Objective To investigate the role of miR-194-3p in regulating functional changes in osteoblasts in a simulated microgravity environment and to provide a theoretical foundation for understanding the mechanical response mechanisms of osteoblasts in extreme mechanical environments.Methods The effects of microgravity on osteoblasts were simulated by using a rotary cell culture system.MC3T3-E1 osteoblasts were transfected with an miR-194-3p inhibitor and changes in proliferation,differentiation,apoptosis,and mineralization were assessed using MTT assay,RT-PCR,Western blot,double fluorescence staining,and alizarin red staining.Results Elevated expression of miR-194-3p under simulated microgravity conditions led to the suppression of osteoblast proliferation,differentiation,and mineralization to a certain extent,while promoting osteoblast apoptosis.However,transfection with the miR-194-3p inhibitor significantly downregulated miR-194-3p expression and partially reversed the reduced osteoblast proliferation,decreased expression of osteogenic differentiation markers such as ALP,OCN,and COL-I genes and proteins,decreased bone mineralization nodules,and increased osteoblast apoptosis induced by microgravity exposure.These findings indicated that miR-194-3p effectively ameliorates abnormal osteoblast function under microgravity conditions.Conclusions MiR-194-3p acts as a negative regulatory factor in the mechanical responses of osteoblasts under simulated microgravity.

Objective:To compare the changes of serum calcium level before and after surgical resection in patients with primary hyperparathyroidism.Methods:Two hundred and seventy-one patients with primary hyperparathyroidism were enrolled from Dec 1992 to Dec 2020 in Beijing Jishuitan Hospital. Serum calcium concentrations were measured before operation, 20 min during surgery, then 2 weeks 1-6 months , 7-12 months and 1 year respectively after operation. The baseline data of postoperative serum calcium such as sex, age, other genetic endocrine diseases, osteopathia and urolithiasis were calculated. The generalized estimation equation was used to analyze the changes of serum calcium in different types of patients before and after operation.Results:The most common postoperative hypocalcemia occurred within 2 weeks, and it occurred frequently half a year after surgery. There was no significant difference in blood calcium between male patients ( t=0.875, P=1.000) and patients with bone lesions ( t=0.034, P=3.049) from 1 to 6 months after surgery and 2 weeks after surgery. Blood calcium level in patients aged 15-35 years old from 1 to 6 months ( t=0.239, P=1.000) , from 7 to 12 months ( t=1.380, P=0.935) and 2 weeks after surgery was not statistically different. The change of bone mineral density was correlated with the change of blood calcium after operation ( F=6.895, P=0.004). Conclusions:The incidence of hypocalcemia was the highest in patients with hyperparathyroidism 2 weeks after surgery, and the blood calcium level was stable within the normal range 1 year later. The blood calcium value of male patients was still at a lower level than that of female patients within six months after surgery. In patients with bone disease, the blood calcium value was lower and recovered slowly 2 weeks after surgery. The blood calcium value of patients aged 15-35 was at a low level within 1 year after surgery.

Objectiv:To evaluate ultrasound, radionuclide imaging and CT in preoperative localization diagnosis of primary hyperparathyroidism (PHPT).Method:A total of 170 PHPT patients admitted to the hospital between Jan 1992 and Dec 2020 were analyzed retrospectively. The preoperative localization diagnostic efficacy of ultrasonography, radionuclide and CT alone and in combination was compared in groups.Results:The overall sensitivity of ultrasound, radionuclide and CT were 82.13%,80.43% and 75.74%. For normal positioned parathyroid adenoma: as for sensitivity of location diagnosis, ultrasound (86.67%) was higher than radionuclide (81.82%, P<0.05) and CT (80.59%, P<0.05), ultrasound/CT parallel test (94.70%, P<0.05) was higher than ultrasound alone. For specificity of location diagnosis, radionuclide (97.78%) was higher than ultrasound (91.62%) and CT (93.39%), both ultrasound/radionuclide series tests (99.00%, P<0.001)and ultrasound/CT series tests (96.94%, P<0.001) were higher than ultrasound alone. In case of ectopic parathyroid adenoma and parathyroid hyperplasia: the sensitivity and specificity of radionuclide seemed higher than ultrasound and CT. Conclusions:Ultrasound is the first choice for preoperative location diagnosis of PHPT. Ultrasound combined with radionuclide or CT can significantly improve the diagnostic efficiency of parathyroid lesions.

Objective:To summarize clinical characteristics of and treatment experience with patients with critical illnesses in a dermatological ward.Methods:All patients with serious or life-threatening conditions, who were hospitalized at the dermatological ward of the Second Xiangya Hospital of Central South University from July 9, 2011 to December 31, 2020, were collected, and their clinical data were retrospectively analyzed. Demographic characteristics, disease types and proportions, main complications, causes of serious or life-threatening conditions, important treatment measures and outcomes were summarized, and causes of death were also analyzed and discussed.Results:A total of 1 057 patients with critical illnesses were collected, with a male-to-female ratio of 1∶1.11, and 64.81% of them aged 18 to 65 years. The types of diseases mainly included drug eruptions (332 cases) , connective tissue diseases (226 cases) , bullous skin diseases (104 cases) , psoriasis (57 cases) , erythroderma (45 cases) , infectious skin diseases (67 cases) , etc. Among them, psoriasis (39 cases) and erythroderma (32 cases) mostly occurred in males, and connective tissue diseases (168 cases) mostly occurred in females. Common complications mainly involved infections, important organ damage or dysfunction, hypoalbuminemia, and fluid, electrolyte and acid-base imbalances. A total of 94 patients were diagnosed with life-threatening conditions, which were found to be mainly caused by primary skin diseases, hematologic abnormalities, respiratory failure, nervous system abnormalities, renal failure, sepsis, fluid, electrolyte and acid-base imbalances, etc. During the management of critical illnesses, 43 patients were treated with high-dose glucocorticoid pulse therapy, 264 were treated with gamma-globulin pulse therapy, 355 were transfused with other blood products, and 34 received special therapies such as hemoperfusion/immunoadsorption therapy, plasma exchange, dialysis, artificial liver support therapy; 42 patients were transferred to the intensive care unit (ICU) , 12 were transferred to the department of surgery for operations, and 12 were transferred to the department of obstetrics and gynecology for delivery or induction of labor. After treatment, 989 patients (93.57%) achieved improvement and were discharged. A total of 14 patients (1.32%) died, of whom 7 died of secondary sepsis, 2 died of severe pulmonary infections, 2 died of asphyxia caused by respiratory mucosa shedding-induced airway obstruction, the other 3 died of gastrointestinal hemorrhage, cerebral hemorrhage and neuropsychiatric systemic lupus erythematosus, respectively.Conclusions:Critical cases in the dermatological ward mainly suffered from serious skin diseases such as severe drug eruptions, connective tissue diseases and bullous skin diseases, as well as complications such as severe underlying diseases, severe organ dysfunction, sepsis or severe fluid, electrolyte and acid-base imbalances. In terms of treatment, it is of critical significance to make a clear diagnosis and assess the severity of disease as early as possible, monitor and prevent possible complications, and to consult with specialists in relevant disciplines in time.

Vaccination is the most effective way to prevent influenza and severe outcoming caused by influenza viruses.Health care workers(HCW) are exposed to patients with influenza and they are at high risk of occupationally acquired influenza and of causing nosocomial infection among patients,increasing the incidence rate,the risk of severe and death of patients.Improving the influenza immunization in HCW can not only reduce the prevalence of themselves and keep a weel-oiled of health care facilities during the influenza seasons,but also reduce the risk of severe and death among patients and increase the influenza vaccine uptake in whole population.At present,the influenza immunization coverage of HCW is low.The obstacles and myths of influenza vaccine are barriers for vaccine uptake among HCW.The various strategies are critical in order to improve the influenza coverage rates of HCW.

Objective To express UL148 RNA of human cytomegalovirus (HCMV) clinical strains in vitro and to study its functions. Methods Urine of a newborn with HCMV infection was inocula-ted into human embryo lung cells. HCMV clinical strain was isolated and identified by multiplex PCR. UL148 gene was amplified and cloned into pGEM-T-Easy plasmid after double enzyme digestion. A recombi-nant plasmid was constructed and located at the downstream of the T7 promoter. The recombinant plasmid was identified by electrophoresis of the recombinant plasmid,PCR product and double enzyme product. Se-quencing analysis was used for final confirmation. UL148 was transcribed into RNA by 32P labeling. Post-translational modification sites were analyzed by bioinformatics method based on UL148 sequence characteris-tics. Results The clinical strain of HCMV was obtained in vitro. Electrophoresis and sequencing analysis confirmed the successful construction of the recombinant plasmid. UL148 RNA was transcribed in vitro by T7RNA polymerase. Post-translational modification sites showed that UL148 gene contained one cell adhe-sion sequence, one legume lectins beta-chain signature, two N-myristoylation sites, one casein kinase Ⅱphosphorylation site,seven protein kinase C phosphorylation sitse, one cAMP/cGMP-dependent protein ki-nase phosphorylation site, two N-glycosylation sites and one transmembrane region. Conclusion UL148 gene might encode a viral adhesion molecule involving in the signal transduction in host cells.

Objective: To investigate the sensitivity and specificity of commercial nonstructural protein 1 (NS1) testing kits for Dengue fever diagnose, and provide the evidence for diagnostic criteria revision. Methods: 300 PCR or virus isolation positive blood samples for dengue virus were collected from sentinel hospitals for dengue surveillance in Guangzhou, Dongguang and Zhongshang from May 2015 to Nov. 2016. At the same time, 308 PCR negative samples for Dengue virus were collected as control group. The information of the sample was collected using questionnaires. These samples were tested using imported and domestic ELISA and the colloidal gold-labeled kits that were widely used for detecting dengue NS1. Sensitivity, specificity and coincidence were calculated and analyzed, and Z hongshan's result was regarded as the reslut of the third part. Results: The positive group includes 133 males and 167 females, average ages are 47.2±13.3, 179, 110 and 11 of them is Dengue Ⅰ, Ⅱ and Ⅲ respectively. The negative group includes 154 males and 154 females, average ages are (40.1±11.6) years old. The sensitivity of domestic ELISA Kits (94.5%) is less than imported (99.5%), and the result has statistical significance (χ²=8.59, P=0.030), the specificity is 99.7% and 97.7% respectively; The sensitivity of imported and domestic the colloidal gold-labeled Kits is 97.5% and 96.5% respectively, both of specificities are 100%. The sensitivity and specificity of Dengue Ⅰ for NS1 test are more than 97.0%. The sensitivity of domestic ELISA and gold-labeled Kits is 90.0% and 95.0%, and the specificity is 96.8% and 100% respectively for Dengue Ⅱ test. The sensitivity of imported ELISA and gold-labeled Kits is 100% and 98.0%, and the specificity is 99.4% and 100% respectively for Dengue Ⅱ test. The result of the third party show the sensitivity and specificity of domestic ELISA and gold-labeled Kits are 90.0% and 98.0%, the differences has statistical significance (χ²=5.67, P=0.020). Conclusion NS1 testing can be used as early dengue fever diagnose for higher sensitivity and specificity.

Objective To assess the clinical therapeutic effects and safety of Jiangtang Shuxin decoction (JTSXD) on diabetic patients complicated with chronic heart failure (CHF),and to search for its possible function mechanisms.Methods A prospective randomized controlled study was conducted,80 diabetic patients complicated with CHF [New York Heart Association (NYHA) functional class Ⅱ-Ⅲ] admitted into the Department of Traditional Chinese Medicine (TCM) or of Cardiology in Affiliated Hospital of Guangxi Youjiang National Medical College from October 2015 to September 2016 were enrolled,they were assigned to an observation group and a control group by randomized method with a computer,and finally 77 patients (39 cases in observation group and 38 cases in control group) completed this trial.The patients in control group received standardized routine western medical treatment,while the observation group was additionally administered JTSXD (including ingredients:astragalus 15 g,ginseng 10 g,radix ophiopogonis 15 g,radix rehmanniae 15 g,comus 10 g,rhizome coptidis 8 g,peach kernel 10 g,salvia mitiorrhiza 10 g,magnoliaceae 10 g,yam 15 g) on the basis of conventional therapy.The therapeutic course for all the patients in both groups was 2 months.Before and after treatment,the 6-minute walking distance (6MWD) was assessed;the TCM syndrome accumulated scores of the two groups were calculated;the left ventricular end-diastolic volume (LVEDV),the left ventricle ejection fraction (LVEF),the stroke volume (SV),the cardiac output (CO),and the maximum blood flow velocity of early diastolic/atrium late diastolic (E/A) were detected by echocardiography.The serum levels of glycosylated hemoglobin (HbA1c),angiotensin Ⅱ (Ang Ⅱ) and plasma B type brain natriuretic peptide (BNP) were tested with enzyme linked immunosorbent assay (ELISA);the level changes of total cholesterol (TC),triglyeride (TG),high density lipoprotein cholesteral (HDL-C) and low density lipoprotein cholesteral (LDL-C) were observed.Results Compared with the control group,after treatment in the observed group,the TCM syndrome score of palpitation,fatigue and thetotal accumulated score were all obviously decreased (palpitation score:0.9 ± 0.4 vs.1.2 ± 0.8,fatigue score:1.1 ± 0.7 vs.1.7 ± 0.8,total accumulated score:4.8 ± 1.2 vs.8.1 ± 1.8,all P ＜ 0.05);the LVEDV,the serum levels of HbA1c,Ang Ⅱ and BNP were also obviously decreased in the observed group [LVEDV (mL):136.28 ± 17.52 vs.158.82 ± 19.03,HbA1c (％):6.11±0.36 vs.6.89 ±0.32,Ang Ⅱ (ng/L):66.48 ± 17.64 vs.84.55 ± 20.39,BNP (μg/L):138.45 ± 87.55 vs.219.14±88.83,all P ＜ 0.05];The 6MWD,LVEF,SV,CO and E/A were all increased plainly in the observed group [6MWD (m):470.47 ± 79.66 vs.428.46 ± 88.56,LVEF:0.51 ±0.05 vs.0.46 ± 0.04,SV (mL):55.36 ± 2.88 vs.50.32±2.76,CO (L/min):5.74±0.91 vs.4.92±0.74,E/A:1.18±0.27 vs.0.83±0.28,all P ＜ 0.05].The degrees of decreased levels in TC,TG,LDL-C and the degrees of increased levels of HDL-C in observed group were superior to those of the control group,but there were no statistical significant differences (all P ＞ 0.05).Conclusion JTSXD shows good therapeutic effect and safety for treatment of diabetic patients accompanied by CHF (NYHA functional class Ⅱ-Ⅲ),and its mechanisms may be related to its regulation of glucose (reduction of HbA1c level),correction of lipid metabolism disorders,improvement of myocardial energy supply,inhibition of the activation of renin-angiotensin-aldosterone system (RAAS) and the secretion of BNP.

Objective: To develop a micro-neutralization test for determination of neutralizing antibody against ZIKA virus (ZIKV) in human sera and to verify the acute and convalescent serum samples of 10 ZIKA virus-infected cases diagnosed by nucleic acid detection and/or virus isolation. Methods: ZIKV isolated from ZIKA cases was used to determine micro-neutralization antibody. The virus solution was prepared by infecting BHK21, VERO and VERO-E6 cell lines and viral titer was tested; 100 TCID₅₀ viral solution and 4 times diluted sera which were inactivated at 56 ℃ for 30 min were neutralized, then added the cell suspension and incubated in 5% CO₂ incubator at 37 ℃ for 7 d. The CPE was observed every day. Results: The sensitivity of BHK21, VERO and VERO-E6 was different after infection with ZIKA virus. VERO cell line was the most sensitive and showed typical CPE. VERO cell line was used to establish a micro-neutralization test for determination of neutralizing antibody against ZIKA virus in sera. Conclusions The neutralizing antibody test for zika virus in sera is a special and usefulmethod to diagnose human infection of ZIKV and to conduct population based epidemiological investigation.

Objective To provide scientific evidences for Zika virus detection by clarifying the means by which Zika virus was discharged and the duration of corresponding processes. Methods Various samples of Zika cases were collected at different times and detected by using real-time RT-PCR. The positive samples were inoculated into cells and suckling mice through intracranial injection. The whole genome se-quences of those isolated Zika virus strain were sequenced and the results were further analyzed by comparing with the sequences of Zika virus from GenBank. Results The positive rates of Zika virus in urine, saliva and serum samples were 82. 4% (14/17), 82. 4% (14/17) and 52. 9% (9/17) respectively. The longest period of detected presence of Zika virus was found in urine samples amongst the three types of samples, fol-lowed by saliva and serum samples. Six Zika virus strains were isolated from 9 positive serum samples. Phy-logenetic analysis showed that the six genomes of Zika virus all belonged to Asia lineage, but located in two branches by Samoa and Venezuela strains. Conclusion This study indicated that urine, saliva and serum all could be used as the samples for routine detection of Zika virus. Urine and saliva samples showed higher detection rates of Zika virus RNA in comparison to serum samples, while Zika virus could be easily isolated from positive serum samples. Suckling mice were better for Zika virus isolation than cell lines.