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.

Toxic Chinese materia medica has been highly valued by its specialized and effective effects， but its safe application has become an urgent clinical problem to be solved. Compatibility for toxic attenuation is an important method for the rational clinical application of toxic Chinese materia medica as well as the promotion of its therapeutic advantages. The theory of “heterogeneous medicinals mutual restriction” elaborated in this article has been formed through long-term clinical practice and cognition， and refinement of clinical experience， which means that the nature partiality of toxic Chinese materia medica can be adjusted， and the toxicity can be suppressed through reasonable combination with herbal medicinalsof different properties， flavors， and effects. This theory covers the modes of compatibility for toxicity attenuation and the interaction relationships， like the restriction of medicinals with different properties and flavors， restriction of medicinals with different effects， and inhibiting toxins by reinforcing healthy qi. The opposite and complementary effects of various medicinals combinations are an extension of the connotations of this theory， and the principles can be explained from material basis and mechanism of action. Under the guidance of this theory， it is possible to optimize the compound prescription strategies of toxic Chinese materia medica， and provide new strategies for the clinical combinations of toxic Chinese materia medica， thereby achieving the reduction of toxicity and enhancement of effectiveness of the compound formulas.

Objective To investigate the effect and mechanism of rhubarb Tangluo pill on liver injury in type 2 diabetic rats.Methods ZDF(fa/fa)rats were given high-fat diet to induce type 2 diabetes model,and were randomly divided into model group,positive control group(0.18 g·kg-1 metformin)and experimental-L,-M,-H groups(0.54,1.08 and 2.16 g·kg-1 rhubarb Tangluo pill),with 8 rats in each group.Eight ZDF(fa/+)rats were selected as control group.The control group and model group were given equal volume of pure water once a day for 12 weeks.An oral glucose tolerance test(OGTT)was performed after administration.Fasting blood glucose level,body mass and liver mass of rats were measured and liver index was calculated.The contents of glutamic-pyruvic transaminase(GPT),glutamic oxalacetic transaminase(GOT),triglyceride(TG)and total cholesterol(TC)in serum were detected.The histomorphologic changes of liver were observed by hematoxylin-eosin(HE)staining and Masson staining.The protein expression of phosphorylated insulin receptor substrate 1(p-IRS1),phosphorylated protein kinase B(p-Akt)and glucose transporter 4(GLUT4)were detected by Western blotting.Results After administration,the fasting blood glucose levels of control group,model group,positive control group and experimental-H group were(4.71±0.45),(29.9±2.97),(15.28±4.52)and(13.84±1.55)mmol·L-1,respectively;the liver index were 2.31±0.46,4.03±0.18,3.37±0.23 and 3.38±0.24;the relative expression level of p-IRS1 protein were 1.00±0.36,4.00±0.11,1.62±0.27 and 1.90±0.17,respectively;the relative expression levels of p-Akt protein were 1.00±0.25,0.21±0.04,0.73±0.15 and 0.54±0.04,respectively;GLUT4 protein relative expression levels were 1.00±0.11,0.40±0.08,0.86±0.04 and 0.70±0.06,respectively.Compared with the model group,the above indexes in the experimental-H group were statistically significant(P＜0.01,P＜0.05).Conclusion Rhubarb Tanglu pill can effectively improve glycolipid metabolism and liver injury in type 2 diabetes mellitus,and its mechanism may be related to the activation of IRS1/Akt signaling pathway.

OBJECTIVE: To evaluate the safety and efficacy of catheter-directed thrombolysis (CDT) versus systemic thrombolysis (ST) in the treatment of pulmonary embolism (PE). METHODS: The Cochrane Library, PubMed, and Embase databases were searched to collect the literature on the comparison of the results of CDT and ST in the treatment of PE from the beginning of their records to May 2020, and meta-analysis was performed by STATA software (version 15.1). Using standardized data-collection forms, the authors screened the studies and independently extracted data, and assessed the quality of the studies using the Newcastle-Ottawa Scale for cohort studies. Cohort studies that examined the following results were included in the current study: in-hospital mortality, all-cause bleeding rate, gastrointestinal bleeding rate, intracranial hemorrhage rate, the incidence of shock, and hospital length of stay. RESULTS: A total of eight articles, with 13,242 participants, involving 3962 participants in the CDT group and 9280 participants in the ST group were included. CDT compared with ST in the treatment of PE can significantly affect in-hospital mortality rate [odds ratio (OR) = 0.41, 95% CI: 0.30-0.56, P < 0.05], all-cause bleeding rate (OR = 1.20, 95% CI: 1.04-1.39, P = 0.012), gastrointestinal bleeding rate (OR = 1.43, 95% CI: 1.13-1.81, P = 0.003), the incidence of shock (OR = 0.46, 95% CI: 0.37-0.57, P < 0.05), and hospital length of stay [standard mean difference (SMD) = 0.16, 95% CI: 0.07-0.25, P < 0.05]. However, there was no significant effect on intracranial hemorrhage rate in patients with PE (OR = 0.70, 95% CI: 0.47-1.03, P = 0.070). CONCLUSIONS CDT is a viable alternative to ST in the treatment of PE, as it can significantly reduce in-hospital mortality rate, all-cause bleeding rate, gastrointestinal bleeding rate, and incidence of shock. However, CDT may prolong hospital length of stay to a certain extent. Further research is needed to evaluate the safety and efficacy of CDT and ST in the treatment of acute PE and other clinical outcomes.

OBJECTIVE: To examine whether the combination of Naoxintong Capsule with standard care could further reduce the recurrence of ischemic stroke without increasing the risk of severe bleeding. METHODS: A total of 23 Chinese medical centers participated in this trial. Adult patients with a history of ischemic stroke were randomly assigned in a 1:1 ratio using a block design to receive either Naoxintong Capsule (1.2 g orally, twice a day) or placebo in addition to standard care. The primary endpoint was recurrence of ischemic stroke within 2 years. Secondary outcomes included myocardial infarction, death due to recurrent ischemic stroke, and all-cause mortality. The safety of drugs was monitored. Results were analyzed using the intention-to-treat principle. RESULTS: A total of 2,200 patients were enrolled from March 2015 to March 2016, of whom 143 and 158 in the Naoxintong and placebo groups were lost to follow-up, respectively. Compared with the placebo group, the recurrence rate of ischemic stroke within 2 years was significantly lower in the Naoxintong group [6.5% vs. 9.5%, hazard ratio (HR): 0.665, 95% confidence interval (CI): 0.492-0.899, P=0.008]. The two groups showed no significant differences in the secondary outcomes and safety, including rates of severe hemorrhage, cerebral hemorrhage and subarachnoid hemorrhage (P>0.05). CONCLUSION The combination of Naoxintong Capsule with standard care reduced the 2-year stroke recurrence rate in patients with ischemic stroke without increasing the risk of severe hemorrhage in high-risk patients. (Trial registration No. NCT02334969).
Adult ; Humans ; Secondary Prevention/methods* ; Ischemic Stroke ; Stroke/prevention & control* ; Cerebral Hemorrhage/complications* ; Double-Blind Method ; Platelet Aggregation Inhibitors

OBJECTIVE: To explore the differences in the factors associated with endometriosis between Chinese and British patients. METHODS: This case-control study was conducted in 387 patients with endometriosis and 199 non-endometriosis patients admitted to John Radcliffe Hospital (Oxford, UK) and in 101 patients with endometriosis and 50 non-endometriosis patients admitted in the First Affiliated Hospital of Guangzhou University of Chinese Medicine. The clinical data including height, weight, body mass index, marital status, employment, menstruation, fertility, and operation reasons were collected via a standardized WERF EPHect questionnaire. RESULTS: Multivariate logistic regression analysis indicated that body mass index, surgery for dysmenorrhea, history of pregnancy, counts of previous surgeries for endometriosis and status of employment were all significantly associated with endometriosis in the UK (P < 0.05), while a history of dysmenorrhea was significantly correlated with endometriosis in Chinese patients (P < 0.05). CONCLUSION Dysmenorrhea may be the most important common factor associated with endometriosis in China and the UK, but the other factors contributing to endometriosis may differ between these two countries.
Case-Control Studies ; Dysmenorrhea/complications* ; Endometriosis/complications* ; Female ; Humans ; Menstruation ; Pregnancy ; United Kingdom

OBJECTIVE: To investigate the therapeutic effect of Epothilone D on traumatic optic neuropathy (TON) in rats. METHODS: Forty-two SD rats were randomized to receive intraperitoneal injection of 1.0 mg/kg Epothilone D or DMSO (control) every 3 days until day 28, and rat models of TON were established on the second day after the first administration. On days 3, 7, and 28, examination of flash visual evoked potentials (FVEP), immunofluorescence staining and Western blotting were performed to examine the visual pathway features, number of retinal ganglion cells (RGCs), GAP43 expression level in damaged axons, and changes of Tau and pTau-396/404 in the retina and optic nerve. RESULTS: In Epothilone D treatment group, RGC loss rate was significantly decreased by 19.12% (P=0.032) on day 3 and by 22.67% (P=0.042) on day 28 as compared with the rats in the control group, but FVEP examination failed to show physiological improvement in the visual pathway on day 28 in terms of the relative latency of N2 wave (P=0.236) and relative amplitude attenuation of P2-N2 wave (P=0.441). The total Tau content in the retina of the treatment group was significantly increased compared with that in the control group on day 3 (P < 0.001), showing a consistent change with ptau-396/404 level. In the optic nerve axons, the total Tau level in the treatment group was significantly lower than that in the control group on day 7 (P=0.002), but the changes of the total Tau and pTau-396/404 level did not show an obvious correlation. Epothilone D induced persistent expression of GAP43 in the damaged axons, detectable even on day 28 of the experiment. CONCLUSION Epothilone D treatment can protect against TON in rats by promoting the survival of injured RGCs, enhancing Tau content in the surviving RGCs, reducing Tau accumulation in injured axons, and stimulating sustained regeneration of axons.
Animals ; Disease Models, Animal ; Epothilones ; Evoked Potentials, Visual ; Nerve Regeneration/physiology* ; Optic Nerve Injuries/metabolism* ; Rats ; Rats, Sprague-Dawley ; Retinal Ganglion Cells/physiology*

Objective:To investigate the postoperative changes in levels of 25-hydroxy vitamin D (25-[OH]D], parathyroid hormone (PTH) and other relevant biomarkers in obese patients receiving metabolic surgery and analyze the dynamic changes in relevant biomarkers in the short term (after 3-6 months) and the long term (after 12-24 months).Methods:A total of 96 obese patients who underwent metabolic surgery and received follow-up examinations from January 2018 to January 2020 were included. Baseline and postoperative data were collected, including anthropometric data (height, weight, neck circumference, waistline and hipline) and laboratory test results (fasting glucose, glycated hemoglobin, 25-[OH)D, PTH, serum calcium and serum phosphorus). Body mass index (BMI) and waist-hip ratio were calculated. The anthropometric data were analyzed by repeated measures analysis of variance and laboratory data were compared between groups using t test and Kruskal-Wallis test. Results:96 patients (33 males and 63 females) were included, of whom 49 were complicated with diabetes. Prior to surgery, 79 (79.17%) of the patients had 25-(OH)D deficiency (< 20 μg/L), 16 (16.67%) had 25-(OH)D insufficiency (≥ 20 μg/L and < 30 μg/L) and 23 (23.96%) had high PTH levels (> 70 ng/L). After the surgery, 25-(OH)D level was transiently increased in the short term ( P = 0.01) but declined thereafter in the long term ( P < 0.01) to levels lower than baseline ( P = 0.023). Long-term PTH level was higher than baseline ( P = 0.012), with 11 patients showing PTH levels higher than normal (> 70 ng/L). Serum phosphorus level was increased in both the short term and the long term ( P < 0.01). Conclusions:Obese patients have 25-(OH)D deficiency/insufficiency before metabolic surgery and experience further decrease in the long term after surgery, despite a transient increase. Secondary increase in PTH level occurs in some of the patients after surgery. Long-term nutritional supplements and comprehensive nutritional management play important roles in postoperative management of obese patient.

OBJECTIVE: To investigate the effect of glycolytic enzyme pyruvate kinase type 2 (PKM2) on the proliferation and apoptosis of human leukemia HL-60 cells. METHODS: si-PKM2 plasmid was transfected into HL-60 cells (set as si-PKM2 group), and blank vector transfected cells were set as control group (si-Ctl group). The expression levels of PKM2 mRNA and protein in si-Ctl group and si-PKM2 group were detected by RT-qPCR and Western blot. CCK-8 cell detection kit was used to detect the proliferation ability of the cells in the two groups. Flow cytometry was used to detect the changes of cell cycle and apoptosis. Western blot and RT-qPCR were used to detect the changes of p-Akt and p-mTOR protein levels in PI3K/Akt/mTOR signaling pathway and the changes of glycolysis-related mRNA levels of the cells in the two groups. The changes in glucose consumption and lactic acid production of the cells were assayed. Over expressed PKM2, HL-60 cells were treated with PI3K inhibitor LY294002 or galactose, the changes in cell proliferation ability, cell cycle and apoptosis, as well as changes in glucose consumption and lactic acid production were detected. RESULTS: Interfered by si-PKM2, mRNA and protein levels of PKM2 in si-PKM2 group significantly decreased, and proliferation ability of the cells was also reduced (P<0.05). After PKM2 knockdown, the cells were significantly blocked at G CONCLUSION PKM2 knockdown can inhibit the proliferation and induce apoptosis of HL-60 cells, and its molecular mechanism may be related to the PKM2-mediated PI3K/Akt/mTOR-glycolysis, which suggesting that PKM2 may serve as a molecular target for the prevention and treatment of leukemia.
Apoptosis ; Cell Proliferation ; Glycolysis ; Humans ; Phosphatidylinositol 3-Kinases/metabolism* ; Pyruvate Kinase

Objective:To analyze the epidemiological characteristics of newly reported HIV/AIDS cases in Henan province during 2010-2018, and to provide theoretical evidence for strategies on prevention and control.Methods:Information about newly reported HIV/AIDS cases in Henan between January 1, 2010 and December 31, 2018 were collected from the National Comprehensive HIV/AIDS Information System. Excel 2010 and software SPSS 23.0 were used for data cleaning and statistical analysis.Results:During 2010-2018, a total of 41 223 HIV/AIDS cases were newly reported in Henan, and the number of newly reported HIV/AIDS cases increased from 3 087 in 2010 to 5 910 in 2018. Sexual contact was the predominant transmission route in newly reported HIV/AIDS cases, and the proportion of sexual transmission increased from 50.6 % (1 564/3 087) in 2010 to 98.8 % (5 839/5 910) in 2018. The proportion of heterosexual transmission firstly increased from 44.9 % (1 387/3 087) in 2010 and then declined to 65.7 % (3 885/5 910) in 2018, and the proportion of MSM behavior related transmission increased from 5.7 % (177/3 087) in 2010 to 33.1 % (1 954/5 910) in 2018. Among the newly reported heterosexual transmitted cases, the proportion of cases with non-marital heterosexual behaviors increased from 69.4 % (962/1 387) in 2010 to 91.7 % (3 562/3 885) in 2018 ( χ2=657.802, P<0.001). The number of newly reported HIV/AIDS cases in young people aged 15-24 years and old people aged ≥60 years increased year by year. The youth cases were mainly infected by MSM behavior transmission (52.7 %, 2 561/4 856), and heterosexual transmission was the main infection route of the elder cases (86.4 %, 5 907/6 833). Among the elder cases infected by heterosexual transmission, cases with non-marital heterosexual behaviors were mainly males, the number of male cases increased from 122 in 2010 to 738 in 2018, and sex composition ratio was between 82.2 % and 91.0 %, while the sex composition ratio of female cases was increased from 9.0 % (12/134) in 2010 to 17.8 % (160/898) in 2018. Conclusions:From 2010 to 2018, the number of newly reported HIV/AIDS cases in Henan increased. Sexual transmission had become the main transmission route. AIDS prevention and control should focus on the control of non-marital heterosexual transmission and MSM behavior transmission, and to pay more attention to young people and people aged 60 years and above.