Objective To obtain the heavy chain variable region (VH) gene of monoclonal antibody 2F2 against Japanese encephalitis virus (JEV).Methods Total RNA was isolated with Trizol from hybridoma 2F2 cells,and cDNA of VH was amplified with reverse transcription polymerase chain reaction (RT-PCR) and sequenced.The putative VH gene was expressed in E.coli,and the expressed products was detected with enzyme linked immunosorbent assay (ELISA) to determine the activity to bind JEV.Results The VH gene was 354 bp in length which encodes 118 amino acids.Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the VH gene was successfully expressed and purified from inclusion bodies.ELISA result also demonstrated that VH gene expressed products bind purified JEV.Conclusions The VH gene of monoclonal antibody 2F2 against JEV had been cloned.

Hydronephrosis is a common and frequent clinical disease. With the extensive application of minimally invasive techniques of Western medicine, it can quickly remove the obstruction and eliminate water. However, there may be problems of easy recurrence after surgery, causing more serious irreversible kidney damage. TCM has a unique advantage in the treatment of hydronephrosis, which can radically relieve the pain of patients, prevent recurrence, and protect the kidney function. Professor SHAO Zhao-di thinks that the cause of hydronephros is deficiency of kidney qi, which affects the function of gasification. Therefore, the treatment should be nourishing qi to invigorate spleen and reinforcing kidney for diuresis. In clinical practice, Bixie Fenqing Decoction can be used to treat hydronephrosis, which can obtain good efficacy.

Coronary heart disease is a kind of heart disease that is caused by atherosclerosis. The lipid deposition in the vessel wall results in occlusion of coronary artery and stenosis, which could induce myocardial ischemia and oxygen deficiency. Intervention therapies like percutaneous coronary intervention (PCI) and coronary stent improve myocardial perfusion using catheter angioplasty to reduce stenosis and occlusion of coronary artery lumen. Accordingly, intervention therapies are widely applied in clinic to treat ischemic cardiovascular disease, arterial intima hyperplasia and other heart diseases, which could save the patients' life rapidly and effectively. However, these interventions also damage the original endothelium, promote acute and subacute thrombosis and intimal hyperplasia, and thus induce in-stent restenosis (ISR) eventually. Studies indicated that the rapid reendothelialization of damaged section determined postoperative effects. In this review, reendothelialization of implants after intervention therapy is discussed, including the resource of cells contributed on injured artery, the influences of implanted stents on hemodynamic, and the effects of damaged degree on reendothelialization.
Angioplasty, Balloon, Coronary ; Cardiac Catheterization ; Coronary Artery Disease ; therapy ; Coronary Restenosis ; prevention & control ; Endothelium, Vascular ; pathology ; Humans ; Myocardial Ischemia ; prevention & control ; Stents ; Thrombosis ; prevention & control

Background Quartz dust cannot be degraded in the lungs, and inhalation of a large amount of quartz dust in the occupational production process will lead to the occurrence of pulmonary fibrosis, and then develop into silicosis. In recent years, studies have found that exosomes may be involved in the pathogenesis of fibrotic diseases by carrying microribonucleic acid (miRNA), but the mechanism of their actions in silicosis still needs to be studied. Objective To investigate the role of exosome-derived miRNA-21-5p/mothers against decapentaplegic homolog 7 (Smad7) in quartz dust-induced pulmonary fibrosis in rats. Methods Twenty-four healthy male SD rats were randomly divided into four groups (six rats in each group): control 4-week group, control 16-week group, quartz 4-week group, and quartz 16-week group. At the beginning of the experiment, 1 mL of quartz suspension (50 mg·mL⁻¹) and 1 mL of normal saline were injected into the trachea of rats in the quartz group and the control group, respectively, by means of one-time non-exposure intratracheal dust staining. Alveolar lavage was performed at the 4th and 16th weeks after dust staining, the exosomes in lavage solution were extracted by polyethylene glycol (PEG) precipitation, morphological identification was conducted by transmission electron microscopy (TEM), particle size of exosomes was detected by nano-tracking analysis (NTA), and the marker proteins CD9 and CD63 of exosomes were detected by Western blotting (WB). The expression of miRNA-21-5p in exosomes was determined by reverse transcription polymerase chain reaction (RT-PCR). The degree of lung tissue injury and fibrosis was observed by hematoxylin-eosin staining (HE) and Masson staining. The collagen content of lung tissue was detected by hydroxyproline (HYP) method. The expression of Smad7 protein in lung tissue was detected by WB. Results The results of pathological staining showed that compared with the control group, lung inflammatory cell infiltration, alveolar wall thickening, and collagen increase were observed after 4 weeks of dusting, and collagen deposition and silicon nodules appeared after 16 weeks of dusting. Compared with the control group, the expression level of HYP in the lung tissue of the quartz group was increased after 4 weeks and 16 weeks of dust staining (P<0.05). Transmission electron microscopy showed that exosomes were saucer-shaped, and the average particle size of exosomes was 95.8 nm by NTA. Positive expression of exosome marker proteins CD9 and CD81 was found by WB. Compared with the control group, the expression of exosome-derived miRNA-21-5p in alveolar lavage fluid in the quartz group increased in the 4th week and the 16th week (P<0.05), and the expression of Smad7 protein in lung tissue decreased (P<0.05). Conclusion Exosome-derived miRNA-21-5p and Smad7 may be involved in the mechanism of quartz dust-induced pulmonary fibrosis in rats.

Objective: To explore the effects of subchronic aluminum exposure on the level of N6-methyladenosine (m6A) methylation and the expression of N6-methyladenosine RNA binding protein 1 (YTHDF1) in the hippocampus of rats. Methods: Twenty-four healthy male SD rats were randomly divided into the control group (normal saline), the low dose group [10 μmol/kg Al(mal)3], the medium dose group [20 μmol/kg Al(mal)3] and the high dose group [40 μmol/kg Al(mal)3], with 6 rats in each group. The Al(mal)3 solution was administered via intraperitoneal injection on alternate days for 90 days. Escape latency, target quadrant dwell time and platform crossing times were tested to evaluate the learning and memory ability of the rats by the Morris water maze test after exposure. The brain tissue was weighted and the brain-to-body weight ratio was calculated after euthanasia. The level of m6A methylation and the expression of YTHDF1 were determined by enzyme-linked immunosorbent assay and western blot assay, respectively. Results: All rats survived during aluminum exposure period. The brain-to-body weight ratios of the control group and the low, medium and high dose groups were (0.46±0.06)%, (0.44±0.04)%, (0.49±0.06)% and (0.51±0.07)%, respectively, with no statistically significant differences (P>0.05). The escape latency of rats in the high dose group was longer than that in control and low group during the third to fifth day (both P>0.05). The escape latency of rats in all groups was shortened with the increase of training days (P<0.05). The target quadrant dwell time of rats in low, medium and high dose groups were lower than that in control group, and the platform crossing times of rats in high dose group were lower than that in control group (all P<0.05). The methylation level of m6A and expression level of YTHDF1 in hippocampus of rats in medium and high dose groups was higher than that in control group (both P<0.05). Conclusion The learning and memory impairment caused by subchronic aluminum exposure may be related to the increase of m6A methylation level and the decrease of YTHDF1 expression.

To investigate the alteration of c-Jun N-terminal kinase (JNK) activity after myocardial ischemia reperfusion injury (MIRI) and further explore the effect of naloxone postconditioning on MIRI. Forty male Sprague Dawley rats were randomly divided into five groups: sham operation (sham, n=8); ischemia reperfusion (IR, n=8); IR+naloxone 0.5 mg/kg (Nal L, n=8); IR+naloxone 1.0 mg/kg (Nal M, n=8); IR+naloxone 2.0 mg/kg (Nal H, n=8). Pathological changes of myocardial tissue were visualized by HE staining. The expression of p-JNK, and the apoptosis of cardiomyocytes were investigated with Western blotting and the TUNEL assay, respectively. Irregular arrangement and aberrant structure of myocardial fibers, cardiomyocytes with granular or vacuolar degeneration, and inflammatory cells infiltrating the myocardial interstitial regions characterized MIRI in the IR group. Signs of myocardial injury and inflammatory infiltration were less prominent in the Nal-treated groups. The expression of p-JNK in the sham group and in all Nal-treated groups was significantly lower than that in the IR group (p<0.01). The apoptosis index of cardiomyocytes in the IR group was significantly higher than in the sham group (p< 0.01). The apoptosis indices of cardiomyocytes in all Nal-treated groups were significantly reduced to 55.4%, 26.2%, and 27.6%, respectively, of the IR group (p< 0.01). This study revealed that Naloxone postconditioning before reperfusion inhibits p-JNK expression and decreases cell apoptosis, thus alleviating MIRI.
Animals ; Apoptosis ; Blotting, Western ; Humans ; In Situ Nick-End Labeling ; Ischemia ; JNK Mitogen-Activated Protein Kinases ; Male ; Myocardial Ischemia* ; Myocytes, Cardiac ; Naloxone* ; Rats* ; Rats, Sprague-Dawley ; Reperfusion Injury* ; Reperfusion*

Abstract: It is well-known that crystal form of a drug is a key factor impacting the physicochemical properties of the drug, which in turn affects its in vivo efficacy, safety and stability. The study on crystal forms of solid-state drugs is crucial for drug quality control, selection of production process and evaluation of clinical efficacy. The combination of chemometric and analytical techniques exhibited its great ability to analyze a large amount of multidimensional data, providing the possibility for quantification of trace amount of crystals (< 1%). Meanwhile, using the process analytical technology (PAT) to monitor the crystal content real-time during prescription preparation process can further realize the control on formulation quality and serve as a core technology to support the patent protection of crystalline forms. In this review, the combined application of crystal analytical techniques and chemometric methods for the quantitative analysis of trace crystals were summarized, aiming to provide guidance for the manufacturing of pharmaceutical preparations and their quality control.

Objective:To systematically review the association between delivery mode and exclusive breastfeeding rate during hospitalization and within the first six months of life.Methods:Observational studies on the association between delivery mode and feeding pattern were searched from PubMed, Web of Science, Cochrane Library, EBSCO, China Biomedical Literature Database, CNKI, Wanfang Database, and VIP Database from inception to October 2022. Two independent reviewers screened the literature, extracted data, and assessed the quality of included studies using Critical Appraisal Tools published by Joanna Briggs Institute or Newcastle-Ottawa Quality Scale (NOS). This meta-analysis was performed using R 4.1.0 software. Fixed-effect or random-effect models were used to pool data. Egger test and funnel plot were used to assess publication bias.Results:A total of 34 studies involving 597 203 subjects were included, including 22 cross-sectional studies and 12 cohort studies. All of the 22 cross-sectional studies were B-level quality, and eleven out of the 12 included cohort studies scored 7 points or above on the NOS scale with high quality. The results of meta-analysis showed that the likelihood of exclusive breastfeeding during hospitalization of women who had cesarean section was lower than those who delivered vaginally ( OR=0.33, 95% CI: 0.22-0.50, P<0.001); and so was the likelihood of exclusive breastfeeding at six months postpartum ( OR=0.61, 95% CI: 0.47-0.79, P<0.001). Conclusion:Current evidence suggests that cesarean section is a disadvantage to exclusive breastfeeding during hospitalization and within six months after delivery.

Objective:To systematically evaluate the clinical efficacy of oral ginger capsule or ginger powder in chemotherapy-induced nausea and vomiting in cancer patients.Methods:Computers searched Chinese Journal Full-text Database (CNKI), China Biomedical Literature Database (CBM), Wanfang Database, PubMed, EMbase, Web of Science, and Cochrane Library about oral chemotherapy in patients with cancer ginger correlation clinical curative effect of nausea and vomiting randomized controlled trial, supplemented by other search methods, the time range was built until July 2019. Quality evaluation and data extraction were performed independently by two investigators, and Meta analysis was performed by RevMan5.3 software.Results:A total of 12 articles and 13 studies were included, with a total of 1 105 patients. Meta-analysis showed that oral ginger capsule or ginger powder reduced the incidence of acute vomiting (risk ratio value was 0.76, 95% confidence interval was 0.59-0.98, P<0.05) and the severity of vomiting (mean difference value was-0.79, 95% confidence interval was-1.36--0.23, P<0.01), including the severity of acute vomiting (mean difference value was-1.39, 95% confidence interval was-2.72--0.06, P<0.05) and the severity of delayed vomiting (mean difference value was-0.46, 95% confidence interval was-0.82--0.10, P<0.05). However, there was no significant difference between the two groups in the incidence and severity of acute and delayed nausea ( P>0.05). Conclusions:This study demonstrates that oral ginger capsule or ginger powder is a complementary treatment for chemotherapy-induced nausea and vomiting in cancer patients, and more high-quality studies are needed to validate its clinical efficacy in the future.

Objective:To develop a verification platform based on Monte Carlo (MC) for independent dose verification of volumetric modulated arc therapy (VMAT) plans.Methods:The head model including collimator of Varian TrueBeam linear accelerator was constructed by using EGSnrc/BEAMnrc, and the independent dose verification platform for the patients’ VMAT plans was built based on the head model and an in-house code. The percent depth dose (PDD) curves and off-axis ratios for different field sizes, the dose distribution of two irregular fields and three VMAT plans of the head and neck, chest, and pelvis were simulated using the platform. The simulated results of the PDD curves and the off-axis ratios of different field sizes were compared with the blue water measurement results. The difference between the irregular fields and the actual ArcCHECK measurements was also investigated. Besides, the differences among the MC simulated dose, TPS calculated dose and the ArcCHECK measured dose were analyzed by several methods, such as γ analysis and dose-volume histogram to verify whether the platform could be independently employed for dose verification.Results:The MC simulated results of PDD curves and off-axis ratios from 4 cm×4 cm to 40 cm×40 cm were in good agreement with the measured results. And the γ passing rates between the MC simulation and the ArcCHECK measurement for the irregular fields were above 98.1% and 99.1% for 3%/2 mm and 3%/3 mm, respectively. For VMAT plans of three patients, the γ results between the MC simulated dose and ArcCHECK measured dose were better than 93.8% and 95.9% under the criteria of 3%/2 mm and 3%/3 mm respectively. At the same time, the γ passing rates of nasopharyngeal, lung, and rectal cancers were 95.2%, 98.6% and 98.9% based on 3D γ analysis using TPS calculated dose and MC simulated dose under the criteria of 3%/3 mm; the passing rates of these three were 90.3%, 95.1% and 96.7% for 3%/2 mm, respectively.Conclusions:The simulation results of the MC-based verification platform developed in this study show a good agreement with the actual measurement results, and the simulation results are closer to the real dose distribution using the patients’ data. The preliminary results demonstrate that the platform can be used for accurate independent dose verification of VMAT plans.