Lipopolysaccharides (LPS) are major outer membrane components of Gram-negative bacteria. Unlike most Gram-negative bacteria, Acinetobacter baumannii can still survive and acquire polymyxin resistance after complete loss of LPS. Previous studies of LPS-deficient Acinetobacter baumannii mostly focused on LPS-deficient strains induced by polymyxin, whose background was complex and unstable. To investigate LPS loss-mediated polymyxin resistant-Acinetobacter baumannii, this study constructed a stable and clear background LPS-deficient strain by knocking out lpxC gene in Acinetobacter baumannii ATCC 19606 with CIRSPR/Cas9, and then studied the phenotypic changes of the lpxC-deficient strain including morphology, growth rate, antibiotic susceptibility, virulence, membrane permeability, and membrane potential. Animal experiments were approved by the Animal Care and Welfare Committee Institute of Medicinal Biotechnology, CAMS and PUMC (approval number: IMB-20240119D₉). The results indicated that the lpxC gene of Acinetobacter baumannii ATCC 19606 was successfully knocked out. After losing the lpxC, the strain underwent the morphological change from rod-shaped to spherical. Furthermore, it leads to reduced growth rate, enhanced membrane permeability, decreased membrane potential, lower virulence, and increased antibiotic susceptibility to β-lactams, quinolones, aminoglycosides, macrolides, glycopeptides. The lpxC deletion results in significant changes in membrane homeostasis and adaptability of Acinetobacter baumannii. Understanding the phenotypic changes of colistin-resistant Acinetobacter baumannii mediated by LPS loss is useful for exploring the resistance mechanism of Acinetobacter baumannii and developing new therapeutic strategies.

The modernization and development of traditional Chinese medicine has led to higher standards for the quality of traditional Chinese medicine products. The extraction process is a crucial component of traditional Chinese medicine production, and it directly impacts the final quality of the product. However, the currently relied upon methods for quality assurance of the extraction process, such as simple wet chemical analysis, have several limitations, including time consumption and labor intensity, and do not offer precise control of the extraction process. As a result, there is significant value in incorporating near-infrared spectroscopy (NIRS) in the production process of traditional Chinese medicine to improve the quality control of the final products. In this study, we focused on the extraction process of Xiao'er Xiaoji Zhike oral liquid (XXZOL), using near-infrared spectra collected by both a Fourier transform near-infrared spectrometer and a portable near-infrared spectrometer. We used the concentration of synephrine, a quality control index component specified by the pharmacopoeia, to achieve rapid and accurate detection in the extraction process. Moreover, we developed a model transfer method to facilitate the transfer of models between the two types of near-infrared spectrometers (analytical grade and portable), thus resolving the low resolution, poor performance, and insufficient prediction accuracy issues of portable instruments. Our findings enable the rapid screening and quality analysis of XXZOL onsite, which is significant for quality monitoring during the traditional Chinese medicine production process.

In recent years, researchers, pharmaceutical companies, and political makers gradually using more real-world data (RWD) to produce real-world evidence (RWE) for policy-making. A research team of Harvard University launched the RCT DUPLICATE project in 2018, aiming to replicate 30 randomized controlled trials using the medical claims database in order to explore methods for quantifying the efficacy-effectiveness gap and explain its potential sources, to enhance the credibility of the RWE. This paper reviews the background of RCT DUPLICATE Initiative, highlights the research purposes, research design and implementation process of the RCT DUPLICATE Initiative, to help domestic scholars better understand the scope and application value of RWE.
Humans ; Randomized Controlled Trials as Topic ; Cognition ; Databases, Factual ; Research Personnel ; Universities

With the promotion and application of big medical data, non-interventional real-world evidence (RWE) has been used by regulators to assess the effectiveness of medical products. This paper briefly introduces the latest progress and research results of the RCT DUPLICATE Initiative launched by the research team of Harvard University in 2018 and summarizes relevant research experience based on the characteristics of China's medical service to provide inspiration and reference for domestic scholars to conduct related RWE research in the future.
Humans ; Randomized Controlled Trials as Topic ; Cognition ; Big Data ; Universities

Objective: To explore the feasibility of preoperative designing for anterolateral femoral flap using three longitudinal and five transversal strategy. Methods: From September 2015 to January 2017, 71 patients (73 pieces) were treated using anterolateral thigh flap in the Affiliated Hospital of Zunyi Medical University. There were 48 males and 23 females, with the age of (36±18) years. The three longitudinal and five transversal designing was performed before operation. Portable Doppler ultrasound was used to detect the piercing-out position (P point) of perforators in Ⅰ, Ⅱ, Ⅲ, Ⅳ areas. The diameter, direction and length (lower subcutaneous segment of perforators) of perforators after leaving piercing-out position were observed during the operation. The piercing-in positions on superficial fascia and the dermis were observed. Results: The data of 71 adults (73 legs) were evaluated. Before and during the operation, all the perforations were found in Ⅰ, Ⅱ, Ⅲ and Ⅳ areas. The number of perforators in region Ⅰ was 1.32±0.33, with the diameter of perforator at the piercing-out position was (0.79±0.13) mm. The length of subfascial segment of perforators was (1.54±0.97) cm, and the direction was oblique superolateral. The number of perforators in Ⅱ region was 1.21±0.53, with the diameter of perforator at the piercing-out position was (0.63±0.13) mm. The length of subfascial segment of perforators was (2.25±0.54) cm, and the direction was oblique inferoanterior. The distance form piercing-in position to line L was (0.84±0.42) cm. The number of perforators in the Ⅲ area was 2.22±0.49, with the diameter of perforators at the piercing-out position was (0.53±0.12) mm. The length of subfascial segment of perforator was (1.96±0.44) cm, and the direction was oblique inferoanterior. The distance between piercing-in positions to line L was (0.74±0.51) cm. The number of perforators in region Ⅳ was 1.41±0.72, with the length of subfascial segment of perforators was (1.22±0.45) cm and the direction was oblique inferolateral. There were 27 cases with oblique perforators, accounting for 37% of the total number of thighs. Conclusions This three longitudinal and five transversal designing is helpful to detect the expenditure point in the anterolateral femoral region, and is assistant to skin flap designing.

Objective To explore the effect of remind-to-move treatment on upper limb motor function,activities of daily living and participation in patients with subacute stroke. Methods From February,2016 to October,2017,45 patients with mild to medium upper limbs dysfunction after stroke were randomly assigned to control group(n=23)and experimental group(n=22).The control group accepted rou-tine occupational therapy,while the experimental group wore a wristwatch on the hemiplegic forearm to encour-age the predetermined training programs,for three weeks.They were measured with Fugl-Meyer Assessment-Up-per Extremity(FMA-UE),Function Independence Measurement(FIM),Motor Activity Log(MAL),and Stroke Impact Scale(SIS)before and after treatment. Results Both groups improved in part of the scores of three scales after treatment(P<0.05),and improved more in the experimental group than in the control group in scores of FMA-UE and FIM,and some sub-scores of MAL and SIS(t>1.183,P<0.05),with no significant difference in other indexes(P>0.05). Conclusion Remind-to-move treatment can promote the recovery of upper limb motor function,activities of daily living and participation in the patients with subacute stroke.

As an important part of epigenetic marker, DNA methylation involves in the gene regulation and attracts a wide spread attention in biological auxology, geratology and oncology fields. In forensic science, because of the relative stable, heritable, abundant, and age-related characteristics, DNA methylation is considered to be a useful complement to the classic genetic markers for age-prediction, tissue-identification, and monozygotic twins' discrimination. Various methods for DNA methylation detection have been validated based on methylation sensitive restriction endonuclease, bisulfite modification and methylation-CpG binding protein. In recent years, it is reported that the third generation sequencing method can be used to detect DNA methylation. This paper aims to make a review on the detection method of DNA methylation and its applications in forensic science.
CpG Islands ; DNA Methylation/genetics* ; Epigenesis, Genetic ; Epigenomics ; Forensic Genetics/trends* ; Genetic Markers/genetics* ; Humans ; Sulfites ; Twins, Monozygotic/genetics*

ObjectiveTo evaluate the effect of ejaculatory duct dilation combined with seminal vesicle clysis in the treatment of refractory hematospermia.

METHODSUsing ureteroscopy, we treated 32 patients with refractory hematospermia by transurethral dilation of the ejaculatory duct combined with clysis of the seminal vesicle with diluent gentamicin.

RESULTSThe operation was successfully accomplished in 31 cases, with the mean operation time of 32 (26－47) minutes. The patients were followed up for 6－39 (mean 23.6) months. No complications, such as urinary incontinence and retrograde ejaculation, were found after operation. Hematospermia completely disappeared in 27 cases, was relieved in 1, and recurred in 3 after 3 months postoperatively. Those with erectile dysfunction or mental anxiety symptoms showed significantly decreased scores of IIEF-Erectile Function (IIEF-EF) and Self-Rating Anxiety Scale (SAS).

CONCLUSIONSEjaculatory duct dilation combined with seminal vesicle clysis under the ureteroscope, with its the advantages of high effectiveness and safety, minimal invasiveness, few complications, and easy operation, deserves general clinical application in the treatment of refractory hematospermia.

Dilatation ; Ejaculatory Ducts ; surgery ; Genital Diseases, Male ; Hemospermia ; surgery ; Humans ; Male ; Postoperative Period ; Recurrence ; Seminal Vesicles ; surgery ; Ureteroscopy

In order to develop a rapid detection kit for novel avian influenza virus (AIV) subtype H7N9, two sets of specific primers and probes were designed based on the nucleotide sequences of hemagglutinin antigen (HA) and neuraminidase antigen (NA) of novel H7N9 virus (2013) available in GenBank to establish the method of TaqMan probe-based multiplex real-time RT-PCR for rapid detection of AIV subtype H7N9. The primer and probe of HA were for all H7 subtype AIVs, while the primer and probe of NA were only for novel N9 subtype AIVs. The results showed that this method had high sensitivity and specificity. This method was applicable to the testing of positive standard sample with a minimum concentration of 10 copies/microL; it not only distinguished H7 subtype from H1, H3, H5, H6, and H9 subtypes, but also distinguished novel N9 subtype from traditional N9 subtype. A total of 2700 samples from Zhuhai, China were tested by this method, and the results were as expected. For the advantages of sensitivity and specificity, the method holds promise for wide application.
Animals ; Birds ; virology ; Influenza A Virus, H7N9 Subtype ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; prevention & control ; virology ; Real-Time Polymerase Chain Reaction ; methods ; Species Specificity ; Taq Polymerase ; metabolism ; Time Factors

In order to study the proliferation inhibition effect of H5N1 subtype avian influenza virus (AIV) with small interfere RNA (siRNA), a total of 4 siRNAs were designed in accordance with the NP and PA genes of H5N1 subtype AIV, the siRNAs were then transfected to chicken embryo fibroblast(CEF), CEF was infected with H5N1 subtype AIV after 6 hrs. Virus titer of cell supernatant was tested at 16-56hrs post infection, and pathological changes of the cells was observed; mRNA levels of NP, PA, HA and p13-actin gene were tested at 36hrs post infection. The results showed that these 4 siRNAs could inhibit the prolif-eration of H5N1 subtype AIV in CEF in varying degrees, and one siRNA targeting PA was best per-formed. The experimental results also showed that the inhibition effect was decreased with the time prolonged. This research provides a basis for further studying RNAi on AIV prevention and control.
Actins ; genetics ; Animals ; Chick Embryo ; DNA Primers ; genetics ; Fibroblasts ; virology ; Hemagglutination ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Hemagglutinins ; genetics ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; growth & development ; physiology ; RNA Interference ; RNA Replicase ; genetics ; RNA, Small Interfering ; chemical synthesis ; genetics ; RNA-Binding Proteins ; genetics ; Real-Time Polymerase Chain Reaction ; Specific Pathogen-Free Organisms ; Transfection ; Viral Core Proteins ; genetics ; Viral Proteins ; genetics ; Virus Replication