Objective To establish an animal model of hand,foot,and mouth disease(HFMD)in Syrian hamsters coinfected with coxsackievirus A6(CVA6)and coxsackievirus B1(CVB1).Methods 42 Syrian hamsters were divided into a CVA6 infection group,CVB1 infection group,CVA6 and CVB1 coinfection group and control group.A HFMD model was established by nasal instillation of virus solution and phosphate-buffered saline.Clinical and physiological indicators and detoxification status were monitored and recorded for 15 d,and animals were selected on day 7(D7)after infection for histopathology and viral antigen and nucleic acid testing.Results Hamsters in the single-infection and coinfection groups showed clinical symptoms similar to human HFMD.White blood cell,neutrophil,and lymphocyte result were characteristic of viral infection.Both viral nucleic acids were detected in throat swabs,feces,blood,and tissues and both viruses were isolated from fecal samples.Pathological damage and positive co-localization of CVA6 and CVB1 viral antigen proteins and nucleic acids were found in brain and other tissues.Conclusions Nasal instillation of a CVA6 and CVB1 mixture can successfully coinfect Syrian hamsters,replicate herpes infection similar to human HFMD,and cause pathological viral myocarditis and encephalitis damage.The result showed that the coinfection group was more seriously affected than the single-infection group,with worse clinical symptoms,increased viral replication,and obvious tissue pathological damage.This study provides a reference for further basic and clinical research into human enterovirus coinfection.

Background and aims:Hepatocellular carcinoma(HCC)is a malignant tumor with a high mortality rate,but there are still no effective treatments.The aim of this study was to investigate the anticancer po-tential of the combined use of brefeldin A(BFA)and tunicamycin(TM)in HepG2 cells,as well as the underlying mechanisms.Methods:HepG2 cells were treated with different concentrations of BFA(0.1-2.5 mg/L)and TM(1-5 mg/L)for 24 h.DMSO(0.1％,v/v)was used as a vehicle control.Cell viability and cell migration were measured using MTT assay and scratch wound assay,respectively.Apoptosis was detected using flow cytometry and acridine orange(AO)staining.The protein and mRNA levels of various factors involved in apoptosis(poly(ADP-ribose)polymerase-1(PARP-1),caspase-12,caspase-3,and stearoyl-CoA desaturase 1)and endoplasmic reticulum(ER)stress(binding immunoglobulin protein(BiP),protein kinase R-like endoplasmic reticulum kinase(PERK),p-PERK,phosphorylation of eukaryotic translation initiation factor 2alpha(p-eIF2α),activating transcription factor(ATF)4,and C/EBP homologous protein(CHOP))were measured using Western blotting and qRT-PCR,respectively.Results:Both BFA and TM alone significantly reduced the viability of HepG2 cells in a dose-dependent way.The co-incubation with TM(1 mg/L)further significantly reduced the viability of HepG2 cells treated with BFA(0.25 mg/L)alone(P＜0.05).BFA significantly increased the protein and mRNA levels of caspase-3 and PARP-1(P＜0.05)compared to control and DMSO-treated cells,indicating that BFA induced apoptosis in HepG2 cells by increasing the expression of caspase-3 and PARP-1.The induction of apoptosis by BFA could be further significantly enhanced by co-incubation with TM.In addition,BFA significantly increased the mRNA levels of BiP,PERK and ATF4(P＜0.05)compared to control and DMSO-treated cells.After co-incubation of BFA and TM,the protein levels of BiP,p-PERK,p-eIF2α and CHOP were significantly increased,indicating that TM could enhance BFA-induced ER stress in HepG2 cells through the PERK-eIF2α-ATF4-CHOP pathway.Conclusions:BFA could induce apoptosis and ER stress,and TM could enhance the ability of BFA to induce apoptosis and ER stress in HepG2 cells through the PERK-eIF2α-ATF4-CHOP pathway.The findings highlight the therapeutic potential of the combined use of BFA and TM in treating HCC.

Objective:To quantify and compare electromyographic(EMG)amplitude changes in the activation of the tibia-lis anterior muscle by 12 commonly used training methods in clinical rehabilitation training,and to identify the optimal activation method of the tibialis anterior muscle.Method:Sixteen healthy college students were participated in the study.Surface electromyography(sEMG)was used to quantify the activity level of the tibialis anterior muscle during 12 tests.Univariate variance was used to analyze the integrated electromyography(iEMG)and root mean square(RMS).Result:Among the 12 training methods targeting the activation of the tibialis anterior muscle,the maximum activation mode is T2"double feet standing(foam pad)ankle joint from zero to the maximum angle",fol-lowed by T1"double feet standing(floor),ankle joint from zero to the maximum angle"and T5"double feet standing(foam pad)ankle joint from zero to the maximum angle"foot dorsiflexion and invertion to the maxi-mum angle;The minimum activation was T12"the subject walked on the ground with a natural step",fol-lowed by T6"the subject jumped from a 30cm high step to the floor"and T7"the subject jumped from a 30cm high step to a foam pad".In RMS comparison,there was a statistical difference between the activation of T12,T6,T7,and T2(P＜0.05),and between the activation of T12,T6,and T1(P＜0.05).In iEMG com-parison,there were statistical differences(P＜0.05)between the activation of T12,T6,T7 and T2,T1,and T5(P＜0.05),and between the activation of T12,T6,and T3,with no significant statistical difference noted other comparisons.When comparing activation between male and female participants across the 12 tests,males generally had higher activation in the anterior tibial muscle than that in female,but no significant statistical difference was found.Conclusion:Increasing weight bearing,reducing support surface stability,increasing initial muscle initial length,and selecting difficulty training methods matching patient abilities can enhance the activation of anteri-or tibial muscle,while functional activities and balance strategies are not as effective in healthy population as other targeted tests.The training method for activating the tibialis anterior muscle is to stand on both feet(foam pad)with ankle dorsiflexion from a neutral position to maximum angle.The results of this study pro-vide information on the relative activation of the tibialis anterior muscle during specific movements,which can serve as evidence for prescribing exercise in clinical rehabilitation treatment and injury prevention.

Objective To establish an animal model of hand,foot,and mouth disease(HFMD)in Syrian hamsters coinfected with coxsackievirus A6(CVA6)and coxsackievirus B1(CVB1).Methods 42 Syrian hamsters were divided into a CVA6 infection group,CVB1 infection group,CVA6 and CVB1 coinfection group and control group.A HFMD model was established by nasal instillation of virus solution and phosphate-buffered saline.Clinical and physiological indicators and detoxification status were monitored and recorded for 15 d,and animals were selected on day 7(D7)after infection for histopathology and viral antigen and nucleic acid testing.Results Hamsters in the single-infection and coinfection groups showed clinical symptoms similar to human HFMD.White blood cell,neutrophil,and lymphocyte result were characteristic of viral infection.Both viral nucleic acids were detected in throat swabs,feces,blood,and tissues and both viruses were isolated from fecal samples.Pathological damage and positive co-localization of CVA6 and CVB1 viral antigen proteins and nucleic acids were found in brain and other tissues.Conclusions Nasal instillation of a CVA6 and CVB1 mixture can successfully coinfect Syrian hamsters,replicate herpes infection similar to human HFMD,and cause pathological viral myocarditis and encephalitis damage.The result showed that the coinfection group was more seriously affected than the single-infection group,with worse clinical symptoms,increased viral replication,and obvious tissue pathological damage.This study provides a reference for further basic and clinical research into human enterovirus coinfection.

Objective:To quantify and compare electromyographic(EMG)amplitude changes in the activation of the tibia-lis anterior muscle by 12 commonly used training methods in clinical rehabilitation training,and to identify the optimal activation method of the tibialis anterior muscle.Method:Sixteen healthy college students were participated in the study.Surface electromyography(sEMG)was used to quantify the activity level of the tibialis anterior muscle during 12 tests.Univariate variance was used to analyze the integrated electromyography(iEMG)and root mean square(RMS).Result:Among the 12 training methods targeting the activation of the tibialis anterior muscle,the maximum activation mode is T2"double feet standing(foam pad)ankle joint from zero to the maximum angle",fol-lowed by T1"double feet standing(floor),ankle joint from zero to the maximum angle"and T5"double feet standing(foam pad)ankle joint from zero to the maximum angle"foot dorsiflexion and invertion to the maxi-mum angle;The minimum activation was T12"the subject walked on the ground with a natural step",fol-lowed by T6"the subject jumped from a 30cm high step to the floor"and T7"the subject jumped from a 30cm high step to a foam pad".In RMS comparison,there was a statistical difference between the activation of T12,T6,T7,and T2(P＜0.05),and between the activation of T12,T6,and T1(P＜0.05).In iEMG com-parison,there were statistical differences(P＜0.05)between the activation of T12,T6,T7 and T2,T1,and T5(P＜0.05),and between the activation of T12,T6,and T3,with no significant statistical difference noted other comparisons.When comparing activation between male and female participants across the 12 tests,males generally had higher activation in the anterior tibial muscle than that in female,but no significant statistical difference was found.Conclusion:Increasing weight bearing,reducing support surface stability,increasing initial muscle initial length,and selecting difficulty training methods matching patient abilities can enhance the activation of anteri-or tibial muscle,while functional activities and balance strategies are not as effective in healthy population as other targeted tests.The training method for activating the tibialis anterior muscle is to stand on both feet(foam pad)with ankle dorsiflexion from a neutral position to maximum angle.The results of this study pro-vide information on the relative activation of the tibialis anterior muscle during specific movements,which can serve as evidence for prescribing exercise in clinical rehabilitation treatment and injury prevention.

Objective:To explore the value of Gestational Diabetes Mellitus Risk Score in monitoring hypoglycemia among neonates delivered by mothers with gestational diabetes mellitus (GDM) .Methods:From February 2018 to June 2020, 240 neonates delivered by mothers with GDM admitted to the Shanghai First Maternity and Infant Hospital affiliated to Tongji University were selected as the study subject. The patients were assessed and divided into groups by the Gestational Diabetes Mellitus Risk Score and the Supplementary Gestational Diabetes Mellitus Risk Score from 12 to 20 weeks. The low-risk group ( n=63) was defined as the score of Gestational Diabetes Mellitus Risk Score ≤ 20 and the score of Supplementary Gestational Diabetes Mellitus Risk Score<33 from 12 to 20 weeks. The group with the score of Gestational Diabetes Mellitus Risk Score>20 and the score of Supplementary Gestational Diabetes Mellitus Risk Score <33 from 12 to 20 weeks was classified as the medium-risk group ( n=49) . High-risk group ( n=128) was defined as the score of Gestational Diabetes Mellitus Risk Score >20 and the score of Supplementary Gestational Diabetes Mellitus Risk Score >33 from 12 to 20 weeks. Three groups of patients were tested for fasting blood glucose (FPG) . Within one hour after birth, neonates were tested for blood glucose using the glucose oxidase blood glucose meter and given early sucking. Within 72 hours after birth, continuous dynamic blood glucose monitoring methods were used to measure blood glucose in neonates. This study compared the blood glucose within one hour and at 6, 12, 24, 48, and 72 hours after birth and the incidence of hypoglycemia at 72 hours after birth among three groups. Results:Among the 240 neonates in the risk group, 197 experienced hypoglycemia, with an incidence rate of 82.08%. There were statistically significant differences in the incidence of hypoglycemia before treatment, one occurrence of hypoglycemia within 72 hours, and recurrent hypoglycemia among the three groups of neonates ( P<0.01) . There were statistically significant differences in blood glucose among the three groups of newborns within one hour, 6, 12, and 24 hours after birth ( P<0.05) . There was no statistically significant difference in blood glucose among the three groups at 48 hours and 72 hours after birth ( P>0.05) . Conclusions:Neonates with gestational diabetes will suffer from repeated hypoglycemia. The higher the risk level, the higher the incidence of hypoglycemia. It is important to assess the risk of neonates with GDM, monitor their blood glucose in real time and treat them accordingly.

【Objective】 To revise the Social Connectedness Scale-Revised （SCS-R） so as to explore the factor structure of SCS-R in the context of Chinese culture and test its reliability and validity. 【Methods】 Totally 900 college students were asked to complete the Chinese version of the Social Connectedness， and 850 valid questionnaires were collected. In addition， the Distress Disclosure scale （DDI） and UCLA Loneliness Scale were used as the criteria. One month later， 100 students were randomly selected for retest. 【Results】 ① The results of exploratory factor analysis showed that one factor with more than one eigenvalue was selected， and the cumulative variance explained was 62.44%. ② Confirmatory factor analysis showed that the data fit well （χ²=33.438， df=14， χ²/df=2.388， RMSEA=0.04， CFI=0.996， TLI=0.991）. ③ The Chinese version of the social connectedness scale is a one-dimensional scale. Its internal consistency coefficient is 0.916 and test-retest reliability is 0.845. ④ Social connectedness is significantly positively correlated with distress disclosure and negatively correlated with loneliness， indicating that the scale has good criterion-related validity. 【Conclusion】 The Chinese version of the social connectedness scale has satisfactory reliability and validity， and has cross-cultural adaptability.

Objective:To evaluate the clinical efficacy and safety of fractional CO 2 laser for the treatment of vaginal relaxation syndrome. Methods:Twenty-one females with vaginal relaxation syndrome receiced between December 2016 and January 2017 in our department were treated for 3 times at 1 month interval with femilift fractional CO 2 laser. Vaginal palpation, vaginal squeeze pressure test, vaginal ultrasound, pelvic floor MRI, and Incontinence Questionnaire Short-Form (ICIQ-SF) assessments were performed at baseline and at post-treatment. Results:Significant improvement in female sexual dysfunction and stress urinary incontinence was especially seen based on patient＇s feelings after treatments. A noted progressive differences were observed in FSFI and vaginal pressure test, which were much higher after treatment than before treatment ( P<0.01). Moreover, in 13 cases suffering from mild to moderate stress urinary incontinence (SUI), fractional CO 2 laser treatment was associated with a significant improvement of ICIQ-SF scores ( P<0.01). All subjects successfully completed the study with no adverse events such as infections, bleeding and pain, etc. Conclusions:Fractional CO 2 laser treatment is an effective treatment for female sexual dysfunction and stress urinary incontinence caused by vaginal relaxation syndrome.

OBJECTIVETo detect potential mutation of TCOF1 gene in a Chinese family affected with Treacher-Collins syndrome.

METHODSClinical data of the patient was collected. The analysis included history taking, clinical examination and genetic testing. All coding regions of the TCOF1 gene were subjected to PCR amplification and Sanger sequencing.

RESULTSA novel mutation c.2261ins G (p.E95X) of the TCOF1 gene was discovered in the patient. The same mutation was not found in his parents and 100 healthy controls.

CONCLUSIONThe c.2261insG (p.E95X) mutation of the TCOF1 gene probably underlies the disease in the patient. Genetic testing can facilitate diagnosis and genetic counseling for families affected with TCS.

Objective:To investigate and compare the effects of water extracts of Whitmania pigra Whitman and Hirudinaria ma-nillensis Lesson on the angiogenesis of Tg (kdrl:mCherry) zebrafish. Methods:The zebrafish embryos 6-8 hours after fertilization (6-8hpf) were transferred to the culture medium containing Whitmania pigra Whitman or Hirudinaria manillensis Lesson extract at different concentrations, and the culture medium containing the drugs was replaced every 24 h. And then, at 72 hpf, the larvalmorphology and intersegmental vessels were observed under a microscope. The hatchability of 48-and 72-hpf embryos, and the number of intersegmen-tal vessels and the heart rate of 72-hpf juveniles were measured. Results:Compared with the control group, when the concentration of Whitmania pigra Whitmanis was higher than 30μg· ml-1 , and the concentration of Hirudinaria manillensis Lesson was higher than 20μg· ml-1, the number of intersegmental vessels was significantly reduced (P<0. 01). Compared with the control group, at 48 hpf, when the concentration of the drug groups was higher than 40 μg· ml-1 , the hatchability of the two groups significantly decreased ( P<0. 01);at 72 hpf, the hatchability of Whitmania pigra Whitman decreased significantly at the concentration of 100 μg·ml-1 (P<0.01), while the hatchability of Hirudinaria manillensis Lesson decreased significantly at the concentration of 80 μg· ml-1(P <0. 01). There was no obvious yolk sac edema, pericardial edema and spine curvature in the two groups. The heart rate decreased sig-nificantly (P<0. 01), while was still within the normal range. Conclusion:Both Whitmania pigra Whitman and Hirudinaria manillen-sis Lesson have notable anti-angiogenic activity, and the anti-angiogenesis activity of Hirudinaria manillensis Lesson is stronger. They both have effects on the development of zebrafish embryos, while the toxicity is not obvious.