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.

Objective:To explore the effect of external application of Traditional Chinese Medicine on preventing peripherally inserted central catheter (PICC) associated phlebitis in tumor patients by network Meta-analysis.Methods:Randomized controlled trials on the prevention of PICC associated phlebitis in tumor patients with external application of Traditional Chinese Medicine were searched in PubMed, Web of Science, Embase, Cochrane Library, CINAHL, China National Knowledge Infrastructure, WanFang Database, and China Biomedical Medicine disc. The search period was from the establishment of the database to July 1, 2022. Stata 17.0 software was used for network Meta-analysis.Results:A total of 15 articles were included, including 1 160 patients and 13 types of Traditional Chinese Medicine. Probability ranking according to the surface under the cumulative ranking curve showed that the top four traditional Chinese medicines in terms of effectiveness in preventing PICC associated phlebitis in tumor patients were Rhizoma Corydalis Mixture, Liuhe Dan, Qingfu Ointment, and Ruhuangbing Ointment.Conclusions:The external application of Rhizoma Corydalis Mixture, Liuhe Dan, Qingfu Ointment, and Ruhuangbing Ointment can effectively prevent the occurrence of PICC associated phlebitis in tumor patients. Although Rhizoma Corydalis Mixture has the best effect in preventing phlebitis, due to the limited sample size involved in the analysis and insufficient accuracy of the combined effects, its effectiveness in preventing phlebitis can be further verified in the future.

Idiopathic pulmonary fibrosis (IPF) is a progressive disease with unknown etiology and limited therapeutic options. Activation of fibroblasts is a prominent feature of pulmonary fibrosis. Here we report that lncRNA DACH1 (dachshund homolog 1) is downregulated in the lungs of IPF patients and in an experimental mouse model of lung fibrosis. LncDACH1 knockout mice develop spontaneous pulmonary fibrosis, whereas overexpression of LncDACH1 attenuated TGF-β1-induced aberrant activation, collagen deposition and differentiation of mouse lung fibroblasts. Similarly, forced expression of LncDACH1 not only prevented bleomycin (BLM)-induced lung fibrosis, but also reversed established lung fibrosis in a BLM model. Mechanistically, LncDACH1 binding to the serine/arginine-rich splicing factor 1 (SRSF1) protein decreases its activity and inhibits the accumulation of Ctnnb1. Enhanced expression of SRSF1 blocked the anti-fibrotic effect of LncDACH1 in lung fibroblasts. Furthermore, loss of LncDACH1 promoted proliferation, differentiation, and extracellular matrix (ECM) deposition in mouse lung fibroblasts, whereas such effects were abolished by silencing of Ctnnb1. In addition, a conserved fragment of LncDACH1 alleviated hyperproliferation, ECM deposition and differentiation of MRC-5 cells driven by TGF-β1. Collectively, LncDACH1 inhibits lung fibrosis by interacting with SRSF1 to suppress CTNNB1 accumulation, suggesting that LncDACH1 might be a potential therapeutic target for pulmonary fibrosis.

Objective:To construct the forewarning model of drug shortage in Shanxi province, so as to realize the early prediction of drug shortage.Methods:According to the drug shortage of data of each monitoring station in Shanxi province, from the four dimensions of drug factors, policy attributes, supplier factors and demander factors, 14 factors were selected for grey correlation analysis, for example whether they were urgent drugs for women and children, shortage types, etc. The main factors affecting the degree of drug shortage were selected, and the early warning analysis model based on two-step cluster analysis method was established.Results:A total of six factors with the highest correlation with the degree of drug shortage were determined in this study, in order: whether they were urgent drugs for women and children, shortage types, low-price drugs, the number of moving annual total monitoring enterprises in sale in the current month, shortage reasons, and whether they were basic drugs. Based on the two-step cluster analysis, a model of drug shortage forewarning in Shanxi Province was established, which was divided into four optimal clusters and the warning level was determined.Conclusions:This study establishes the early warning model of drug shortage in Shanxi province, which is helpful to find the risk of drug shortage as soon as possible, objectively predict the risk level, and assist the drug regulatory department to realize layered implementation and collaborative response.

Objective:To provide more options for preoperative localization diagnosis in patients with primary hyperparathyroidism (PHPT), the diagnostic efficacy of parathyroid 4-dimensional computed tomography (4D-CT) in patients with PHPT was evaluated.Methods:This was a single-center retrospective study including 57 patients with surgical proved PHPT. All of the patients underwent 4D-CT, 99Tc m -sestamibi parathyroid imaging (MIBI), and ultrasonography (US) preoperatively. The reference standard for correct localization was based on operation reports and pathology confirmation. The patients were grouped according to the preoperative serum calcium levels, tumor diameter, or ectopic lesions (yes/no), respectively. The sensitivity, specificity, positive predictive value, negative predictive value and area under the curve (AUC) of 4D-CT, MIBI and US, alone or in combination, were analyzed in total and each subgroup patients. Results:Fifty-seven patients (39 women, 18 men; mean age of 56.5 years) were evaluated, including four cases with multi-gland disease and thirteen cases with ectopic parathyroid lesions. In all the patients, similar diagnostic efficacy was found in 4D-CT (AUC: 0.943) and MIBI (AUC: 0.927), both of which were higher than that of US (AUC: 0.847) ( P = 0.01 for 4D-CT vs. US; P = 0.04 for MIBI vs. US). In a subset analysis for ectopic quadrants, the diagnostic efficacy of 4D-CT was significantly higher than that of MIBI ( P = 0.04) or US ( P = 0.01), with the sensitivity of 100%, 69.2%, and 61.5%, and AUC of 0.989, 0.846, and 0.808 for 4D-CT, MIBI and US, respectively. Conclusions:4D-CT has similar diagnostic efficacy for preoperative localization to MIBI in patients with PHPT, and it is superior to MIBI and US in identifying the ectopic parathyroid gland. 4D-CT can be recommended as an alternative preoperative localization method, especially when parathyroid lesions could not be precisely located by US and MIBI.

Objective:To conduct periodic revalidation of the 15 items and 43 terms autoverification rules of blood analysis after 1 year of application, analyze the application suitability and make the rules improved.Methods:Track the results of 528 010 blood analysis samples of our hospital from August 1, 2019 to January 31, 2020, and analyze the pass rate and interception rate of autoverification; 600 specimens in total were selected randomly for microscope examination, including 300 specimens which touched autoverification rules (1 012 items of autoverification rules) and were intercepted by autoverification and 300 specimens which untouched autoverification rules and were released by autoverification. The abnormal characteristics and unacceptable Delta check of the specimens also need to be concerned at the same time.The false negative rate and false positive rate, true negative rate, true positive rate and pass correct rate of autoverification were verified and compared with the rate of the second phase verification when the autoverification rule was established. The false negative rate, false positive rate, true negative rate and true positive rate of the Delta check rule which 54 716 specimens touched were calculated and compared with the second phase verification rate when the autoverification rule was established.The results of microscopic examination were used as the gold standard for the calculation of the rates, and P<0.05 was considered as a significant difference. The false positive and true positive of 1 012 autoverification rules were analyzed item by item.The false positive and true positive of 108 specimens which touched blast cell autoverification rule were analyzed terms by terms. The mean TAT and median TAT of 528 010 specimens and 193 750 outpatient specimens were calculated respectively, and the report percentages of 528 010 samples that TAT<30, 30-60　and>60 min were calculated respectively. Analyze and evaluate the application suitability of autoverification rules to juge whether they meet the needs of doctors and laboratory. The design process and the rules and application process of autoverification were optimized and improved.Results:The autoverification pass rate was 63.06% (332 971/528 010), the interception rate was 36.94% (195 039/528 010). The false negative rate was 1.00% (1/600), the false positive rate was 12.67% (76/600), the true negative rate was 49% (294/600), the true positive rate was 37.33% (224/600), and the correct rate was 98% (294/300). The pass rate, true negative rate, true positive rate and correct rate of the periodic reverification group were higher than the second phase verification group, the false negative rate and false positive rate were lower than that the second phase verification group. The false negative rate and true positive rate of the Delta check of periodic verification group were lower than that the second phase verification group, the false positive rate and true negative rate were higher than the second phase verification group, there were significant differences in the comparition results. The mean TAT of 528 010 specimens was25 min, and the median TAT was 22 min. The mean TAT of 193 750 outpatient specimens was 23 min, and the median TAT was 20 min. The report percentages of 528 010 samples that TAT<30 min, 30 min-60 min and>60 min were 83.30% (439 819/528 010), 8.00% (42 250/528 010) and 8.70% (45 941/528 010), respectively.Conclusion:The results of periodic revalidation of autoverification after 1 years application show that the 15 items and 43 terms autoverification rules of blood analysis could meet requirements about the accuracy and efficiency of the laboratory, and have a good suitability for application.

Objective　To establish Huashan diagnostic criteria and clinical classification system for Hirayama disease.Methods　Retrospective analysis 359 cases of puberty onset,upper extremity muscle atrophy as main clinical manifestations,and complete clinical data from September 2007 to August 2018.There were 348 males and 11 females(31.6:1 male and female)in this group.The average age of onset was 16.7±2.2 years,the average age of visits was 19.2±4.5 years,and the average duration of treatment was 29.3±45.4 months.Descriptive study of the clinical manifestations,radiologic and neurophysiological findings of this group of patients,the Huashan clinical diagnostic criteria of Hirayama disease were established by including 100% of the clinical manifestations,imaging and neurophysiological findings.According to the following parameters,the clinical classification system of Hirayama disease was proposed.The parameters specifically included:the muscle atrophy involves the upper limbs,whether the quadriplegia was active or hyperactive,the Babinski sign positive and other pyramidal tract damage,whether it was accompanied by sensory dysfunction such as upper limb numbness,muscle atrophy location,and the progress of clinical symptoms or electrophysiological examination within 6 months.Thirty patients were randomly selected from the above 359 cases.Four orthopedic surgeons who were not involved in the classification system completed the clinical classification within the specified time.The Kappa value was used for the credibility evaluation.Results　The Huashan diagnostic criteria of Hirayama disease included clinical manifestations,imaging examinations and neurophysiological examinations.The main diagnostic indicators were:1)occult onset puberty,more common in men;2)localized muscle atrophy and weakness in the upper extremities;3)compared with the cervical neutral MRI,the MRI of cervical flexion showed that spinal cord was significantly shift forward and the anterior spinal cord was narrowed or disappeared.4)MRI T2 weighting of the cervical flexion showed cyst-wall separation behind the spinal cord;5)Neurophysiological examination showed that the affected muscles were neurogenic damage.6)The affected parts are limited to the middle and lower neck segment.At the same time,it was necessary to combine imaging and neurophysiological manifestations to distinguish cervical spondylosis with upper limb muscle atrophy and motor neuron disease.According to the clinical characteristics of different patients,Hirayama disease can be divided into type I-III.Type I:72.1%,one-sided upper limb or one upper limb-based hand inner muscle and forearm muscle atrophy.According to whether progress of symptoms or electrophysiological examination was seen in the past 6 months,type I can be divided into:Ia.stable period.Regular follow-up assessment was recommended.If the disease progressed,to wear a cervical collar was suggested;surgery could be done if necessary;Ib.progression period,it was recommended to use a cervical collar,and regularly evaluate,if patients could not wear cervical collar for long,it was recommended to operate.Type II:14.2%,unilateral upper limb or one upper limb-based hand inner muscle and forearm muscle atrophy with pyramidal tract injury.Surgery was recommended.Type II:13.7%,atypical Hirayama disease,including upper limb proximal muscle atrophy,symmetrical double upper limb muscle atrophy,and sensory disturbances associated with upper limb numbness.Wear a cervical collar,and follow-up and assess closely,and choose surgical treatment if necessary.The credibility evaluation showed that the average Kappa value of the classification was 0.732(0.688-0.834),which is a basic credibility.Conclusion　The Huashan diagnostic criteria of Hirayama disease was conducive to the early diagnosis.The clinical classification system of Hirayama disease has good credibility and good clinical intervention guidance value.

Objective　To investigate the impact of the loss of attachment on the outcomes in Hirayama disease(HD)patients treated with anterior cervical discectomy and fusion(ACDF).Methods　A total of 26 patients(23 males and 3 females)who were diagnosed as HD and received 2 levels of ACDF surgery from February 2014 to November 2016 were enrolled in the present study.All patients took the MRI at both flexion and neutral position pre-operation and MRI at flexion position post-operation.The measured parameters related to performance of "loss of attachment" include:the distances between the posterior edge of the spinal cord and the cervical spinal canal(x),the anterior and posterior wall of the cervical spinal canal(y),the anterior-posterior(a)and the transverse diameter(b)of spinal cord cross sections.The value of the cervical spinal cord deformation(flexion position a/b)and the cervical spinal cord forward movement(flexion position x/y-neutral position x/y)were calculated,and the centrums refer to the performance of "loss of attachment" were also recorded.DASH scores and grip strength on both sides were recorded before and 1 year after operation.The 26 patients were divided into two groups,17 patients(17/26,65.4%)in improvement group with the decreased DASH scores and the other 9 patients(9/26,34.6%)in no improvement group.Logistic regression and ROC curve were used to analyze the influence factor of anterior cervical discectomy and fusion(ACDF)surgery on patients with Hirayama disease.Results　After surgery,the value of a/b significantly increased(t=4.27,P=0.001)and x/y significantly decreased(t=10.25,P=0.001).The performance of "loss of attachment" in 88.5%(23 of 26)patients disappeared after surgery,while the DASH score(P=0.06),and the grip strength of severe and mild side(P=0.36 and P=0.42)shew no obvious change.65.4%(17 of 26)patients shew a decrease in DASH scores after operation,the remaining 34.6%(9 of 26)patients shew no obvious change,several even with a mild increase.The segments refer to performance of "loss of attachment",the value of the cervical spinal cord de-formation and the cervical spinal cord forward movement shew a difference between the two group(t=-5.56,P=0.001;t=3.06,P=0.005;t=-3.76,P=0.001).The logistics regression analysis with the above three independent variable confirmed that the value of segments that refer to "loss of attachment" was the factor of the post-operative efficacy of ACDF surgery on patients with Hirayama disease(OR=6.963,P=0.001).And the ROC curve shew AUC=0.902,Jordan index=0.83,critical value=4.5.Conclusion　The anterior cervical discectomy and fusion(ACDF)surgery can effectively improve the loss of attachment at the surgical segments,and prevent the progress of upper limb dyskinesia.And range of the longitudinal separation refers to "loss of attachment" can impact the outcomes of ACDF surgery,more segments for fusion may be need in order to improve the outcomes of those patients with the range more than 5 segments.

Objective　To investigate the predictive value of dynamic F wave in the treatment of Hirayama disease(HD)after anterior cervical fusion and internal fixation.Methods　From February 2014 to January 2016,thirty-six patients with HD were included in this study(age:17.9±2.6 years,height:173.1±6.2 cm,disease duration:20.1±13.1 months;male to female ratio:35:1),and all of these 36 patients underwent anterior cervical autogenous iliac fusion and internal fixation.All of these patients underwent bilateral dynamic F-wave detection(The F-wave was tested on the cervical standard position and at least 30 minutes after cervical flexion)on the median nerve before operation and 3-5 days after operation.The latency and maximum amplitude of M-wave,the persistence of F-wave,the shortest latency of F-wave,the ratio of F/M and the persistence of repeated F-wave were measured.Dynamic F-wave abnormalities were defined when repeated F-wave was only found during neck flexion or the standard-flexion difference of each measurements of F-wave was more than 2 standard deviations from the normal value(the shortest latency of F-wave:0.6±0.5 ms,persistence of F-wave:7.3%±5.7%,the ratio of F/M:1.1±1.0).Furthermore,these patients with HD accepted both bilateral grip strength and hand function score(disabilities of arm,shoulder and hand,DASH)before and 1 year after operation.According to the presence of abnormal dynamic F-wave before operation,patients were divided into abnormal dynamic F-wave group and normal dynamic F-wave group.Fisher exact test was used to compare the percentage of the patient with improved DASH score in both patient groups after operation.Both DASH score and bilateral grip strength were analyzed by paired sample t test before and after operation.Results　Before operation,eleven(11/36,30.6%)patients with HD had abnormal dynamic F-wave,and there was no significant difference of age(t=-0.849,P=0.412),duration(t=1.110,P=0.282),DASH scores(t=2.002,P=0.055)and handgrips(more-symptomatic side:t=-0.673,P=0.507;symptomatic side:t=-1.729,P=0.094)between the patients with or without preoperative dynamic F-wave abnormalities.One-year follow-up demonstrated that significantly greater number of the patients presented with reduced DASH scores in the patients with preoperative dynamic F-wave abnormalities(10/11,90.9% vs 12/25,48.0%;P=0.025).Compared with the patients without preoperative dynamic F-wave abnormalities,the patients with preoperative dynamic F-wave abnormalities showed reduced DASH scores after operation(t=2.347,P=0.041).Furthermore,abnormal dynamic F-waves disappeared soon after operation in ten patients(90.9%,10/11)with preoperative dynamic F-wave abnormalities,and DASH scores reduced significantly in these ten patients after operation(4.4±3.8 vs 1.5±0.4,t=3.094,P=0.013),and the DASH score in another patient whose dynamic F wave did not disappear after operation increased significantly one year after operation(preoperative vs.postoperative:6.66 vs 9.87).Conclusion　Dynamic F-wave can be used as a preoperative evaluation method for HD to predict the recovery of upper limb function after operation.At the same time,the comparison of dynamic F-wave before and after operation is an effective and immediate method to evaluate the effectiveness of surgical treatment of HD.