Objective To investigate the feasibility of a new polyclonal antibody specific to fetal hemoglobin (HbF) and its application in enrichment of circulating fetal nucleated red blood cell(NRBC) for non-invasive prenatal diagnosis. Methods A polyclonal antibody against a synthetic peptide comprising residues 69-78 of the γ-chain of HbF was prepared and conjugated to carrier protein KLH as the immunogen according to the specific antigenic determinant. The peptide-KLH solution was mixed with freund's complete or incomplete adjuvant and immunized goat to prepare specific polyclonal antibody against the γ-chain of fetal hemoglobin. After purification with protein G, maternal blood was obtained from 32 pregnant women at 22 to 39 weeks of gestation. NRBCs were separated and then stained with antibody against the γ chain of HbF. All the positive cells were collected by micromanipulator under microscopic observation, and whole genome was amplified by improved primer extension preamplification (PEP). Multiplex polymerase chain reaction amplification at nine different polymorphic short tandem repeat (STR) loci was also used to determine origin of the positive cells isolated from maternal blood. Results NRBCs stained with antibody against the γ chain of HbF were found in all of the blood from the 32 cases. Attached positive cells with anti-HbF staining have unique morphological characteristics, low nucleus-to-cytoplasm ratio, brown cytoplasm and blue dense nucleus after hematoxylin counterstain under microscopic observation, which can distinguished NRBCs with other cells. A total of 183 NRBCs were found in all of 32 pregnant women at a range of 0.6～1.8 cell/ml venous blood. The accurate rate was 90.6% by the STR genotype identification. Conclusion The antibodies specific to fetal γ-chain of fetal hemoglobin with synthetic peptide technology may have wide clinical utility in identification of fetal NRBCs from maternal circulation for non-invasive prenatal genetic diagnosis.

Objective:To investigate the changes of left ventricular mass index(LVMI),plasma brain natriuretic peptide (BNP),interJeukin-6(IL-6)and interleukin-10(IL-10)in patients with congestive heart failure(CHF),and their values in risk stratification and prognostic evaluation of CHF thereof.Methods:Ninety-five patients with CHF in accordance with 2001ACC/AHA guidelines were enrolled and stratified as stage A(n=21),B(n=23),C(n=31)and D(n=20).The levels of plasma BNP,IL-6 and IL-10 were measured and compaired with those in 20 healthy subjects(control group).The patients were followed up every three months in two years.The end point was the rehospitalization and the cardiac death.Results:(1)The level of LVMI increased early in patients with CHF at stage B.(2)The plasma BNP levels were initially elevated in CHF patients at stage B,and the concentrations of BNP closely paralleled disease severity.(3)The levels of plasma IL-6 and IL-10 were increased,and the ratio of IL-10/IL-6 was decreased,with the progression of CHF at stage C and D.The plasma BNP level was positively correlated with the levels of IL-6 and IL-10.(4)The new CHF stratification was surperior to NYHA functional classification in regard to CHF prognosis prediction.(5)Multivariate COX regression analysis showed the levels of BNP and IL-6 were the independent risk factors in prognostic evaluation(P＜0.01,P=0.026).(6)Patients in high-risk groups stratified by the combination of BNP and stage classification had obviously higher end point events rate.Conclusion:(1)LVMI was a better marker for the evaluation of left ventrieular remodeling.(2)The intrinsic mechanism of CHF progression was inflammatory reaction deterioration.(3)BNP was a good marker in predicting onset and prognosis of CHF.The combination of BNP and the stage classification could improve the prognostic evaluation in patients with CHF.

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the world, and the study on the regulatory mechanisms of the invasion and migration of HCC is of great significance to clinical diagnosis and treatment. Circular RNA (circRNA), as an important member of the non-coding RNA family, plays the role of microRNA (miRNA) sponge in hepatocytes due to its highly stable circular structure. It also plays an important role in HCC progression by regulating miRNA or promoting the expression of target genes through the competitive endogenous RNA mechanism. This article explores the mechanism of action of circRNA in the pathogenesis of HCC, so as to help with the screening for diagnostic markers of HCC and the development of effective therapeutic targets for HCC.

AIM: To explore the expressive role of lipoprotein-associated phospholipase A_2, high sensitive C-reactive protein and matrix metalloproteinase-9 in vulnerable atherosclerotic plaques in a rabbit model. METHODS: Forty eight New Zealand white male rabbits were randomly divided into 4 groups (12 rabbits each): control group, stable plaque group, p53 group, and p53+drug group. Rabbits in control group were fed with a regular diet and underwent sham operation. Rabbits in stable plaque group, p53 group and p53+drug group underwent balloon induced arterial wall injury and then were fed on a diet with 1% cholesterol. The animals were all fed for 3 months, then the rabbits in p53 group and p53+drug group underwent Ad5-CMV p53 transfection at 10th week. Before killed, the animals in p53+drug group underwent pharmacological triggering with Russell's viper venom (RVV) and histamine to induce the rupture of the atherosclerotic plaques. At the 1st day and before sacrifice, the serum was collected for measuring Lp-PLA_2, hs-CRP, MMP-9, HDL, LDL and VLDL. The expressions of Lp-PLA_2, hs-CRP and MMP-9 in tissues were determined by the methods of hybridization and immunohistochemistry. RESULTS: At the end of 12th week, the serum and tissue levels of Lp-PLA_2 and MMP-9 in stable plaque group, p53 group and p53+drug group were significant different from those in control group and in each group at the first day (P<0.05). The serum levels of Lp-PLA_2 and hs-CRP in p53 group and p53+drug group were significantly higher than those in control group and stable group (P<0.05). The serum levels of Lp-PLA_2, hs-CRP and MMP-9 were all significantly different between p53 group and p53+drug group (P<0.05). At the end of 12th week, pathological results showed that 4 groups were normal artery, stable plaque, vulnerable plaque and rupture plaque, respectively. The fabric cap was thicker in plaque groups than that in normal group (P<0.05). The rupture and formation of thrombus were more significant in p53+drug group than those in p53 group. The serum level of Lp-PLA_2 had negative interrelated relationship with fabric cap in plaque groups (r=-0.710, P<0.01), and hs-CRP, MMP-9 had no interrelated relationships with fabric cap in plaque groups. CONCLUSION: Base on the successful establishment of the atherosclerotic plaque animal model, serum Lp-PLA_2 shows better interrelated relationships to plaques stability. Combination with hs-CRP and MMP-9, we can exactly evaluate the nature of plaques.

w biomarker to predict the presence of vulnerable plaque.

Objective:To investigate the diagnostic and prognostic value of the growth differentiation factor 15 (GDF15) and the procalcitonin (PCT) in sepsis.Methods:A total number of 137 patients with sepsis (considered as the sepsis group) and 59 patients with inflammatory infection but not diagnosed as sepsis (the non-sepsis group) received treatment in intensive care unit of Renming Hospital of Wuhan University were collected from July 2020 to January 2021, and 62 cases of healthy physical examination (control group) were simultaneously chosen as control. Sepsis patients were divided into two groups (death group [ n=48] and survival group [ n=89]) according to their 28-day′s survival. The serum levels of GDF15, PCT, C-reactive protein (CRP), interleukin-6 (IL-6) and interleukin-10 (IL-10) were examined, and the levels of each index, was dynamically monitored on the 1st, 3rd and 7th day after admission. The differences of the two indicators between different groups were compared by non-parametric test. The correlation between GDF15 and PCT was analyzed by Spearman correlation test. The receiver operating characteristic (ROC) curve was drawn to evaluate the diagnostic and prognostic value of the two indicators for sepsis. Results:The levels of GDF15 in the sepsis group, non-sepsis group and control group were 3.22 (1.39, 6.31) μg/L, 0.84 (0.21, 1.66) μg/L and 0.11 (0.09, 0.13) μg/L, respectively. The levels of PCT were 13.10 (1.99, 50.25) μg/L, 0.24 (0.13, 0.68) μg/L and 0.05 (0.03, 0.10) μg/L, respectively. The levels of CRP were 115.80 (26.40, 184.07) mg/L, 24.20 (11.30, 53.20) mg/L and 0.50 (0.50, 2.76) mg/L, respectively. The levels of IL-6 were 68.26 (21.59, 255.46) ng/L, 33.20 (10.81, 89.27) ng/L and 8.82 (7.33, 11.23) ng/L, respectively. The levels of IL-10 were 11.30 (5.88, 25.50) ng/L, 9.34 (5.65, 16.90) ng/L and 4.94 (4.31, 5.31) ng/L, respectively. The GDF15, PCT, CRP and IL-6 of the sepsis group were significantly higher than those of the non-sepsis group (The U values were 67.681, 86.034, 44.164 and 38.934, respectively, with P values less than 0.05) and the control group (The U values were 136.475, 138.667, 120.701 and 100.886, respectively, with P values less than 0.001). There was no significant difference in IL-10 between sepsis group and nonsepsis group, but it was higher than that of control group ( U=80.221, P<0.001). There was a positive correlation between GDF15 and PCT in patients with sepsis, and the spearman correlation coefficient was 0.234 ( P=0.006). The GDF15 of the death group and the survival group were 5.49 (3.60, 8.25) μg/L and 2.03 (1.06, 3.69) μg/L, and the PCT levels were 26.45 (11.23, 94.25) μg/L and 9.08 (1.33, 22.75) μg/L, respectively. GDF15 and PCT in the death group were significantly higher than those in the survival group ( U values were 3 305.500 and 3 060.000, respectively, and P values were both less than 0.001). The GDF15 and PCT levels in the death group were higher than those in the survival group on the 1st, 3rd and 7th day of dynamic monitoring ( P<0.05), however, the level of CRP and IL-10 were not significantly different ( P>0.05). The level of IL-6 in the death group was not significantly different from that of the death group on 1st day, but was higher than that of the survival group on the 3rd and 7th day ( P<0.05). The area under the curve (AUC) of GDF15, PCT, CRP, IL-6 and IL-10 alone and in the combined diagnosis of sepsis were 0.899, 0.938, 0.874, 0.789, 0.698 and 0.962, respectively. The combined detection of AUC was better than a single index; the GDF15, PCT, CRP, IL-6 and IL-10 alone and combined detection of sepsis prognosis AUC were 0.774, 0.716, 0.522, 0.623, 0.520 and 0.839, respectively, the combined detection of AUC is also better than single index. Conclusions:GDF15 and PCT have good clinical reference value in the differential diagnosis and prognosis of sepsis. The combination of indicators has a higher clinical value. GDF15 may become a biomarker for the diagnosis and prognosis of sepsis.

BACKGROUND:There are increasing scientific and technological ways to promote the recovery of muscle fatigue after sports,such as vibrating foam shaft,fascia gun relaxation,and ultra-low temperature cold therapy.Fascia gun relaxation has been widely used in practice,but there is still relatively little research on fascial guns. OBJECTIVE:To compare the effects of three relaxation methods(fascia gun,stretching,fascia gun+stretching)on muscle status and muscle strength during the recovery of exercise-induced muscle fatigue,and to provide scientific basis for the application of the three methods in practice. METHODS:Forty college students were randomly divided into control group(n=10),stretching group(n=10),fascia gun group(n=10),and fascia gun+stretching group(n=10).All subjects completed leg flexion and extension training at 60%1 RM,15 times per set,for 10 sets in total,to make exercise fatigue models.After modeling,the subjects were intervened with supine rest,static stretching,fascia gun relaxation,fascia gun+stretching relaxation respectively.Muscle status and muscle strength indexes were tested before exercise,immediately after exercise,immediately after relaxation,24 and 48 hours after exercise. RESULTS AND CONCLUSION:The muscle tension and dynamic hardness of the control group and the stretching group immediately after relaxation were significantly higher than those before training(P<0.01).However,there was no significant difference in the fascia gun group and fascia gun+stretching group before and after training(P>0.05).At 24 hours after training,the peak torque of the control group and fascia gun group was significantly lower than that before training(P<0.01),and there was no significant difference in the stretching group and fascia gun+stretching group before and 24 hours after training(P>0.05).To conclude,the fascia gun can immediately and effectively improve the muscle state of sports fatigue and stretching can promote the effective recovery of muscle strength within 24 hours after sports fatigue.Relaxation using fascia gun plus stretching can achieve the superposition of the two effects.

Objective To study the effect of maintenance of handpiece by manual filling and machine oil injection method , so as to perfect the manual filling process .Methods Totals of 600 extraction used handpiece after cleaned and dried were randomly divided into six groups .Group A was given manual filling , Group B1 ,B2 ,B3 ,B4 were given manual filling and pressure air gun to blow the handpiece cavity for 1-2,4-5, 9-10,14-15 seconds.Group C used oil filling machine .To evaluate the effect of maintenance of handpiece , the turbo-rotors bearing surface traces after sterilization with a microscope were observed , and the remaining oil injection weight was weighed and the oil area of handpiece packaging sterilization bags was measured . Results Turbo-rotors bearings surfaces were visible traces of lubricating oil left after sterilization for all the six groups.The comparison results of the remaining oil weight of turbo-rotors bearing surfaces showed that group C and group A, B1, B3, B4 were significantly different (P<0.05), but group C and group B2 had no significant difference (P>0.05).The comparison results of the oil area of handpiece packaging sterilization bags showed that group C and group A, B1, B3 tail, B4 had significant differences (P<0.05), while group C and group B2, B3 head had no significant difference (P>0.05).Conclusions The effect of maintenance of machine oil is better than pure manual filling .The perfecting manual filling process of maintenance of handpiece , which is manual filling , which using the pressure air gun to blow the handpiece cavity for 4-5 seconds , can achieve the equivalent maintenance effect with the machine oil injection .

Objective To investigate the clinical value of alpha-fetoprotein (AFP) combined with gamma-glutamyl transpeptidase (GGT)/aspartate aminotransferase (AST) ratio in the diagnosis of hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC). Methods A total of 352 subjects who received treatment or underwent physical examination in Renmin Hospital of Wuhan University from January 15 to June 15, 2020, were enrolled, among whom there were 86 healthy controls (HC group), 68 patients with chronic hepatitis B (CHB group), 69 patients with liver cirrhosis (LC group), and 129 patients with HCC (HCC group), and a retrospective analysis was performed for the serological test results of all subjects. An analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups; the Kruskal-Wallis H test was used for comparison of continuous data with skewed distribution between multiple groups, and the Nemenyi method was used for further comparison between two groups. A binary logistic regression analysis was used to calculate predictor variables; a receiver operating characteristic (ROC) curve was plotted for AFP, GGT/AST, and the predictor variables used alone or in combination, and the area under the ROC curve (AUC), sensitivity, and specificity were calculated; the Z test was used for comparison of AUC. Results The HCC group had significantly higher GGT/AST ratio and AFP than the other groups (all P < 0.05). The ROC curve analysis showed that AFP combined with GGT/AST ratio had a significantly higher AUC than AFP alone in the HCC group vs the LC group, the HCC group vs the HC+CHB+LC groups, and the HCC group vs the CHB+LC groups ( Z =2.684, 2.241, and 2.415, P =0.007, 0.025, and 0.016). Conclusion AFP combined with GGT/AST ratio can improve the clinical diagnostic performance of HBV-related HCC and thus has a certain diagnostic value.

Background: The Lymphadenectomy in Ovarian Neoplasms (LION) study revealed that systemic lymphadenectomy did not bring survival benefit for advanced ovarian cancer patients with clinically normal lymph nodes and was associated with a higher incidence of operative complications. However, there is no consensus on whether lymphadenectomy has survival benefit or not in early epithelial ovarian cancer (EOC). Methods We designed the LOVE study, a multicenter, randomized controlled, phase III trial to compare the efficacy and safety of comprehensive staging surgery with or without lymphadenectomy in stages IA-IIB EOC and fallopian tube carcinomas (FTC). The hypothesis is that the oncological outcomes provided by comprehensive staging surgery without lymphadenectomy are non-inferior to those of conventional completion staging surgery in early-stage EOC and FTC patients who have indications for post-operative adjuvant chemotherapy. Patients assigned to experimental group will undergo comprehensive staging surgery, but lymphadenectomy. Patients assigned to comparative group will undergo completion staging surgery including systematic pelvic and para-aortic lymphadenectomy. All subjects will receive 3–6 cycles of standard adjuvant chemotherapy. Major inclusion criteria are pathologic confirmed stage IA-IIB EOC or FTC, and patients have indications for adjuvant chemotherapy either confirmed by intraoperative fast frozen section or previous pathology after an incomplete staging surgery. Major exclusion criteria are non-epithelial tumors and low-grade serous carcinoma. Patients with severe rectum involvement which lead to partial rectum resection will be excluded. The sample size is 656 subjects. Primary endpoint is disease-free survival.