Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing. Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing. Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.

Aim To evaluate the effectiveness and safety of remimazolam tosylate for administering general anesthesia in morbidly obese patients.Methods This clinical trial was conducted at a single center from De-cember 2021 to October 2023.It assessed 108 morbid-ly obese patients(body mass index,BMI≥40)who underwent laparoscopic sleeve gastrectomy.Patients were randomly assigned to either the remimazaolam group(Group R)or the propofol group(Group P)for general anesthesia induction and maintenance.The primary outcome was to compare the incidence of ad-verse events and postoperative recovery characteristics between the two groups.Results During induction pe-riod,the incidence of adverse events was higher in group P,including hypotension(P＜0.01),hypox-emia(P＜0.05),bradycardia(P＜0.01),and in-creased vasopressor requirement(P＜0.05).The time to loss of consciousness and BIS falling to 60 was shor-ter in group P than in group R(P＜0.01).There were no statistically significant differences between the two groups in terms of postoperative quality of recovery(QoR-40 score),24-hour postoperative pain visual an-alogue scale(VAS)scores and morphine consump-tion.In conclusion,remimazolam tosylate,utilized for anesthesia induction in morbidly obese patients,signif-icantly reduced hypotension and hypoxemia compared to propofol,while it could also maintain similar postop-erative recovery quality.Conclusions Remimazolam is effective in reducing the incidence of hypotension and hypoxaemia during the induction period of general anaesthesia in morbidly obese patients and it is compa-rable to propofol in terms of quality of postoperative re-covery.

Traumatic brain injury ( TBI ) has caused serious economic and social burdens, but due to its heterogeneity, there is no effective treatment. In TBI with different severity, diffuse axonal injury (DAI) incidenceis high. The investigation on DAI will contribute to the diagnosis and treatment of TBI. In this study, the classification of TBI and the research status of DAI were summarized. The method to judge the severity of TBI and DAI, and animal experimental models and related injury criteria and thresholds were reviewed. The result show that DAI is mainly generated by rotational acceleration and it is related to angular acceleration, angular velocity and duration. Several TBI animal models can induce the pathology of DAI, and inertial rotation models which can produce only rotational acceleration have been developed. However, these models are instantaneous rotation models, and the rotation duration is uncontrollable, thus a longer duration is impossible, and DAI severity under long rotational motion cannot be studied. The study proposes that a new rotation animal model which can control rotation duration should be developed. The development of the animal model and investigation on pathomechanism of the model will contribute to the prevention and treatment of DAI.

OBJECTIVE To esta blish the method for simultaneous determination of 25 components （such as berberine ， magnoflorine and hydroxysafflor yellow A ）in Bawei xiaobopi capsules. METHODS High-performance liquid chromatography- tandem triple quadrupole mass spectrometry （HPLC-QqQ-MS）method was adopted. The determination was performed on WondaSil C18-WR column with mobile phase consisted of 0.1％ formic acid solution-methanol （gradient elution ）at the flow rate of 0.5 mL/min. The column temperature was 25 ℃，and sample size was 5 μL. Electrospray ionization source was scanned in positive and negative ion mode at the same time ，with multiple reaction monitoring. The capillary voltage was 4 000 V（+）and 2 500 V（－）. The drying gas flow rate was 11 L/min with the temperature of 300 ℃. The pressure was 15 psi. RESULTS Totally 25 components of Bawei xiaobopi capsules had good linear relationship within a certain range ，such as magnolflorine ，jatrorrhizine，berberine， palmatine，bufotenine，bufotenidine，piperine，glycyrrhizic acid ，ferulic acid ，ferulic acid 4-O-β-D-glucopyranoside，hydroxysafflor yellow A ，chlorogenic acid ，gallic acid ，chebulagic acid ，corilagin，ellagic acid ，liquiritigenin，liquiritin，rutin，quercetin， glycocholic acid ，cholic acid ，glycochenodeoxycholic acid ，glycodeoxycholic acid ，ursodeoxycholic acid （r≥0.999 0）. The limits of quantitation were 0.62-554.50 ng/mL；the limits of detection were 0.18-166.30 ng/mL.RSDs of precision ，repeatability and stability（24 h）tests were all lower than 3.00％. The recovery rates were 80％-115％（all RSDs lower than 3.00％，n＝6）. The contents of above 25 components were 16.94-20.82，3.78-5.17，9.11-11.43，0.24-0.30，0.20-0.39，0.74-1.16，0.79-0.89，3.26-3.35， 0.48-0.66，11.96-13.35，2.30-3.12，0.19-0.21，6.07-8.83，10.42-10.48，1.43-1.64，4.17-4.76，0.14-0.15，0.46-0.52，0.04，0.01， 0.59-0.63，0.20-0.23，0.02，0.15-0.16，0.01 mg/g，respectively. CONCLUSIONS Established method is simple ，sensitive and stable，and can be used for content determination of 25 components in Bawei xiaobopi capsules simultaneously.

Animals ; Biological Assay ; Nucleic Acid Amplification Techniques ; Phlebovirus/isolation & purification* ; Psychodidae/virology* ; Recombinases/metabolism* ; Reverse Transcription

OBJECTIVE: To compare the maternal and infant outcomes of pregnant women infected with human immunodeficiency virus(HIV)treated with different regimens of highly active antiretroviral therapy(HAART).METHODS: For pregnant women infected with the human immunodeficiency virus(HIV)who received antiviral therapy and delivered in the Eighth Peple's Hospital of Guangzhu between May 2015 and June 2018,they will be grouped according to different treatment options. The pregnant women's body weight,CD4+T lymphocytes,white blood cells,hemoglobin,serum albumin,neonatal body weight and adverse pregnancy outcomes were compared and analyzed.RESULTS:(1)There was no significantly statistical difference between the two groups of pregnant women in terms of body weight,white blood cells,hemoglobin or serum albumin(P>0.05).(2)The changes of CD4+T lymphocytes in the two groups of pregnant women before and after treatment were statistically different(P<0.05).(3)There was no significant difference in neonatal weight,gestational age or Apgar score between the two groups(P>0.05).(4)There was no significantly statistical difference in the incidence of premature birth,premature rupture of fetal membrane,low birth weight,low amniotic fluid,fetal malformation or neonatal asphyxia between the two groups(P>0.05).Until December 2018,there were no positive reports of HIVRNA and HIV antibody detection in two groups of infants.CONCLUSION: The two HAART schemes have no significant difference in the influence on nutritional status,immune status or maternal and infant outcomes of HIV-infected pregnant women,and they are both effective and feasible,and vertical transmission of HIV from mother to child can be blocked.

Objective To evaluate the practicability of using CRISPR/Cas9 genome editing tech-nology for inhibition of hepatitis B virus ( HBV) replication. Methods Two sgRNA targeting sites were de-signed for the S region of HBV genome. The CRISPR/Cas9 expression plasmids specific for HBV were con-structed and then transfected into a cell line expressing HBV genome(HepG2-N10). The cytotoxicity of cells transfected with different expression plasmids were detected by MTT assay. The levels of hepatitis B surface antigen ( HBsAg ) were determined by using chemiluminescent immunoassay ( CLIA ) . The expression of HBV at mRNA level was analyzed by quantitative real-time PCR ( qRT-PCR) . The qPCR was performed for the detection of extracellular and intracellular HBV DNA. The next-generation sequencing ( NGS) Illumina MiSeq Platform was used to analyze HBV genome editing. Results No significant cytotoxic effects were de-tected in HepG2-N10 cells transfected with different expression plasmids. Compared with the cells carrying pCas-Guide-GFP-Scramble, the levels of HBsAg in the supernatants of transfected cell culture harboring pCas-Guide-GFP-G1 and pCas-Guide-GFP-G2 were decreased by 24. 2% (P<0. 05) and 16. 9% (P>0. 05), respectively. The levels of HBsAg in cells transfected with pCas-Guide-GFP-G1 and pCas-Guide-GFP-G2 were respectively decreased by 16. 4% (P>0. 05) and 32. 1% (P>0. 05) as compared with that of pCas-Guide-GFP-Scramble transfected group. The expression of HBV at mRNA level was inhibited as indica-ted by the results of qRT-PCR. Moreover, the levels of extracellular HBV DNA were respectively suppressed by 23% (P>0. 05) and 35% (P<0. 05), and the levels of intracellular HBV DNA were respectively sup-pressed by 7. 2% (P>0. 05) and 18% (P>0. 05). Different types of insertion/deletion mutation were de-tected in HBV genome by high-throughput sequencing. Conclusion HBV-specific CRISPR/Cas9 system could inhibit the expression of HBV gene and the replication of virus. Therefore, the CRISPR/Cas9 genome editing technology might be used as a potential tool for the treatment of persistent HBV infection.

Non-progressive congenital myopathy is a group of muscle diseases occurring at birth or during teenage years. A number of new reports of congenital myopathy, such as homogeneous bodies myopathy, muscle quality control myopathy and type 1 fiber predominance have recently been reported, but they lack of sufficient quantity and constant clinico-pathologic manifestations. This paper reports two cases of congenital myopathy with type 1 fiber predominance confirmed by muscle biopsy. The clinical manifestations of the two children (a 4.5-year-old girl and an 11-year-old boy) included non-progressive symptoms of muscle weakness, skeletal deformities and other clinical features of congenital myopathy. The physical examinations showed a long face or figure and funnel chest or kyphosis/scoliosis, high palatal arch and wing-like shoulder. Serum levels of creatine kinase were normal but slightly elevated serum lactate dehydrogenase levels were noted in the two children. The skeletal muscle biopsy by ATPase staining showed that type 1 fibers accounted for more than 90% of the total number of muscle fibers. No other abnormal pathological changes, such as central cores, muscle tube and central nuclei, were found in the two children.
Diagnosis, Differential ; Female ; Humans ; Infant ; Male ; Muscle, Skeletal ; pathology ; Myopathies, Structural, Congenital ; diagnosis ; pathology ; therapy

OBJECTIVETo investigate the clinical characteristics of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) patients in China.

METHODSTotally 143 HIV/AIDS patients who were first diagnosed in Peking Union Medical College Hospital form January 1988 to April 2006 were enrolled in this study. Clinical characteristics were retrospectively analyzed.

RESULTSAmong 143 HIV/ AIDS patients, 57 patients had no clinical symptoms and were confirmed by routine examinations; 86 patients had clinical symptoms, including fever (n = 50), weight loss (n = 18), and discomforts involving respiratory system (n = 34), gastrointestinal system (n = 16), and derma and mucosa (n = 17). Opportunistic infections (OIs) such as pneumocystis jiroveci pneumonia (PCP) (n = 27), oropharyngeal candidiasis (n = 16), tuberculosis (n = 15) , and cytomegalovirus (CMV) infection (n = 9) were also observed in patients whose CD4 + T cell counts were less than 200/mm3. Most CMV infection and cryptococcal meningitis occurred in patients whose CD4 + T cell counts were less than 100/mm3. CD4 + T cell count was negatively correlated with plasma viral load (r = -0.420, P = 0.001).

CONCLUSIONSFever, dyspnea, and weight loss are the most common symptoms in the patients of this study. The respiratory system, gastrointestinal system, derma and mucosa are the most commonly affected areas by OIs, and PCP is the most common OI. The occurrence of OIs corelates with CD4 + T cell count.

AIDS-Related Opportunistic Infections ; immunology ; Acquired Immunodeficiency Syndrome ; complications ; Adolescent ; Adult ; Aged ; CD4 Lymphocyte Count ; China ; Dyspnea ; etiology ; Emaciation ; etiology ; Female ; Fever ; etiology ; HIV Infections ; complications ; Humans ; Male ; Middle Aged ; Pneumonia, Pneumocystis ; immunology ; Retrospective Studies