses) with intrapair birthweight difference ≥25％ were determined as the discordant twins group,and the other 891 cases (1782 fetuses) with intrapair birthweight difference ＜ 25％ were identified as the concordant twins group.The singleton control group included 4042 singleton pregnancies in the same period.Results ( 1 ) Comparison of clinical data between the twins groups:the birthweight of larger-twin,smaller-twin and intrapair birthweight difference in the discordant twins group and the concordant twins group were ( 2090± 827 ) g,( 1392 ± 592 ) g,( 33.9 ±9.3 ) ％,and ( 2408 ± 543 ) g,( 2191 ± 505 ) g,( 8.9 ± 6.5 ) ％,respectively,with significant differences (P＜0.01).The incidence of discordant twins was 11.78％ (119/1010).Compared with the concordant twins group,the discordant twins group bad higher proportion of monochorionic twins,and higher prevalence of pregnancy complications such as late miscarriage,abnormal umbilical insertion,twin-twin transfusion syndrome and hypertensive disorders in pregnancy ( P ＜ 0.05 ).( 2 ) The characteristics of twin birthweight distribution:① In all the 2020 twins,80.05％ (1617/2020) fetuses had birthweight below the 50th percentile of the singleton control group,while 23.71％ (479/2020) feeuses got birthweight below the 10th percentile of the singleton control group.② After 19th gestational week,the 50th and 90th percentile of all twins' birthweight were lower than those of singletons.After 38th gestational week,the birthweight of singletons kept increasing and reached its peak at 41 th week,while the birthweight of twins reached its peak at 38th week,followed by a decline at 39 weeks,which was even lower than the 10th percentile of the singleton control group.③ The distribution of birthweight of larger- and smaller-twin in the discordant twins group:65 (54.6％,65/119) larger-twins and one (0.8％,1/119) smaller-twin had birthweight above the 50th percentile of all twins,while 5 (4.2％,5/119) larger-twins and 97 ( 81.5％,97/119 ) smaller-twins got birthweight below the 10th percentile of all twins.Conclusions ( 1 ) The patterns of birthweight curves for each gestational week are different between twins and singletons.In order to evaluate the growth of twins, birthweight reference for twins shoull be employed.( 2 ) According to the reference of twins birthweight,the most discordant twins are complicated with fetal growth restriction at least in one twin.