This study constructed a LHCGR-CRE-luc-HEK293 transgenic cell line according to the activation of the cAMP signaling pathway after recombinant human chorionic gonadotropin binding to the receptor. The biological activity of recombinant human chorionic gonadotropin was assayed using a luciferase assay system. The relative potency of the samples was calculated using four-parameter model. And the method conditions were optimized to validate the specificity, relative accuracy, precision and linearity of the method. The results showed that there was a quantitative potency relationship of human chorinonic gonadotropin (hCG) in the method and it was in accordance with the four-parameter curve. After optimization, the conditions were determined as hCG dilution concentration of 2.5 μg·mL^-1, dilution ratio of 1∶4, cell number of 10 000-15 000 cells/well, and induction time of 6 h. The method had good specificity, relative accuracy with relative bias ranging from -8.9% to 3.4%, linear regression equation correlation coefficient of 0.996, intermediate precision geometric coefficient of variation ranging from 3.3% to 15.0%, and linearity range of 50% to 200%. This study successfully established and validated a reporter gene method to detect hCG biological activity, which can be used for hCG biological activity assay and quality control.

In this study, the CHO_INSR_1284 transgenic cell line was employed as the target cell, utilizing homogeneous time-resolved fluorescence technology to establish a method for detecting the biological activity of insulin degludec. Key parameters were optimized, and validation was conducted in accordance with general principles 9401 and 1431 of the fourth section of the 2020 edition of the Chinese Pharmacopoeia. Results indicated a good dose-response relationship for insulin degludec in this method, aligning with a four-parameter curve. Following optimization, the cell seeding density was set at 3.5×10⁵ cell·mL^-1, the initial concentration of insulin degludec at 57.18 μg·mL^-1, with a four-fold dilution, a stimulation period of 45 minutes, and an incubation duration of 4 hours. This method demonstrated strong specificity, with the geometric variation coefficient (GCV%) for the five potency levels ranging from 4.1% to 10.6%. The linear regression equation from the linear fitting was y = 1.015x - 0.027 7, and R² = 0.999 6. The results confirmed the method's good intermediate precision and linearity. The regression term was highly significant (P < 0.01), neither the deviation from parallel terms nor the model mismatch terms were significant (P ≥ 0.05), complying with general rule 1431 of the fourth section of the 2020 edition of the Chinese Pharmacopoeia. This study established a method for detecting the biological activity of insulin degludec using homogeneous time-resolved fluorescence technology, suitable for evaluating the biological activity and quality control of insulin degludec products.

Air Force Medical University has constructed an integrated clinical diagnosis curriculum system for undergraduate students based on military medical service and established the teaching methods of diagnostic evidence orientation, diagnostic method guidance, and diagnosed case practice. In the context of the new epidemic situation, we have also formed the blended teaching model of "triad training" and "four methods", optimized the teaching process of theory and practice, incorporated the contents of military medicine, determined the teaching objectives of this curriculum system at the three levels of knowledge, ability, and quality, completed the construction of the core curriculum, and applied it to the teaching practice. This curriculum system has achieved significant improvements in the degree of satisfaction among teachers and students and the excellent rate of military medical skill examination, which better meets the practical needs of medical service personnel training in air force.

On February 2022, WHO released the evidence-based guideline on control and elimination of human schistosomiasis, with aims to guide the elimination of schistosomiasis as a public health problem in disease-endemic countries by 2030 and promote the interruption of schistosomiasis transmission across the world. Based on the One Health concept, six evidence-based recommendations were proposed in this guideline. This article aims to analyze the feasibility of key aspects of this guideline in Chinese national schistosomiasis control program and illustrate the significance to guide the future actions for Chinese national schistosomiasis control program. Currently, the One Health concept has been embodied in the Chinese national schistosomiasis control program. Based on this new WHO guideline, the following recommendations are proposed for the national schistosomiasis control program of China: (1) improving the systematic framework building, facilitating the agreement of the cross-sectoral consensus, and building a high-level leadership group; (2) optimizing the current human and livestock treatments in the national schistosomiasis control program of China; (3) developing highly sensitive and specific diagnostics and the framework for verifying elimination of schistosomiasis; (4) accelerating the progress towards elimination of schistosomiasis and other parasitic diseases through integrating the national control programs for other parasitic diseases.
China/epidemiology* ; Disease Eradication ; Humans ; Public Health ; Schistosomiasis/prevention & control* ; World Health Organization

Objective: To explore the effect of moxibustion at Shenque (CV 8) on myocardial structure and function in exercise-induced fatigue rats. Methods: A 12-week treadmill running training was performed to create an exercise-induced fatigue rat model. Sixty eligible male specific-pathogen-free grade Sprague-Dawley rats were randomly divided into a blank group, a control group, a model group, a non-meridian non-acupoint group, a Zusanli (ST 36) group and a Shenque (CV 8) group, with 10 rats in each group. Rats in the blank group did not receive treadmill running training or moxibustion. Rats in the control group did not receive treadmill running training but received mild moxibustion at Shenque (CV 8). Rats in the model group received treadmill running training but no moxibustion. Rats in the non-meridian non-acupoint group, the Zusanli (ST 36) group and the Shenque (CV 8) group received moxibustion at the non-meridian non-acupoint points, Zusanli (ST 36) or Shenque (CV 8) immediately after each treadmill running training, 15 min each time, once a day for 5 consecutive days a week at a 2-day interval, 60 times of moxibustion in total. Left ventricular end-diastolic diameter (LVEDd), left ventricular end-systolic diameter (LVESd), left ventricular diastolic volume (LVDv), left ventricular systolic volume (LVSv), ejection fraction (EF), stroke volume (SV), early diastolic peak flow velocity of mitral valve (E) and late diastolic peak flow velocity of mitral valve (A) of each group before and after the last treadmill running training were measured. Blood was collected 6 h after the last treadmill running training, and serum C-reactive protein (CRP), myoglobin (Mb), creatine kinase-myocardial band (CK-MB), cardiac troponin I (cTnI) and cardiac troponin T (cTnT) levels were detected. Finally, the heart was separated, the heart mass (HM) was measured, the cTnT level of the myocardial tissue was detected, the ultrastructural changes of the left ventricular myocardium were observed by transmission electron microscope, the left ventricular fraction shortening (LVFS), E/A and heart mass index (HMI) were calculated. Results: Compared with the same group before treatment, the rat cardiac LVEDd, LVESd, LVDv, LVSv, SV, E and A were significantly increased (all P<0.01), and the rat LVFS, E/A and EF were significantly decreased (all P<0.01) in the model group and the non-meridian non-acupoint group after treatment; the rat cardiac SV, LVDv, LVSv, E and A were all increased (all P<0.01), while E/A and EF were decreased (all P<0.01) in the Zusanli (ST 36) group after treatment; the rat cardiac LVDv, E and A were significantly increased (P<0.01 or P<0.05), and E/A was significantly decreased (P<0.01) in the Shenque (CV 8) group after treatment. After treatment, compared with the blank group, the rat cardiac LVEDd, LVESd, SV, LVDv, LVSv, E, A, the serum CRP, Mb, CK-MB, cTnI, cTnT and HMI, and the myocardial cTnT were increased (all P<0.01), and the LVFS, E/A and EF were all reduced (all P<0.01) in the model group; compared with the model group and the non-meridian non-acupoint group, rats in the Zusanli (ST 36) group and the Shenque (CV 8) group showed decreased LVEDd, LVESd, SV, LVDv, LVSv, E, A, serum CRP, Mb, CK-MB, cTnI, cTnT and HMI, and myocardial cTnT (P<0.01 or P<0.05), along with increased LVFS, E/A and EF (all P<0.01); compared with the Zusanli (ST 36) group, Mb and A of the Shenque (CV 8) group were decreased (both P<0.01), while both E/A and EF were increased (P<0.01, P<0.05). Transmission electron microscopy examination showed that myofibrils in the blank group and the control group were neatly arranged with clear light and dark bands; the model group and the non-meridian non-acupoint group showed different degrees of myofibril disintegration and breakage, increased and aggregated mitochondria of different sizes, and increased electron density. The myofibrils in the Shenque (CV 8) group and Zusanli (ST 36) group were arranged neatly with clear light and dark bands, and compensatory hyperplasia of mitochondria. Conclusion: Moxibustion at Shenque (CV 8) and Zusanli (ST 36) both can effectively improve the occurrence of myocardial remodeling in exercise-induced fatigue rats, and the effect of moxibustion at Shenque (CV 8) is better in improving cardiac function.

OBJECTIVES: Since the outbreak of coronavirus disease 2019 (COVID-19), it has spread rapidly in China and many other countries. The rapid increase in the number of cases has caused widespread panic among people and has become the main public health problem in the world. Severe patients often have difficult breathing and/or hypoxemia after 1 week of onset. A few critically ill patients may not only rapidly develop into acute respiratory distress syndrome, but also may cause coagulopathy, as well as multiple organs failure (such as heart, liver and kidney) or even death. This article is to analyze the predictive role of clinical features in patients with COVID-19 for severe disease, so as to help doctor monitor the severity-related features, restrain the disease progress, and provide a reference for improvement of medical treatment. METHODS: The clinical data of 208 patients with COVID-19 who were isolated and treated in Changsha Public Health Treatment Center from January 17, 2020 to March 14, 2020 were collected. All patients were the mild and ordinary adult patients on admission, including 105 males and 103 females from 19 to 84 (median age 44) years old. According to the "Program for the diagnosis and treatment of novel coronavirus (COVID-19) infected pneumonia (Trial version 7)" issued by the General Office of National Health Committee and Office of State Administration of Traditional Chinese Medicine as the diagnostic and typing criteria. According to progression from mild to severe disease during hospitalization, the patients were divided into a mild group (=183) and a severe transformation group (=25). The clinical features such as age, underlying disease, blood routine, coagulation function, blood biochemistry, oxygenation index, and so on were analyzed. Among them, laboratory tests included white blood cell (WBC), lymphocytes (LYM), neutrophil (NEU), hemoglobin (Hb), platelet (PLT), prothrombin time (PT), plasma fibrinogen (Fib), activated partial prothrombin time (APTT), thrombin time (TT), -dimer, total bilirubin (TBIL), albumin (ALB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), serum creatinine (Cr), creatine kinase (CK), creatine kinase isoenzyme-MB (CK-MB), lactate dehydrogenase (LDH), C-reactive protein (CRP), and oxygen partial pressure in arterial blood. Partial pressure of oxygen in arterial blood/fractional concentration of inspiratory oxygen (PaO/FiO) was calculated. The variables with statistical significance were analyzed by logistic regression analysis. RESULTS: Patients in the severe transformation group had more combined underlying diseases than those in the mild group (<0.05). From the perspective of disease distribution, patients in the severe transformation group had more combined hypertension (<0.05). In the severe transformation group, PT was significantly longer, the levels of Fib, ALT, AST, CK, LDH, and CRP were significantly higher than those in the mild group (<0.05 or <0.001), while LYM, ALB, and PaO/FiO were significantly lower than those in the mild group (<0.05 or <0.001). Logistic regression analysis was performed on clinical features with statistically significant differences. Combined with hypertension, LYM, PT, Fib, ALB, ALT, AST, CK, LDH, and CRP as independent variables, and having severe disease or not was the dependent variable. The results show that combined hypertension, decreased LYM, longer PT, and increased CK level were independent risk factors that affected the severity of COVID-19 (<0.05). CONCLUSIONS The patients with mild COVID-19 who are apt to develop severe diseases may be related to combined hypertension, decreased LYM, and longer PT, and increased CK level. For the mild patients with these clinical features, early intervention may effectively prevent the progression to severe diseases.
Adult ; Aged ; Aged, 80 and over ; Betacoronavirus ; China ; Coronavirus Infections ; diagnosis ; physiopathology ; Disease Progression ; Female ; Hospitalization ; Humans ; Male ; Middle Aged ; Pandemics ; Pneumonia, Viral ; diagnosis ; physiopathology ; Retrospective Studies ; Young Adult

PURPOSE: To compare the efficacy and safety of recombinant human bone morphogenetic protein (rhBMP) and iliac crest autograft in the fusion treatment of lumbar spondylolisthesis. METHODS: The studies using randomized controlled trials to compare the rhBMP with iliac crest autograft in the treatment of lumbar spondylolisthesis were retrieved from Embase, Pubmed, ProQuest dissertations & theses (PQDT), China national knowledge infrastructure (CNKI), Chinese Biomedical Database, Wanfang Data, Cochrane Library (from March 1998 to March 2018). Postoperative fusion rate, clinical success rate, postoperative intervertebral height, complications, operation time, blood loss and duration of hospitalization were chosen as the outcome indicators. Methodological quality of the trials was critically assessed, and relevant data were extracted. Statistical software Revman 5.3 was used for data-analysis. RESULTS: Eleven articles were included in the meta-analysis. The results showed that, comparing the efficacy of rhBMP with iliac crest autograft, statistical significance was found in the 24-month fusion rate post operation [95% CI (1.38, 24.70), p = 0.02] and operation time [95% CI (-14.22, -2.08), p = 0.008]. There is not sufficient evidence for statistical differences in the remaining indicators. CONCLUSION The current literature shows rhBMP is a safe and effective grafting material in the treatment of lumbar spondylolisthesis. Further evidence is dependent on the emergence of more randomized controlled trials with higher quality and larger sample sizes in the future.
Autografts ; Bone Morphogenetic Proteins ; administration & dosage ; Databases, Bibliographic ; Humans ; Ilium ; transplantation ; Lumbar Vertebrae ; surgery ; Randomized Controlled Trials as Topic ; Recombinant Proteins ; administration & dosage ; Spinal Fusion ; methods ; Spondylolisthesis ; surgery ; Time Factors ; Treatment Outcome

Objective: To investigate the mRNA and protein expressions of outer dense fiber 2 (ODF2) in the sperm of the asthenospermia patient and their differences from those in normal healthy men. METHODS: According to the WHO criteria, we collected semen samples from 45 asthenozoospermia patients and 15 normal healthy volunteers. Using computer-assisted sperm analysis (CASA), we divided the semen samples from the asthenospermia patients into a mild, a moderate and a severe group, and determined the mRNA and protein expressions of ODF2 in different groups by RT-PCR and Western blot. RESULTS: Compared with the normal healthy men, the expression of the ODF2 gene showed no statistically significant difference in the mild asthenospermia group (1.112 0 ± 0.525 5 vs 0.688 0 ± 0.372 0, P >0.05) but remarkably decreased in the moderate (0.483 3 ± 0.186 3, P <0.05) and severe asthenospermia patients (0.448 3 ± 0.340 8, P <0.01). The OD value (ODF2/β-actin) of the ODF2 protein in the normal men exhibited no statistically significant difference from that in the mild asthenospermia group (0.458 7 ± 0.052 1 vs 0.326 1 ± 0.071 4, P >0.05), but markedly lower than in the moderate (0.145 4 ± 0.053 6, P <0.05) and severe asthenospermia patients (0.122 7 ± 0.045 7, P <0.01), which was consistent with the results of RT-PCR. CONCLUSIONS Decreased mRNA and protein expressions of ODF2 in the sperm are positively correlated with declined sperm motility of the asthenospermia patient, which is suggestive of the involvement of the ODF2 gene in the regulation of sperm motility.
Asthenozoospermia ; metabolism ; physiopathology ; Case-Control Studies ; Down-Regulation ; Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Male ; RNA, Messenger ; metabolism ; Semen Analysis ; Sperm Motility ; Sperm Tail ; Spermatozoa ; metabolism

OBJECTIVETo explore the mechanism of miR-124 inhibiting the proliferative activity of prostate cancer PC3 cells.

METHODSLuciferase reporter gene assay was used to examine the specific binding ability of miR-124 to PKM2 mRNA 3'-UTR. After miR-124 was transfected mimic to PC3 cells, the expression levels of PKM2 mRNA and protein were detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, respectively. The effects of miR-124 mimic and PKM2 siRNA on the proliferative activity of the PC3 cells were determined by MTT assay.

RESULTSThe expressions of PKM2 mRNA and protein were upregulated (5.12 +/- 0.35) times and (4.05 +/- 0.20) times respectively in the PC3 cells as compared with those in the RWPE-1 cells (P < 0.05). Luciferase reporter gene assay demonstrated that miR-124 targeted PKM2 3'-UTR. At 24 hours after transfection with miR-124 mimic, the PKM2 protein expression in the PC3 cells was downregulated (0.16 +/- 0.04) times (P < 0.05), while the PKM2 mRNA level was not changed significantly (P > 0.05), as compared with the control group. MTT assay showed that both miRNA-124 mimic and PKM2 siRNA could inhibit the proliferation of the PC3 cells, but the former exhibited a greater inhibitory effect than the latter. After transfection with miR-124 mimic and PKM2 siRNA, the cell growth rates were (66.20 +/- 5.10)% vs (82.10 +/- 6.35)% at 24 hours (P < 0.05) and (49.34 +/- 2.37)% vs (70.10 +/- 5.80)% at 48 hours (P < 0.05).

CONCLUSIONmiR-124 can suppress the proliferation of PC3 cells by regulating the PKM2 gene.

Carrier Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; genetics ; Humans ; Male ; Membrane Proteins ; genetics ; metabolism ; MicroRNAs ; genetics ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; Thyroid Hormones ; genetics ; metabolism ; Transfection

OBJECTIVETo explore the effects of cyclopamine on the proliferation and apoptosis of LNCaP cells and the expression of the PCA3 gene in human prostate cancer in vitro.

METHODSLNCaP cells were treated with cyclopamine at the concentrations of 1, 5, 10 and 15 micromol/L for 24, 48 and 72 hours. The inhibitory effects of cyclopamine on the proliferation and apoptosis of the LNCaP cells were detected by MTT and flow cytometry respectively, the morphological changes of the cells observed by Hoechst 33258 staining, and the expression of the PCA3 gene determined by real-time fluorescence quantitative reverse transcriptase polymerase chain reaction (FQ-RT-PCR).

RESULTSCompared with the blank control group, cyclopamine significantly inhibited the proliferation of the LNCaP cells at 5, 10 and 15 micromol/L (P <0.01), reaching IC50 at 10 micro mol/L at 48 hours. The apoptosis rates of the LNCaP cells at 24, 48 and 72 hours were 37.21%, 57.38% and 57.98% in the 10 micromol/L group and 21. 16% , 71.31% and 72.90% in the 15 micro.mol/L group, significantly different from those in the control (P <0. 01). The cell apoptosis showed a rising trend with the increase of cyclopamine concentration and acting-time, while the expression of the PCA3 gene was decreasing with the increased concentration of cyclopamine, significantly lower than that of the blank control group (P <0.01) , and extremely low in the 10 micromo/L group

CONCLUSIONCyclopamine intervention at 10 and 15 micromol/L for 48 and 72 hours could significantly inhibit the at all time points. Proliferation and induce the apoptosis of LNCaP cells and reduce the expression level of PCA3.

Antigens, Neoplasm ; genetics ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Male ; Prostatic Neoplasms ; genetics ; pathology ; Veratrum Alkaloids ; pharmacology