Abstract: Objective　To explore the antiviral effect of baricitinib in the SARS-CoV-2 infection and influence on cytokine levels. Methods　Calu-3 cells were infected with SARS-CoV-2 at MOI of 0.1, and the levels of inflammatory cytokines (IL-6, IL-8, TNF-α and IL-1β), interferon β (IFN-β) and interferon-stimulated gene, IFIT2 in the infected cells were analyzed by qRT-PCR methods. At the same time, Calu-3 cells were infected with SARS-CoV-2 (MOI=0.1) after being treated with baricitinib for 2 hours. Cells were collected at 0, 24, 36, and 48 hours, and analyzed for the mRNA of the above genes in the drug-treated and untreated groups. Results　The mRNA levels of IL-6, TNF-a, IL-1β, IFN-β and IFIT2 in Calu-3 infected by SARS-CoV-2 cells were increased significantly. These cytokines were increased by nearly 100-fold post-infection 48 h compared with the control (P<0.000 1), and continued to increase with the infection time (P<0.001 or P<0.000 1). The increase of IL-8 mRNA level was not as significant as IL-6, TNF-α, IL-8, IL-1β, but it also showed a 2-4 folds increase. Baricitinib does not affect the level of viral RNA in Calu-3 cells after SARS-CoV-2 infection (P>0.05). However, baricitinib can significantly inhibit the up-regulation of IL-6 and TNF-α levels induced by SARS-CoV-2 infection (5.25-fold and 3.90-fold down-regulation, respectively, P<0.01), and has little effect on the levels of IL-8 and IL-1β . In addition, the drug could also significantly down-regulate the increase in IFN-β and IFIT2 levels caused by viral infection (10.51-fold and 90.78-fold down-regulation, respectively, P<0.000 1). Conclusions　Baricitinib inhibits the release of inflammatory cytokines to some extent, but it drastically down-regulates the expression of interferons and interferon-stimulated genes (ISGs), and has limited antiviral effect on SARS-CoV-2. Considering that interferon signal pathways play important roles on viral infection, caution should be exercised when using baricitinib to treat COVID-19 patients.

Objective: In this study, the feasibility of hot air-microwave combination drying method for processing Angelicae Sinensis Radix (ASR) was investigated and evaluated. Methods: Hot air temperature, drying time, microwave electric current, and microwave time were taken into consideration. The combined drying method was optimized based on the standards of ASR documented in Chinese Pharmacopoeia (2015 version). After that, microorganisms, heavy metals, harmful elements, and active ingredients (ferulic acid, coniferyl ferulate, senkyunolide I, senkyunolide H, senkyunolide A, E-ligustilide, Z-ligustilide, n-butylidenephthalide, riligustilide, and levistilide A) were detected to evaluate the quality of processed samples. Results: The optimal method was that samples were dried in hot air (70℃) for 20 h, then heated in microwave (100 mA) for 6 min. The number of microorganisms in processed samples equaled to sulfur-fumigated samples; Heavy metals and harmful elements were qualified after this processing; The new drying method for processing ASR had no significant effect on the active ingredient. Conclusion: After this process, the physical and chemical indicators of ASR are in line with the criteria of Chinese Pharmacopoeia. This hot air-microwave combination drying method should be a fast drying method for post-harvest handling of ASR.

OBJECTIVETo investigate dendritic cell (DC) malfunctions in patients with chronic hepatitis B (CHB) and try some means to restore the function in vitro.

METHODSTwelve CHB patients and 10 healthy people were enrolled in the study. Phenotype analysis and allogeneic mixed lymphocyte reaction (AMLR) assay of DC from these subjects were made. Enzyme-linked ELISpot method for detecting IFN-gamma-producing CD8 (+) T cells were used to evaluate the efficacy of DC loaded in vitro with HBsAg or HBcAg.

RESULTSDC from patients had a lower expression of co-stimulatory molecules and impaired AMLR capacity, but was restored partially by cytokine cocktail in vitro. Mature DC loaded with HBsAg or HBcAg showed a greater capacity for IFN-gamma-production than immature DC.

CONCLUSIONMalfunction of DC from CHB patients may be rescued by a cocktail of cytokines, and therapeutic DC vaccines loaded with HBV protein might be helpful to treat CHB patients.

Adult ; Case-Control Studies ; Cells, Cultured ; Cytokines ; immunology ; pharmacology ; Dendritic Cells ; drug effects ; immunology ; virology ; Female ; Hepatitis B Core Antigens ; immunology ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B virus ; immunology ; Hepatitis B, Chronic ; immunology ; virology ; Humans ; Male ; Viral Proteins ; immunology ; Young Adult

Chinese medicine pair (CMP) was frequently applied in traditional Chinese medicine (TCM) clinic, and its significance was shown in long-term clinical practices and many accumulated experiences. It is the unique combination of two relatively fixed Chinese medicines in TCM clinic with the basic feature and principle of TCM compatibility, is the most fundamental and the simplest form of TCM formulae with certain theory basis and combinatory reason, which is proven effective. And the unique combination is frequently used for achieving mutual reinforcement or detoxication. CMP is an intermediate point between single herb and many TCM formulae, reflecting the regularity of TCM formulae compatibility and connotation of differential treatment. This paper analyzed and summarized the basic characteristics, development process and research significance of CMP, which aims to lead the modern basic and applied research on compatibility theory of CMP.
Drug Interactions ; History, 16th Century ; History, 17th Century ; History, 18th Century ; History, 19th Century ; History, 20th Century ; History, 21st Century ; History, Ancient ; Medicine, Chinese Traditional ; history ; methods

OBJECTIVETo study the role of PCR technique in detection of mycobacterium tuberculosis in the samples from joint tuberculosis, and to evaluate the clinical value of PCR in diagnosis of joint tuberculosis.

METHODSFrom June 1993 to August 2001, PCR was used to detect DNA of mycobacterium tuberculosis, and the standard culture was applied to detect mycobacterium tuberculosis. Mycobacterium tuberculosis were respectively blindly by the two techniques in the samples obtained from 95 patients with joint tuberculosis (55 males and 40 females, the age ranging from 2 to 75 years, with an average of 34 years). The positive rate of mycobacterium tuberculosis detection was calculated.

RESULTSIn the detection of mycobacterium tuberculosis, positive rate was 82% (78/95) in PCR technique, and 16% (15/95) in standard culture technique. There were statistical differences between the two groups (chi2=67, P<0.001). The whole process of PCR amplification was automatic and could be finished within several hours, and the detecting time was considerably shorter.

CONCLUSIONPCR technique is a rapid, simple, sensitive and specific method for detection of mycobacterium tuberculosis in the samples of joint tuberculosis, showing more marked advantages than the standard culture technique. It is valuable in the early rapid diagnosis and differential diagnosis of joint tuberculosis.

Adolescent ; Aged ; Child ; Child, Preschool ; Culture Techniques ; methods ; Female ; Humans ; Male ; Middle Aged ; Mycobacterium tuberculosis ; genetics ; isolation & purification ; Polymerase Chain Reaction ; methods ; Tuberculosis, Osteoarticular ; diagnosis ; microbiology ; Young Adult

In order to evaluate the incidence of chromosomal abnormalities in patients with polycythemia vera (PV) accurately and to investigate the value of fluorescence in situ hybridization (FISH) technique in the detection of trisomies 8 and 9, conventional cytogenetics (CC) technique was used to detect karyotype and interphase FISH was used to detect trisomies 8 and 9 in 50 newly diagnosed PV and 8 normal individuals. The results showed that out of 50 cases, the 3 cases had chromosome karyotype abnormality, including trisomy 8, deletion of chromosome Y and inversion of chromosome 11 by CC technique. FISH method detected two cases of trisomy 8, including one case confirmed by CC technique, and one case of trisomy 9 neglected by CC technique. It is concluded that the incidence of chromosomal abnormalities in patients with PV is rare, and the incidence of trisomy 8 and trisomy 9 found in this study are relatively lower than that have been reported which may be related to the limited number of samples. Interphase FISH, as an important complement to CC, is a useful method for the detection of trisomies 8 and 9.
Adult ; Aged ; Chromosomes, Human, Pair 8 ; genetics ; Chromosomes, Human, Pair 9 ; genetics ; Cytogenetic Analysis ; Female ; Gene Deletion ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Polycythemia Vera ; genetics ; Trisomy

OBJECTIVETo investigate Siwu decoction and its composite drugs on the blood-deficiency model mice induced by acetylphenyhydrazine and cyclophosphamide.

METHODAcetylphenyhydrazine and cyclophosphamide were used to copy the blood-deficiency model mice. Automatic hematology analyzer was used to test the peripheral hemogram. Weighting method was used to test the liver index and spleen index; Kits for ATPase test was used to test the activities of Na+ - K+ - ATPase/Ca2+ - Mg2+ - ATPase in erythrocyte membrane. Flow cytometry was used to test the bone marrow cells' cell cycle.

RESULTAngelicar Sinensis Radix and Paeoniae Radix Alba had the most effective activity on the peripheral hemogram. Paeoniae Radix Alba, the drug pair including Angelicar Sinensis Radix and the drug- group including Paeoniae Radix Alba had the most effective activity on the liver index. All the drugs, drug-pairs, drug-groups and the formula had effect on the spleen index. To the activity of Na+ - K+ - ATPase in erythrocyte membrane, Rehmanniae Radix Praeparata, the drug-pairs and drug-groups including Angelicar Sinensis Radix exhibited the most effective activity. All the drugs, drug-pairs, drug-groups and the formula had the protective effect on the damaged bone marrow cells.

CONCLUSIONSiwu decoction and its composite drugs all had effect on the blood-deficiency model mice, but the action intensity was different. Angelicar Sinensis Radix and Paeoniae Radix Alba exhibited the most effective activity to the protection of the blood-deficiency model mice.

Adenosine Triphosphatases ; drug effects ; metabolism ; Angelica sinensis ; Animals ; Bone Marrow Cells ; drug effects ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cyclophosphamide ; Disease Models, Animal ; Drugs, Chinese Herbal ; metabolism ; pharmacology ; Female ; Glucosephosphate Dehydrogenase Deficiency ; chemically induced ; drug therapy ; metabolism ; pathology ; Liver ; drug effects ; metabolism ; Mice ; Mice, Inbred ICR ; Paeonia ; Phenylhydrazines ; Protective Agents ; pharmacology ; Spleen ; drug effects ; metabolism

OBJECTIVETo observe the difference between the combination therapy of alpha-interferon (IFN-alpha) therapy Yixuesheng Capsule and the monotherapy of IFN-alpha in treatment of chronic hepatitis B.

METHODA total of 288 patients with HBeAg-positive chronic hepatitis B proven by liver biopsy were included in this study. During the individualized therapy, they received hypodermic injection of IFN-alpha 1b, with 5 MU x time(-1) and three times x w(-1). Of them, 125 patients received combination therapy with Yixuesheng Capsule for three months, with 1.0 g/time and three times/d; and 163 patients received only IFN-alpha 1b (the IFN-alpha monotherapy group). After the course of therapy, all patients were followed up for at least 24 months. The intention-to-treat analysis was adopted for statistic analysis.

RESULTThe two groups showed no statistical significance by gender, age, liver necroinflammation grading, liver fibrosis staging, serum ALT levels, serum HBV DNA levels and IFN-alpha therapy course. The whole course and the 24-month follow-up visit cover all of 112 patients in the combination treatment group and 141 cases in the IFN-alpha monotherapy group. The response rates of the combination treatment group and the IFN-alpha monotherapy group were 48.0% (60/125) and 35.0% (57/163) (x = 4.980, P = 0.026) at the end of treatment, respectively, 45.6% (57/125) and 33.1% (54/163) (x2 = 4.645, P =0.031) at the end of 12-month-follow-up period, respectively, and 38.4% (48/125) and 32.5% (53/163) (x2 = 1.076, P = 0.300) at the end of 24-month follow-up period, respectively.

CONCLUSIONThe combination treatment with IFN-alpha and Yixuesheng Capsule shows a slightly better sustained efficacy on HBeAg-positive chronic hepatitis B patients compared with IFN-alpha monotherapy.

Adult ; Capsules ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Follow-Up Studies ; Hepatitis B, Chronic ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Treatment Outcome

To evaluate the frequency of jak2v617f mutation and analyze its correlation with clinical features of primary thrombocythemia (PT) patients, the mutation was detected by allele specific polymerase chain reaction (AS-PCR), the clinical and laboratory data in 66 PT patients with and without jak2v617f mutation were compared and clinical features of these PT patients were retrospectively analyzed. The results showed that among 66 patients, 27 patients (41%) had the mutation. The patients carrying the mutation displayed higher bone marrow erythropoiesis (26.9%+/-9.4% vs 16.3%+/-8.7%, p<0.05), higher ratio of granulopoiesis/erythropoiesis (2.9+/-1.8 vs 5.2+/-2.9, p<0.05) and higher incidence of microvascular disturbances (29.6% vs 5.1%, p<0.05). However, the age, gender, leukocyte and platelet counts, hemoglobin level, bone granulopoiesis, splenomegaly, history of thrombosis and hemorrhage had no difference between patients with and without the mutation. It is concluded that the frequency of jak2v617f mutation in primary thrombocythemia patients is 41%. The presence of the jak2v617f mutation is associated with a higher bone marrow erythropoiesis in primary thrombocythemia patients at diagnosis.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Alleles ; Erythropoiesis ; Female ; Humans ; Janus Kinase 2 ; genetics ; Male ; Middle Aged ; Point Mutation ; Polymerase Chain Reaction ; methods ; Retrospective Studies ; Thrombocythemia, Essential ; diagnosis ; genetics ; Young Adult

OBJECTIVETo explore the association between HBV genotype and chronic/severe liver disease with HBV infection in Chinese patients.

METHODSSerum samples were collected from 2922 patients with HBV infection. HBV genotyping was performed with type-specific primers polymerase chain reaction, and the virological and biochemical markers were detected, which differences in the genotypes between various clinical types of HBV infection and liver function and virological markers between various HBV genotyping were analyzed.

RESULTSThe genotype B, C, BC combinations, D of 2922 patients with HBV infection accounted for 15.9%, 83.5%, 0.41%, 0.21% respectively. The ratio of genotype B in acute hepatitis group was higher (P = 0.003), which the ratio of genotype C in the cirrhosis group and the hepatocellular carcinoma group was higher (P = 0.000, 0.000). The difference in ratio of genotype C was not statistically significant between acute-on-chronic liver failure group and chronic hepatitis group. HBeAg-positive rate, viral load and liver function markers of B, C genotype group in acute hepatitis group and chronic hepatitis group were not significant different. HBeAg-positive rates of genotype C in acute-on-chronic liver failure group, cirrhosis group, hepatocellular carcinoma group were higher than that of genotype B (P = 0.000, 0.024, 0.003). Viral load of genotype C in hepatocellular carcinoma group was higher than that of genotype B (P = 0.025). Cholinesterase levels of genotype C in the acute-on-chronic liver failure group and the hepatocellular carcinoma group was lower than that of genotype B (P = 0.0004, 0.02).

CONCLUSIONThere were HBV genotype B, C, B/C combinations and D in Chinese patients with HBV infection, with genotype B and C being the major ones. Compared with HBV genotype B, genotype C in Chinese patients with HBV infection was more likely to chronic infection, evolved to cirrhosis and hepatocellular carcinoma, but genotype difference was not observed in occurrence of acute-on-chronic liver failure. Genotype was not significant effect in acute and chronic hepatitis B, but HBeAg-positive rate/viral load was higher and liver damage was more severe in severe and end-stage genotype C HBV infection patients.

Adult ; Age Factors ; Animals ; Asian Continental Ancestry Group ; genetics ; Cricetinae ; End Stage Liver Disease ; genetics ; Female ; Genotype ; Hepatitis B ; virology ; Hepatitis B e Antigens ; Hepatitis B virus ; Hepatitis B, Chronic ; virology ; Humans ; Liver Cirrhosis ; genetics ; Liver Diseases ; genetics ; Liver Neoplasms ; genetics ; Male ; Polymerase Chain Reaction ; Sex Factors