Objective:To construct a recombinant herpes simplex virus 2 (HSV-2) expressing enhanced green fluorescent protein (EGFP) using clustered, regularly interspaced, short palindromic repeat/CRISPR-associated nuclease 9 (CRISPR/Cas9) technology.Methods:Four strategies for inserting exogenous EGFP gene into HSV-2 genome using CRISPR/Cas9 technology were designed: (1) conventional homology-directed repair: circular two homology arm donor-mediated gene knock-in; (2) linearized single homology arm donor-mediated gene knock-in; (3) homology-independent targeted integration; (4) conventional homology-directed repair-mediated by cell lines stably expressing Cas9 and sgRNA.Results:The recombinant virus HSV-2-EGFP was successfully constructed based on the second, the third and the fourth strategies. The second strategy was the most efficient, followed by the third and the fourth strategies. The purified recombinant virus could stably express green fluorescent protein in seven passages and shared similar growth characteristics in Vero cells to the parental virus.Conclusions:Linearized single homology arm donor could increase the efficiency of gene knock-in, and cell lines stably expressing Cas9 and sgRNA could increase the efficiency of gene knock-in mediated by homology-directed repair.

Objective:To develop a nomogram model for screening of type 2 diabetes mellitus in a community population.Methods:From October to December, 2020, 6 028 community residents who participated in the " national health physical examination" in Karamay community with complete physical examination data and met the inclusion and exclusion criteria were selected. The physical examination data included medical history, physical examination, laboratory, and ultrasound reports. Random segmentation sampling was used to divide the population into modeling and validation cohorts, and LASSO regression analysis was used to screen for independent factors associated with diabetes diagnosis. The independent influencing factors were furthor incorporated into the multi-factor logistic regression, and the RMS software package was used to construct the column chart. The area under receiver operating characteristic(ROC) curve was used to measure the differentiation of the model. The calibration curve can directly reflect the calibration degree of the model.Results:In the modeling group, multivariate logistic regression analysis showed that age, gender(female), history of hypertension, history of hyperlipidemia, HbA 1C, urinary microalbumin, homeostasis model assessment for insulin resistance, and triglycerides and glucose index were independently associated with diabetes. OR were 1.053(95% CI 1.038-1.069), 0.681(95% CI 0.512-0.906), 1.802(95% CI 1.227-2.626), 1.789(95% CI 1.303-2.448), 10.973(95% CI 8.318-14.745), 1.002(95% CI 1.001-1.004), 2.914(95% CI 2.248-3.799), and 2.673(95% CI 2.03-3.536), respectively. The areas under ROC curves of the training set and the validation set were 0.945 and 0.955, respectively. The optimal critical value in the ROC curve was 0.178(sensitivity 0.930, specificity 0.839) in the training set and 0.201(sensitivity 0.945, specificity 0.848) in the validation set. Conclusion:The screening model of type 2 diabetes developed in this study has good accuracy, which can be used as a screening tool for high-risk population of type 2 diabetes.

Objective To investigate the community structure and spatial distribution of wild myomorpha in the plague foci of Lianghe County,Yunnan Province.Methods From December 2015 to October 2016,in Mangdong,Hexi,Nangsong and other places in Lianghe County,forest landscape samples were randomly selected in different altitude gradients (1 000 ～,1 200 ～,1 400 ～,≥ 1 600 m).Peanut was used as a bait for trapping myomorpha,and the community structure of myomorpha was analyzed.Results A total of 663 myomorpha were collected and the density was 7.97％ (663/8 320),which were divided into 16 species,11 genus,5 families,and 3 orders.Rattus tanezumi (30.62％,203/663) was the dominant species.In the 1 000-＜ 1 200 m altitude gradient,the richness of myomorpha was the highest (13 kinds).In the 1 400-＜ 1 600 m altitude gradient,the index of diversity (2.094 2) was the highest.With the increase of altitude,the ecological dominance were increased,and the values were 0.672 2,0.774 2,0.842 8,and 0.882 4,respectively,whereas the evenness presented a decreasing trend,and the values were 0.269 2,0.197 1,0.142 8,and 0.162 0.Conclusions The Rattus tanezumi has presented a single dominant distribution at different elevation and the number and the species of the wild rodent with superior position have increased with the altitude increased in Lianghe focus.With the increase of altitude,the diversity and the evenness have increased,whereas the richness and the dominance have decreased.

Objective:To investigate the effects of genomic location of a foreign gene in Shanghai-191 strain measles virus (MV) vector on gene expression and virus replication.Methods:The nucleotide sequence encoding S1 protein of SARS-CoV-2 was inserted at different positions in MV antigenome (the upstream of the N gene, between P and M genes, between H and L genes), and co-transfected into 293T cells with helper plasmids coding T7 RNA polymerase and N, P, and L proteins, respectively. The transfected cells were lysed and the supernatants were used to infected Vero cells to harvest recombinant viruses. S1 proteins expressed by the recombinant viruses were identified by RT-PCR, indirect immunofluorescence assay, Western blot and ELISA. Growth kinetics of the recombinant viruses were analyzed.Results:Recombinant viruses were failed to be rescued when the S1 protein-coding sequence was cloned into the upstream of N gene. Two recombinant viruses, MV-M-S1 and MV-L-S1, were successfully rescued when cloning the S1 protein-coding sequence into the intergenic region between P and M genes, or H and L genes, and could express S1 protein. MV-M-S1 expressed more S1 protein than MV-L-S1, but the titer of MV-M-S1 was lower.Conclusions:Inserting a foreign gene at different positions in the MV genome might have different effects on gene expression and virus replication. This study provided reference for the subsequent construction of MV vector.

Objective To investigate the geographic landscape features in Yulong plague natural focus.Methods Four altitudinal gradients (2 400-,2 600-,2 800-,≥3 000 m) were selected as study objects in Yulong plague natural focus from December 2015 to November 2016 to collect and analyze geographic landscape factors (topography and terrain,climate,water areas,vegetation and soil) based on the literatures about this plague focus.Results Yulong plague natural focus was an area with mid-mountain and subalpine topography in northwestern Yunnan Province,and its terrain had the character of lower in the southeast and higher in the northwest.This area belonged to plateau warm temperate monsoon climate of low latitude with an average month temperature 13.6 ℃ from 2005-2016,month air relative humidity averaged 59.4％,and rainfall averaged 884.5 mm a year,which concentrated mostly from June to September.About 98.1％ of the water area in Yulong County was controlled by Chin-sha River.The vegetation and soil were characterized by vertical distribution along with altitude gradients obviously.Conclusion The unique geographical landscape of the plague natural focus in Yulong County is one of important factors affecting the occurrence and epidemic of the plague.

Objective To compare and analyze the soil eight metal elements contents of rat holes of three plague foci in Yunnan Province.Methods The soils were collected from the rat holes in Yulong,Jianchuan,and Lianghe plague foci.Calcium (Ca),iron (Fe),zinc (Zn),chromium (Cr),lead (Pb),manganese (Mn),cadmium (Cd),and copper (Cu) were quantified with flame atomic absorption spectrometry (FAAS).Data input,calculation and collation were conducted through Excel 2007,and statistical software SPSS 20.0 was used to carried out statistical analysis through rank sum test.Results The medians of Ca content in the rat hole soil of Yulong County,Jianchuan County,and Lianghe County,respectively,were 85.40,308.56,and 0.00 mg/kg;the medians of Fe were 2 569.07,1 950.16,and 975.72 mg/kg;the medians of Zn were 5.64,4.02,and 2.04 mg/kg;the medians of Cr were 1.09,3.06,and 0.71 mg/kg;the medians of Pb were 3.61,29.56,and 22.80 mg/kg;the medians of Mn were 223.01,212.97,and 80.43 mg/kg;the medians of Cd were 0.00,2.02,and 2.94 mg/kg;and the medians of Cu were 1.92,1.08,and 0.51 mg/kg.The contents of eight metal elements were statistically different in the soil samples between Yulong,Jianchuan,and Lianghe plague foci (H =80.538,127.687,202.583,185.359,124.747,150.057,91.346,228.546,P ＜ 0.05).Conclusions The content of Fe in soil of the three plague foci in Yunnan Province is abundant.The contents of metal elements of Yulong County and Jianchuan County are similar.

Objective:To evaluate clinical efficacy and safety of topical compound oleum lithospermi in the treatment of mild to moderate diaper dermatitis.Methods:A multicenter, randomized, positive-drug parallel-controlled clinical trial was conducted in 19 hospitals from July 2019 to August 2020. Children aged 0 - 12 months with mild to moderate diaper dermatitis were enrolled and randomly divided into 2 groups using a random number table: test group topically treated with compound oleum lithospermi, and control group topically treated with zinc oxide cream. The treatment was carried out 6 - 8 times a day for 7 days. Visits were scheduled on days 0 and 7, and total response rate and clinical healing time were evaluated. Changes in the dermatitis family impact (DFI) score were compared between the test group and control group, and adverse events were recorded. Statistical analysis was carried out by using independent-sample t test for normally distributed continuous data, Wilcoxon rank sum test for non-normally distributed continuous data, and chi-square test or Fisher′s exact test for unordered categorical data; survival curves were drawn, and log-rank test was used for comparisons between two groups. Results:A total of 343 children with diaper dermatitis were enrolled in this study. Among them, 31 children violated the protocol, so 312 were included in the per protocol set, including 157 in the test group and 155 in the control group, and all completed the visits on days 0 and 7. The total response rate was significantly higher in the test group (87.26%, 137/157) than in the control group (78.71%, 122/155; χ2 = 4.04, P = 0.044) . The clinical healing time was significantly shorter in the test group (5.33 days) than in the control group (6.13 days; χ2 = 4.67, P = 0.025) . After 7-day treatment, the DFI score significantly decreased in both the 2 groups compared with that before the treatment, but there was no significant difference in the DFI score between the 2 groups (test group: 4.02 ± 6.96, control group: 3.58 ± 5.90, Z = -0.39, P = 0.686) . The incidence of adverse events was 2.92% (5/171) and 5.45% (9/165) in the test group and control group respectively, and there was no significant difference between the 2 groups ( χ2 = 0.03, P = 0.865) . Conclusion:Compound oleum lithospermi can markedly reduce the clinical severity of diaper dermatitis, improve the total response rate, shorten the clinical treatment period, and improve the quality of life of children′s families with a favorable safety profile.

OBJECTIVE To monitor the occurrence of tenofovir disoproxil fumarate （TDF）-induced kidney injury and investigate the risk factors， and provide reference for rational use of TDF in clinic. METHODS The information of inpatients with hepatitis B was collected by China Hospital Pharmacovigilance System （CHPS） from the Second People’s Hospital of Lanzhou during Jan. 1st， 2019 to Dec. 31st 2021. The search criteria were set according to kidney injury criteria， and suspected TDF- induced kidney injury cases were actively monitored； then the clinical pharmacist confirmed the positive patients with TDF-induced kidney injury one by one and calculated the incidence of TDF-induced renal injury； the risk factors for TDF-induced kidney injury in real world were explored by collecting and analyzing the correlation of basic data of patients， main indexes of liver and kidney function， complications and combined use of drugs with TDF-induced renal indexes. RESULTS Totally 1 226 inpatients with hepatitis B using TDF were included. Through active monitoring of CHPS， 160 suspected patients with TDF-induced kidney injury were found， and 64 positive patients were finally confirmed manually. The incidence of TDF-induced kidney injury was 5.22%. Compared with pre-medication， the levels of serum creatinine and cystatin C， the proportion of patients with urinary protein 2+ and above were increased significantly after medication （P＜0.001）， glomerular filtration rate and blood phosphorus level were reduced significantly （P＜0.001） and other indicators had no statistical difference. Treatment time for more than 36 months， disease progresses to decompensated cirrhosis， and concomitant use of more than 10 kinds of drugs were significantly correlated with TDF- related kidney injury （P＜0.05 or P＜0.012 5）. CONCLUSIONS The active monitoring scheme of TDF-induced kidney injury established by CHPS has the characteristics of time-saving， labor-saving and high efficiency； based on real-world evidence， it is imperative to strengthen monitoring kidney function of patients when using TDF， especially when the patient has been on medication for a long time， in decompensated cirrhosis and combination of multiple drugs， and thus， we can identify earlier and avoid adverse effects in high-risk patientseffectively.

Compared to conventional cancer treatment, combination therapy based on well-designed nanoscale platforms may offer an opportunity to eliminate tumors and reduce recurrence and metastasis. In this study, we prepared multifunctional microspheres loading I-labeled hollow copper sulfide nanoparticles and paclitaxel (I-HCuSNPs-MS-PTX) for imaging and therapeutics of W256/B breast tumors in rats. F-fluordeoxyglucose (F-FDG) positron emission tomography/computed tomography (PET/CT) imaging detected that the expansion of the tumor volume was delayed (<0.05) following intra-tumoral (i.t.) injection with I-HCuSNPs-MS-PTX plus near-infrared (NIR) irradiation. The immunohistochemical analysis further confirmed the anti-tumor effect. The single photon emission computed tomography (SPECT)/photoacoustic imaging mediated by I-HCuSNPs-MS-PTX demonstrated that microspheres were mainly distributed in the tumors with a relatively low distribution in other organs. Our results revealed that I-HCuSNPs-MS-PTX offered combined photothermal, chemo- and radio-therapies, eliminating tumors at a relatively low dose, as well as allowing SPECT/CT and photoacoustic imaging monitoring of distribution of the injected agents non-invasively. The copper sulfide-loaded microspheres, I-HCuSNPs-MS-PTX, can serve as a versatile theranostic agent in an orthotopic breast cancer model.

Objective: To investigate whether topical mucopolysaccharide polysulfate (MPS) cream can reduce the incidence of eczema and skin atrophy in patients with moderate- or low-risk infantile hemangioma after the treatment with topical beta-blockers or 595-nm pulsed dye laser (PDL) , and to analyze factors influencing the occurrence of eczema and skin atrophy. Methods: A total of 722 patients aged 0- 1 years with moderate- or low-risk infantile hemangioma were enrolled from 5 Children′s Hospitals in China. According to the disease condition and therapy acceptability, these patients were divided into 6 groups to be treated with topical beta-blockers and MPS cream (blocker+ MPS group) , topical beta-blockers (blocker group) , 595-nm PDL and topical MPS cream (PDL+ MPS group) , 595-nm PDL (PDL group) , 595-nm PDL combined with topical beta-blockers and MPS cream (PDL+ blocker+ MPS group) , and 595-nm PDL and topical beta-blockers (PDL+ blocker group) , respectively. All the externally applied agents were applied twice a day, and PDL was performed once every 4 weeks. Efficacy and adverse reactions were evaluated after 3-month treatment. Univariate and multivariate Logistic regression analyses were carried out to analyze factors influencing the incidence of eczema and skin atrophy in patients with infantile hemangioma after treatment, and chi-square test was carried out to compare efficacy among the groups. Results: After 3-month treatment, multivariate Logistic regression analysis for comparing the blocker+ MPS group with blocker group showed that the risk factor for eczema on the surface of hemangiomas was no topical treatment with MPS cream (P= 0.007, OR= 3.887, 95% CI: 1.439-10.493) , while no correlations were observed between the occurrence of skin atrophy on the surface of hemangiomas and analyzed factors. Multivariate Logistic regression analysis for comparing the PDL+MPS group with PDL group showed that no topical treatment with MPS cream (P < 0.001, OR= 7.402, 95% CI: 2.604-21.042) and northern areas (P < 0.001, OR= 67.048, 95% CI: 7.977-563.518) were risk factors for eczema on the surface of hemangiomas, and risk factors for skin atrophy on the surface of hemangiomas included no topical treatment with MPS cream (P = 0.001, OR = 9.371, 95 CI: 2.590 - 33.900) and abundant blood supply of hemangiomas (P = 0.036, OR = 2.971, 95% CI: 1.075-8.208) . Multivariate Logistic regression analysis for comparing the PDL + blocker+ MPS group with PDL+ blocker group showed that risk factors for eczema on the surface of hemangiomas were no topical treatment with MPS cream (P= 0.005, OR= 3.426, 95% CI: 1.446-8.119) and northern areas (P < 0.001, OR= 31.704, 95% CI: 6.924-145.158) , and risk factors for skin atrophy on the surface of hemangiomas included no topical treatment with MPS cream (P < 0.001, OR= 6.011, 95% CI: 2.558-14.126) and southern areas (P= 0.022, OR= 3.021, 95% CI: 1.177-7.753) . After 3-month treatment, the response rate was significantly higher in the PDL group than in the PDL+ MPS group (χ²= 4.531, P= 0.033) , and significantly higher in the blocker group than in the blocker+ MPS group (χ²= 4.344, P= 0.037) . There were no significant differences in the response rate or cure rate among the other groups (all P > 0.05) . Conclusion During the treatment of moderate- or low-risk infantile hemangioma with topical beta-blockers or 595-nm PDL, the combination with topical MPS cream can reduce the occurrence of eczema and skin atrophy without affecting the therapeutic effect.