Objective:To construct a full-length infectious clone of enterovirus D68（EV-D68）expressing enhanced green fluorescent protein（EGFP）by reverse genetics in order to provide an efficient tool for studying the biological characteristics and screening antiviral drugs for EV-D68.Methods:Gene synthesis and overlap PCR techniques were used to construct the full-length clone plasmid pUC57-EV-D68 of EV-D68. The full-length viral sequence was then transferred into the pCAGGS plasmid to obtain the pCAGGS-EV-D68 plasmid. The EGFP gene was amplified and inserted into the pCAGGS-EV-D68 plasmid to construct the pCAGGS-EGFP-EV-D68 plasmid. Then，the two constructed plasmids were transfected into human rhabdomyosarcoma（RD）cells to rescue recombinant viruses RV-EV-D68 and RV-EGFP-EV-D68. The rescued viruses were identified using PCR，Western blot，and immunofluorescence techniques. The antiviral effect of doxycycline was evaluated using the rescued RV-EGFP-EV-D68. Statistical analysis was performed using the two independent samples t-test. Results:The recombinant virus RV-EGFP-EV-D68 capable of expressing EGFP was successfully rescued. Even after 15 serial passages，the virus retained EGFP expression with no significant difference in viral titers compared to the parental virus，indicating its stable passage in RD cells. Besides，the rescued strains exhibited similar replication characteristics to the parental virus. While at 24 and 36 h after infection，the titers of the rescued strains were significantly lower than that of the parental strain（both P<0.05）. This study demonstrated that doxycycline significantly reduced the fluorescence intensity of RV-EGFP-EV-D68-infected RD cells（ P<0.01）. Meanwhile，a negative correlation was observed between the doxycycline concentration and the fluorescence intensity，indicating that the rescued virus could be used for antiviral drug evaluation. Conclusions:This study successfully constructs an infectious clone of EV-D68 expressing EGFP. The rescued recombinant virus RV-EGFP-EV-D68 has been verified to be applicable for the evaluation of antiviral drugs.

Objective:To construct a full-length infectious clone of enterovirus D68（EV-D68）expressing enhanced green fluorescent protein（EGFP）by reverse genetics in order to provide an efficient tool for studying the biological characteristics and screening antiviral drugs for EV-D68.Methods:Gene synthesis and overlap PCR techniques were used to construct the full-length clone plasmid pUC57-EV-D68 of EV-D68. The full-length viral sequence was then transferred into the pCAGGS plasmid to obtain the pCAGGS-EV-D68 plasmid. The EGFP gene was amplified and inserted into the pCAGGS-EV-D68 plasmid to construct the pCAGGS-EGFP-EV-D68 plasmid. Then，the two constructed plasmids were transfected into human rhabdomyosarcoma（RD）cells to rescue recombinant viruses RV-EV-D68 and RV-EGFP-EV-D68. The rescued viruses were identified using PCR，Western blot，and immunofluorescence techniques. The antiviral effect of doxycycline was evaluated using the rescued RV-EGFP-EV-D68. Statistical analysis was performed using the two independent samples t-test. Results:The recombinant virus RV-EGFP-EV-D68 capable of expressing EGFP was successfully rescued. Even after 15 serial passages，the virus retained EGFP expression with no significant difference in viral titers compared to the parental virus，indicating its stable passage in RD cells. Besides，the rescued strains exhibited similar replication characteristics to the parental virus. While at 24 and 36 h after infection，the titers of the rescued strains were significantly lower than that of the parental strain（both P<0.05）. This study demonstrated that doxycycline significantly reduced the fluorescence intensity of RV-EGFP-EV-D68-infected RD cells（ P<0.01）. Meanwhile，a negative correlation was observed between the doxycycline concentration and the fluorescence intensity，indicating that the rescued virus could be used for antiviral drug evaluation. Conclusions:This study successfully constructs an infectious clone of EV-D68 expressing EGFP. The rescued recombinant virus RV-EGFP-EV-D68 has been verified to be applicable for the evaluation of antiviral drugs.

To promote the development of the prescription-based processing of Chinese herbal pieces,the China Association of Chinese Medicine published the social organization standard of the Standard for Prescription-based Processing of Chinese Herbal Pieces(T/CACM 1367-2021)in June 2021.The standard was led by the First Affiliated Hospital of Henan University of Chinese Medicine and Jiangsu Province Hospital of Chinese Medicine.It was jointly drafted by 28 Traditional Chinese medical institutions across the country.This paper introduced the standards in detail to promote the implementation and propel the inheritance and innovation of the processing of Chinese herbal pieces.

Objective To observe the value of grey-level histogram analysis based on T2WI for differentiating consistency of meningioma.Methods Data of 109 patients with meningioma were retrospectively analyzed.The patients were divided into hard group(n=71)and soft group(n=38)according to the consistency of tumors.Tumor ROI was outlined on axial T2WI showing the largest tumor section,gray levels were extracted and histogram analysis was performed.The value of each histogram parameter were compared between groups.Then receiver operating characteristic curve was drawn,and the area under the curve(AUC)was calculated to evaluate the efficiency for differentiating soft and hard meningioma.Results P1,P10,P50,P90,P99 and the mean grey levels on T2WI in soft group were all higher than those in hard group(all P＜0.05),while the variance,the kurtosis and the skewness were not significantly different between groups(all P＞0.05).The differentiating efficiency of P1,P10,P50,P90,P99 and the mean grey levels on T2WI were all fine,with AUC of 0.774 to 0.833,and no significant difference was found(all P＞0.05).Conclusion Parameters of grey-level histogram analysis such as P1,P10,Ps0,P90,P99 and the mean values based on T2WI were all valuable for differentiating soft and hard meningioma.

ObjectiveTo explore the effect of nutrition combined with exercise intervention on stroke patients with sarcopenia. MethodsFrom January to June, 2022, 60 stroke patients with sarcopenia were randomly divided into control group (n = 15), nutrition group (n = 15), exercise group (n = 15) and combined group (n = 15). All the groups received routine rehabilitation training, while the nutrition group received nutrition intervention, the exercise group received exercise intervention, and the combined group received both the nutrition and exercise intervention, for four weeks. Before and after intervention, the muscle index was measured with bioelectrical impedance analysis, gripping strength of the healthy and the affected side was measured with gripping strength meter, and the patients were assessed with modified Barthel Index (MBI) and Berg Balance Scale (BBS). ResultsFour cases in the control group, two in the nutrition group, one in the exercise group, and three in the combined group dropped down. The muscle index, gripping strength, and the scores of MBI and BBS improved in all the groups after intervention (|t| > 3.004, P < 0.05), while all improved more in the combined group than in the other three groups (P < 0.05), and the grip strength of the healthy side was more in the exercise group than in the nutrition group (P < 0.05). ConclusionNutrition or exercise intervention alone can improve the muscle quality, grip strength, activities of daily living and balance of stroke patients with sarcopenia, while the combination is more effective.