Modified Xiaoyaosan Alleviates Depression-like Behaviors by Regulating Activation of Hippocampal Microglia Cells in Rat Model of Juvenile Depression / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism of Baihuan Xiaoyao Decoction （Xiaoyaosan added with Lilii Bulbus and Albiziae Cortex） in alleviating depression-like behaviors of juvenile rats by regulating the polarization of microglia. MethodSixty juvenile SD rats were randomized into normal control， model， fluoxetine， and low-， medium-， and high-dose （5.36， 10.71， 21.42 g·kg-1， respectively） Baihuan Xiaoyao decoction groups. The rat model of juvenile depression was established by chronic unpredictable mild stress （CUMS）. The sucrose preference test （SPT） was carried out to examine the sucrose preference of rats. Forced swimming test （FST） was carried out to measure the immobility time of rats. The open field test （OFT） was conducted to measure the total distance， the central distance， the number of horizontal crossings， and the frequency of rearing. Morris water maze （MWM） was used to measure the escape latency and the number of crossing the platform. The immunofluorescence assay was employed to detect the expression of inducible nitric oxide synthase （iNOS， the polarization marker of M1 microglia） and CD206 （the polarization marker of M2 microglia）. Real-time polymerase chain reaction was employed to determine the mRNA levels of iNOS， CD206， pro-inflammatory cytokines ［tumor necrosis factor （TNF）-α， interleukin （IL）-1β， and IL-6］ and anti-inflammatory cytokines （IL-4 and IL-10） in the hippocampus. Western blotting was employed to determine the protein levels of iNOS and CD206 in the hippocampus. The levels of IL-4 and IL-6 in the hippocampus were detected by enzyme-linked immunosorbent assay. ResultCompared with the normal control group， the model rats showed a reduction in sucrose preference （P<0.05）， an increase in immobility time （P<0.05）， decreased motor and exploratory behaviors （P<0.05）， and weakened learning and spatial memory （P<0.05）. In addition， the model rats showed up-regulated mRNA and protein levels of iNOS and mRNA levels of IL-1β， IL-6， and TNF-α （P<0.05）. Compared with the model group， Baihuan Xiaoyao decoction increased the sucrose preference value （P<0.05）， shortened the immobility time （P<0.01）， increased the motor and exploratory behaviors （P<0.05）， and improved the learning and spatial memory （P<0.01）. Furthermore， the decoction down-regulated the positive expression and protein level of iNOS， lowered the levels of TNF-α， IL-1β， and IL-6 （P<0.01）， promoted the positive expression of CD206， and elevated the levels of IL-4 and IL-10 （P<0.01） in the hippocampus of the high dose group. Moreover， the high-dose Baihuan Xiaoyao decoction group had higher sucrose preference value （P<0.01）， shorter immobility time （P<0.01）， longer central distance （P<0.01）， stronger learning and spatial memory （P<0.01）， higher positive expression and protein level of iNOS （P<0.01）， lower levels of TNF-α， IL-1β， and IL-6 （P<0.05， P<0.01）， lower positive expression and mRNA level of iNOS （P<0.05）， and higher levels of IL-4 and IL-10 （P<0.05， P<0.01） than the fluoxetine group. ConclusionBaihuan Xiaoyao decoction can improve the depression-like behavior of juvenile rats by inhibiting the M1 polarization and promoting the M2 polarization of microglia in the hippocampus.

Rapid detection of zearalenone in Coicis Semen based on ELISA / 中国中药杂志

Zearalenone(ZEN) is a toxic metabolite produced by Fusarium culmorum, F. graminearum, F. tricinctum, and other fungi, with estrogenic characteristics. Exposure to or ingestion of ZEN during pregnancy can cause reproductive dysfunction, miscarriage, stillbirth, and malformation, and seriously endanger human life and health. The detection methods for ZEN in the Chinese Pharmacopoeia(2020 edition) are liquid chromatography(LC) and liquid chromatography-mass spectrometry(LC-MS), and it is stipulated that ZEN should not exceed 500 μg in 1 000 g of Coicis Semen. Although these detection methods by instruments can achieve the qualitative and quantitative analysis of ZEN in Coicis Semen, their high detection cost and long periods hinder the rapid screening of a large number of samples in the field. In this study, the synthesized ZEN hapten was conjugated with bovine serum albumin(BSA) and ovalbumin(OVA) to obtain the complete ZEN antigen. By virtue of antibody preparation techniques, ZEN monoclonal antibody 4F6 was prepared, which showed 177.5%, 137.1%, and 109.7% cross-reactivity with ZEN structural analogs zearalanol, zearalenone, and α-zearalenol, respectively, and no cross-reactivity with other fungal toxins such as aflatoxin. Direct competitive enzyme-linked immunosorbent assay(dcELISA) based on ZEN monoclonal antibody 4F6 was developed for the determination of ZEN in Coicis Semen with an IC_(50) of 1.3 μg·L~(-1) and a detection range of 0.22-21.92 μg·L~(-1). The recoveries were 83.91%-105.3% and the RSD was 4.4%-8.0%. The established dcELISA method was used to determine the ZEN residuals in nine batches of Coicis Semen samples, and the results were validated by LC-MS. The correlation between the two detection methods was found to be 0.993 9, indicating that the established dcELISA could be used for the rapid qualitative and quantitative detection of ZEN residuals in Coicis Semen.

Clinical features of six patients with Neurodevelopmental disorder with spastic diplegia and visual defects caused by CTNNB1 gene mutation / 中华检验医学杂志

Objective:To investigate the clinical features and characteristics of gene mutation of patients with neurodevelopmental disorder caused by CTNNB1 gene. Method:Genetic mutation analysis of the patients were obtained by using the whole exome sequencing and Sanger sequencing. We reviewed the literatures for the clinical and genetic features of CTNNB1 related neurodevelopmental disorder. Results:Six inpatients, three boys and three girls, who came for speech impairment motor delay were included in this study. The average age for the patients was 17.8±11.1 months. The main clinical manifestations of the patients were craniofacial dysmorphism, microcephaly, hypertonia or spasm, speech impairment motor delay, esotropia and valgus. WES showed that 6 patients carried de novo mutations of CTNNB1 gene, which were c.1057delA, c.1493_1494insA, c.418_424del, c.1985_1988del, c.1420C>T and c.1550T>C. No abnormality was found in the patients′ parents. Conclusions:The clinical manifestation of CTNNB1 related neurodevelopmental disorder involves multiple systems. We found five unreported variants and expanded the variation spectrum of the CTNNB1 gene.

The mechanism of proprotein convertase subtilisin/kexin type 9 in the inflammatory response induced by Helicobacter pylori infection / 中华消化杂志

Objective:To explore the mechanism of proprotein convertase subtilisin/kexin type 9 (PCSK9) in the inflammatory response induced by Helicobacter pylori ( H. pylori) infection. Methods:From May 1, 2020 to January 31, 2021, 60 patients with gastritis (30 H. pylori positive and 30 H. pylori negative)and 30 healthy individuals, who initially visited the Department of Gastroenterology, Shiyan Taihe Hospital were collected, and their serum PCSK9 levels were detected. Normal gastric epithelial cell line GES-1 and macrophages induced from THP-1 cells, and GES-1 infected with H. pylori were selected to prepare different supernatant media. Phosphate buffer saline empty medium (negative control group), normal GES-1 cell supernatant medium ( H. pylori-uninfected GES-1 group), H. pylori infected GES-1 cell supernatant medium ( H. pylori infected GES-1 group), H. pylori infected GES-1 cell supernatant + PCSK9 neutralizing antibody medium (anti PCSK9 group), H. pylori infected GES-1 cell supernatant+ human immunoglobulin G medium (isotype control group) were established. The differences between H. pylori infected GES-1 group and H. pylori-uninfected GES-1 group, negative control group, anti PCSK9 group and isotype control group in number of migrated macrophages, relative expression level of CC chemokine receptor ( CCR2), the levels of released interleukin(IL)-6 and cell necrosis factor (TNF)- α, level of CD8 + T cell membrane phosphorylation, and the number of macrophage colonies were determined by Transwell assay, real time fluorescence quantitative polymerase chain reaction, plate colony assay, H. pylori and phagocytosis lysosome co-localization assay. The regulating mechanism of PCSK9 in H. pylori infection induced inflammation was analyzed. Independent sample t test was used for statistical analysis. Results:The serum level of PCSK9 of patients with H. pylori positive gastritis was higher than that of patients with H. pylori negative gastritis and healthy individuals ((384.00±57.57) g/L vs. (208.80±48.89) and (176.10±47.14) g/L), and the differences were statistically significant ( t=12.71 and 15.31; both P<0.001). Compared with negative control group, H. pylori standard strain and 4 isolated H. pylori strains could stimulate GES-1 to secrete PCSK9 ((1 267.00±287.50) g/L vs.(2 717.00±199.20), (4 858.00±302.40), (3 167.00±334.20), (6 075.00±597.30), (4 283.00±331.20) g/L), and the differences were statistically significant( t=10.15, 21.09, 10.56, 17.77, 16.85, all P<0.001). The number of migrated macrophages, CCR2 mRNA expression level in macrophage, expression levels of IL-6 and TNF-α, and the number of macrophage colonies of H. pylori-infected GES-1 group were all higher than those of H. pylori-uninfected GES-1 group and negative control group (132.20±5.67 vs.84.83±4.62, 39.83±4.12; 8.66±0.94 vs. 6.52±0.47 and 1.00±0.09, (281.00±8.56) ng/L vs. (115.00±7.72) and (64.00±5.44) ng/L, (619.80±18.47) ng/L vs.(373.30±12.85)and (225.70±6.44) ng/L, (357.00±16.31) colony forming unit (CFU) vs. (134.80±8.64) and (74.17±9.68) CFU), and the differences were statistically significant ( t=15.85, 32.27; 4.96, 19.79; 35.28, 52.43; 26.84, 49.37; 29.49, 36.53; all P<0.001). The percentage of co-localization of H. pylori and phagocytosis lysosome, and the expression of cell membrane CD3ζ Tyr142, granzyme B and perforin in CD8 + T cell of H. pylori-infected GES-1 group were lower than that of H. pylori-uninfected GES-1 group ((15.33±1.86)% vs. (34.50±3.72)% and (65.67±3.56)%, 464.20±120.80 vs. 1 924.00±262.10 and 2 390.00±484.10; (6.41±0.42)% vs.(17.37±0.73)% and (26.60±1.57)%; (6.84±1.37)% vs.(14.53±0.48)% and (26.22±1.21)%), and the differences were statistically significant( t=11.27 and 30.70, 12.39 and 9.45, 30.50 and 31.90, 25.96 and 13.00; all P<0.001). The number of migrated macrophages, the relative expression level of CCR2 mRNA, the expression levels of IL-6 and TNF-α, and the number of macrophage colonies of anti-PCSK9 group were all lower than those of isotype control group (72.50±4.97 vs. 128.30±6.74, 0.82±0.06 vs. 1.00±0.08, (85.50±4.37) ng/L vs. (277.70±8.98) ng/L, (291.80±13.69) ng/L vs. (615.30±12.65) ng/L, (111.50±10.21) CFU vs. (346.20±18.04) CFU), and the differences were statistically significant ( t=16.33, 4.40, 47.13, 42.50 and 27.73, all P<0.001). The percentage of co-localization of H. pylori and phagocytosis lysosome, the expression levels of CD3ζ Tyr142, granzyme B and perforin of anti-PCSK9 group were all higher than those of isotype control group ((51.05±3.03)% vs. (16.71±1.91)%, 2 948.00±384.00 vs. 1 156.00±178.60, (53.88±3.86)% vs. (5.88±0.93)%, (32.80±2.07)% vs. (6.83±0.54)%), and the differences were statistically significant ( t=23.49, 10.36, 29.60 and 29.76, all P<0.001). Conclusions:H. pylori can inhibit CD8 + T activation and cytotoxicity by inducing the release of PCSK9 from gastric epithelial cells, and can also recruit macrophages, activate nuclear factor-κB signal axis to up-regulate the level of released inflammatory factors from macrophages, inhibit the phagocytosis and killing effects of macrophages, so as to regulate the inflammatory response.

Clinical and genetic features of a patient with COG5-CDG caused by COG5 gene variation / 中华检验医学杂志

The patient in this study was a 3 years 8 month old boy. The patient presented with facial dysmorphism including wide eye distance and flat nose. The major clinical manifestations were poor response, backward language and motor development; and his fingers cannot be bent. Moreover, the patient′s hands were also uncoordinated. In addition the patient suffered from congenital myopia and nystagmus; and the teeth were fall off easily. The abnormal reproductive system was characterized by small penis and small testicle. No obvious abnormality was found in liver and kidney function and serum immunoglobulin level through laboratory biochemical test. The results of the spine X-Ray examination indicates scoliosis. Results from brain MRI showed cerebellar dysplasia. Compound heterozygous variants in COG5 gene (c.1039C>T and c.928+3A>G), each inheranted from his parents were found in this patient by high-throughput sequencing and Sanger sequencing. After a clear diagnosis, the patient received language rehabilitation training and motor rehabilitation training. In this study, we found two new variants in COG5 gene and increased the mutation spectrum of this gene.

Detection and analysis of hepatitis B virus serum markers in children treated in Wuhan Children's Hospital / 公共卫生与预防医学

Objective To investigate the status of hepatitis B virus (HBV) infection in children in Wuhan, and to analyze the expression pattern and distribution of serum markers. Methods Five serum markers of HbsA, HbsAb, HbeAg, HbeAb and HBcAb were detected by electrochemiluminescence immunoassay in 67 027 children aged 0-18 years including inpatients, outpatients, and physical examinees in Wuhan Children's Hospital. SPSS24.0 statistical software was used to analyze the results by age and gender. Results The “all negative” detection rate of all 67,027 children was 18.98%. There was a significant difference in the positive rate of HBcAb between male and female. The positive rate of HBcAb was higher in 0~28 days and 1~12 months group and decreased significantly after 1 year old. The positive rate of HBcAb was 5.02% in 1-14 years old but increased slightly in 15-18 years old. Among HBsAb positive children, the positive rate of HBsAb reached the peak of 95.65% in 1~2 years old group and the lowest of 68.90% in 6~14 years old group, and gradually decreased before 15 years old. Among the children with HBsAb concentration ≥100 IU/L, the proportion of 1~2 years old group was the highest (76.99%), and the proportion of 6~14 years old group was the lowest (40.99%). A total of 20 HBsAb serum marker expression patterns were detected, and the detection rates of “single HBsAb+”, “all negative”, “HBsAb+/HBcAb+”, and “HBsAb+/HBeAb+/HBcAb+” were 71.40%, 18.98%, 4.80% and 4.20%, respectively. Among them, 11 kinds of uncommon expression patterns were detected, and 9 kinds of uncommon expression patterns were detected in neonates, with a detection rate of 1.21%, which was higher than that in other age groups. Among all serological patterns, only the detection rate of “single HBcAb+” showed a statistical difference between male and female. Conclusion The HBV infection rate in all ages of 0~18 years old children in Wuhan is low. “Single HBsAb+” is the main serological pattern, and the concentration distribution of HBsAb is mostly in the range of 100-999 IU /L. There is a high “all negative” detection rate. School-age children should be inoculated with hepatitis B vaccine, which may be beneficial to reduce the risk of infection.

Crotalaria ferruginea extract attenuates lipopolysaccharide-induced acute lung injury in mice by inhibiting MAPK/NF-κB signaling pathways

Objective: To evaluate the anti-inflammatory activity of Crotalaria ferruginea extract (CFE) and its mechanism. Methods: An intratracheal lipopolysaccharide (LPS) instillation-induced acute lung injury (ALI) model was used to study the anti-inflammatory activity of CFE in vivo. The LPS-induced shock model was used to analyze the effect of CFE on survival. LPS-stimulated RAW264.7 cell model was used to investigate the anti-inflammatory activity of CFE in vitro and the effects on mitogen-Activated protein kinase (MAPK) or nuclear factor-κB (NF-κB) signaling pathways. Results: CFE administration decreased the number of inflammatory cells, reduced the levels of tumor necrosis factor-α (TNF-A), monocyte chemotactic protein-1 (MCP-1), interleukin-6 (IL-6), and interferon-γ, and diminished protein content in the bronchoalveolar lavage fluid of mice. CFE also reduced lung wet-To-dry weight ratio, myeloperoxidase, and lung tissue pathological injury. CFE pre-Administration improved the survival rate of mice challenged with a lethal dose of LPS. CFE reduced LPS-Activated RAW264.7 cells to produce nitric oxide, TNF-α, MCP-1, and IL-6. Furthermore, CFE inhibited nuclear translocation and phosphorylation of NF-κB P65, extracellular signal-regulated kinase, c-Jun N-Terminal kinases, and P38 MAPKs. Conclusions: CFE exhibits potent anti-inflammatory activity in LPS-induced ALI mice, LPS-shock mice, and RAW264.7 cells, and its mechanism may be associated with the inhibition of NF-κB and MAPK signaling pathways. Crotalaria ferruginea may be a useful therapeutic drug for the treatment of ALI and other respiratory inflammations.

Crotalaria ferruginea extract attenuates lipopolysaccharide-induced acute lung injury in mice by inhibiting MAPK/NF-κB signaling pathways

Objective: To evaluate the anti-inflammatory activity of Crotalaria ferruginea extract (CFE) and its mechanism. Methods: An intratracheal lipopolysaccharide (LPS) instillation-induced acute lung injury (ALI) model was used to study the anti-inflammatory activity of CFE in vivo. The LPS-induced shock model was used to analyze the effect of CFE on survival. LPS-stimulated RAW264.7 cell model was used to investigate the anti-inflammatory activity of CFE in vitro and the effects on mitogen-Activated protein kinase (MAPK) or nuclear factor-κB (NF-κB) signaling pathways. Results: CFE administration decreased the number of inflammatory cells, reduced the levels of tumor necrosis factor-α (TNF-A), monocyte chemotactic protein-1 (MCP-1), interleukin-6 (IL-6), and interferon-γ, and diminished protein content in the bronchoalveolar lavage fluid of mice. CFE also reduced lung wet-To-dry weight ratio, myeloperoxidase, and lung tissue pathological injury. CFE pre-Administration improved the survival rate of mice challenged with a lethal dose of LPS. CFE reduced LPS-Activated RAW264.7 cells to produce nitric oxide, TNF-α, MCP-1, and IL-6. Furthermore, CFE inhibited nuclear translocation and phosphorylation of NF-κB P65, extracellular signal-regulated kinase, c-Jun N-Terminal kinases, and P38 MAPKs. Conclusions: CFE exhibits potent anti-inflammatory activity in LPS-induced ALI mice, LPS-shock mice, and RAW264.7 cells, and its mechanism may be associated with the inhibition of NF-κB and MAPK signaling pathways. Crotalaria ferruginea may be a useful therapeutic drug for the treatment of ALI and other respiratory inflammations.

Evaluation of the performance of systems for whole blood C-reactive protein detection: a multi-center study / 中华检验医学杂志

Objective:To explore the performance of the commonly used whole blood C-reactive protein (CRP) detection systems and give related recommendation on the performance requirements of detection systems.Methods:A total of 7 540 venous blood samples from 26 maternal, child and children′s hospitals were collected to conduct this multi-center study on the analytical performance of 5 commonly used whole blood CRP detection systems from March to April in 2019. The blank check, carryover, repeatability, intermediate precision, linearity, sample stability, influence of hematocrit/triglyceride/bilirubin, comparison with SIEMENS specific protein analyzer and trueness were evaluated. The 5 systems included BC-5390CRP autohematology analyzer, AstepPLUS specific protein analyzer, Ottoman-1000 Automated Specific Protein POCT Workstation, i-CHROMA Immunofluorometer equipment Reader and Orion QuikRead go detecting instrument. The 5 systems were labeled as a, b, c, d and e randomly.Results:Within the 5 systems, all values of blank check were less than 1.00 mg/L, the carryovers were lower than 1.00%. The repeatability of different ranges of CRP concentrations including 3.00-10.00, 10.00-30.00 and>30.00 mg/L were less than 10.00%, 6.00% and 5.00%, respectively, and the intermediate precision was less than 10.00%. The linearity correlation coefficients of the 5 systems were all above 0.975, while the slope was within 0.950-1.050. Whole blood samples were stable within 72 hours both at room temperature (18-25 ℃) and refrigerated temperature (2-8 ℃). The CRP results were rarely influenced by high triglyceride or bilirubin, except for the immmunoturbidimetric test based on microparticles coated with anti-human CRP F(ab) 2 fragments. When triglyceride was less than 15.46 mmol/L, the deviation of CRP was less than 10.00%. When bilirubin was less than 345.47 μmol/L, the deviation of CRP was less than 10.00%. CRP was more susceptible to Hct on the systems without Hct correction. The deviation of CRP between different Hct dilution concentration and 40% dilution concentration can reach as high as 67.48%. The correlation coefficients ( r) of 5 systems were all more than 0.975 in the range of 0-300.00 mg/L compared with Siemens specific protein analyzer. All systems passed the trueness verification using the samples with specified values of 12.89 and 30.60 mg/L. Conclusion:The performance of 5 systems can basically meet the clinical needs, but it is suggested that the whole blood CRP detection system without automatic Hct correction should be modified manually.

Direct interaction between Urease B of Helicobacter pylori and TLR2 negatively regulates immune fucntions of macrophages / 中华微生物学和免疫学杂志

Objective:To evaluate the regulatory role and potential mechanism of Urease B(UreB) on macrophages.Methods:Bone marrow-derived macrophages (M0) were stimulated by recombinant UreB protein and then flowcytometry and ELISA were used to detect the apoptosis, polarization and antigen presentation-related biomarkers expression. CD4 + T cell co-culture assay, CFSE stain and flowcytometry were used to evaluate the impacts of UreB on antigen presentation capacity of macrophages. Truncated UreB protein, NanoBiT assay and co-immunoprecipitation were used to identify the binding sites of UreB to TLR2. Results:UreB promoted apoptosis and skewed macrophages from M1 to M2 in the presence of M1-inducer LPS. Moreover, UreB inhibited the expression of antigen presentation biomarkers, MHCⅡ and CD86 on macrophages, and further inhibited the proliferation and IFN-γ expression of CD4 + T cells. Molecular analyses revealed that the binding between seven carboxy-terminal amino acid residues of UreB and TLR2 were required for the UreB-mediated inhibitory effects. Conclusions:The findings in this study demonstrate that UreB mainly depends on the binding between seven carboxy-terminal amino acid residues and TLR2 to perform immune-suppressive activities, and which may provide valuable information for the design and optimization of UreB-based vaccines against Helicobacter pylori infection.

The study on proinflammatory effect of Mycobacterium tuberculosis PE_PGRS60 protein through JNK-COX2 axis / 中华微生物学和免疫学杂志

Objective:To investigate the mechanism of PE_PGRS60 protein in the pathogenesis of Mycobacterium tuberculosis infection. Methods:The cloned and purified PE_PGRS60 protein from Mycobacterium tuberculosis was used to stimulate RAW264.7 cells. The expression of cyclooxygenase 2(COX2) mRNA and protein was detected by qRT-PCR and Western blot, respectively. The signal pathways that may regulate the expression of COX2 were screened, and the expression of inflammatory cytokines induced by PE_PGRS60 was detected by ELISA. The level of cell death was measured by lactate dehydrogenase(LDH) release test and flow cytometry PI staining. Western blot was used to detect the expression of COX2 in Peripheral blood mononuclear cell(PBMC) from active tuberculosis patients. Results:PE_PGRS60 protein was found to promote the expression of COX2 in RAW264.7 cells and activate the three major members of the mitogen-activated protein kinase(MAPK) family: extracellular regulated protein kinase(ERK), p38 and c-Jun N-terminal kinase(JNK). Interestingly, only JNK-IN-7, the inhibitor of JNK was observed to suppress the up-regulation expression of COX2 induced by PE_PGRS60. This up-regulated expression of COX2 was also found in PBMCs from active tuberculosis patients. The COX2 inhibitor celecoxib can effectively block the expression of the inflammatory factors IL-1β, TNF-α and IL-6 induced by PE_PGRS60 and promote macrophage death.Conclusions:PE_PGRS60 can promote macrophages to release inflammatory factors by activating JNK/COX2 signal axis. Some macrophages still die under the protection of COX2.

Effects of breast feeding on serum ghrelin levels and insulin sensitivity of small for gestational age infants / 中华实用儿科临床杂志

Objective:To investigate the changes of plasma ghrelin levels and insulin(INS) sensitivity of full-term infants small for gestational age (SGA) and the effects of breast feeding on it.Methods:Full-term SGA hospitalised in the Department of Neonatology, Wuhan Children′s Hospital from October 2014 to April 2019 were re-cruited as the SGA group (120 cases), with full-term infants appropriate for gestational age (AGA) born in the same period as the AGA group (96 cases) in this study with recorded birth weight and length.The levels of fasting blood glucose (FG), triglyceride (TG), low density lipoprotein (LDL), high density lipoprotein (HDL), INS and ghrelin were measured 7 days after birth.Homeostasis model assessment-insulin resistance (HOMA-IR) was calculated.The SGA group was subdivided into breast feeding group and formula feeding group.The above indexes were tracked and mea-sured in the 3 rd and 6 th month, respectively, and their growth parameters were recorded. Results:There were no diffe-rences in serum FG, TG, LDL and HDL levels between the SGA and the AGA group (all P>0.05). Compared with the AGA group, the serum INS[(4.21±0.83) mIU/L vs.(3.54±1.10) mIU/L], ghrelin levels[(0.80±0.23) μg/L vs.(0.69±0.19) μg/L] and HOMA-IR (0.85±0.25 vs.0.72±0.25) increased in the SGA group, the differences were statistically significant (all P<0.05). Serum INS, HOMA-IR and ghrelin levels changed with the duration of breast feeding, the differences were statistically significant( F=12.394, 9.810, 5.531, all P<0.05). Conclusions:The serum ghrelin levels of SGA infants increased and INS sensitivity decreased.Breastfeeding can decrease levels of serum INS, HOMA-IR and ghrelin, and can improve INS sensitivity of SGA infants.

Safety of Thread Embedding Acupuncture Therapy: A Systematic Review / 中国结合医学杂志

OBJECTIVE@#To evaluate the safety of thread embedding acupuncture therapy (TEAT) and discuss the prevention and treatment of some adverse events (AEs).@*METHODS@#Review of databases, including China National Knowledge Infrastructure (CNKI), CBMdisc, Wanfang, VIP databases and English literature published in PubMed, MEDLINE, EMBASE and Web of Science, were searched from their inception to January 2020, randomized controlled trials (RCTs) and case reports in which AEs with TEAT were included. Cochrane Collaboration's tool and RevMan V.5.3.3 software were used to evaluate the quality of the studies.@*RESULTS@#A total of 61 studies (45 RCTs and 16 case reports) with 620 cases of AEs were included in this review. These studies were published in two countries: China and South Korea. Twenty eight kinds of AEs were summarized. The most common AEs were induration, bleeding and ecchymosis, redness and swelling, fever, and pain. They were accounted for 75.35% (425/564) in the review, and most of them were mild. The rarest AEs were epilepsy, irregular menstruation, skin ulcer, thread malabsorption, and fat liquefaction, with 1 case each. But not all of them had clear causal relationship with TEAT. Most of the AEs were local reactions [with incidence of 9.83% (480/4,882)] and systemic reactions accounted for only 1.27% (62/4,882). Although the included studies showed that AEs were very commonly encountered (11.09%), only 5 cases of severe AEs reported from 2013 to 2017 (0.1%) by using catgut thread, which are rarely seen nowdays with the wide use of new absorbable surgical suture. All of the severe AEs were recovered after symptomatic treatment with no sequelae.@*CONCLUSIONS@#The evidence showed that TEAT is a relatively safe and convenient therapy especially since application of new absorbable surgical suture. Improving practitioner skills, regulating operations, and paying attention to the patients' conditions may reduce the incidence of AEs and improve safety of TEAT.

Factors Related to Curative Effect of Brain-computer Interface Training on Upper Limb Paralysis after Stroke / 中国康复理论与实践

Objective:To explore the factors affecting curative effect of motor imagery brain-computer interface (MI-BCI) training on upper limb paralysis for subacute stroke patients. Methods:From January, 2018 to July, 2019, 23 inpatients with post-stroke upper limb paralysis accepting MI-BCI training were reviewed. The gender, age, course of disease, aphasia, location and nature of lesion, history of Botulinum toxin, hemisphere injured and modified Ashworth Scale (MAS) score of affected fingers were recorded, and they were assessed with Fugl-Meyer Assessment-Upper Extremities (FMA-UE) before and four weeks after MI-BCI training. According to improvement of FMA-UE wrist and hand scores (≥ 2), the patients were divided into effective group (n = 11) and inefficacy group (n = 12). Results:The MAS scores before MI-BCI training (t = 2.677, P < 0.05) and history of botulinum toxin (Z = 0.000, P < 0.05) were more in the inefficacy group than in the efficacy group. FMA-UE scores (total and dimensions) after training were correlated to their baseline levels (r > 0.831, P < 0.01), FMA-UE total scores (Eta = 0.453, P < 0.05) and upper arms scores (Eta = 0.506, P < 0.05) were correlated to aphasia, FMA-UE scores of hands were correlated with MAS (r = -0.521, P < 0.05). Conclusion:Poor baseline motor function, spasticity and complication with aphasia were the factors unfavorable to MI-BCI training for subacute stroke patients with upper limb paralysis.

High-risk human papillomavirus 16 E6 protein regulates tumor-associated macrophages through exosomal heat shock protein 70 to promote cervical cancer cell C33A proliferation, invasion and migration / 中华微生物学和免疫学杂志

Objective:To investigate the possible molecular mechanism of high-risk human papillomavirus (HPV) 16 E6 protein in promoting the proliferation, invasion and migration of cervical cancer cells.Methods:C33A cells were infected with HPV E6 protein overexpression lentivirus to construct a stable C33A-E6 cell line. The expression of heat shock protein 70 (HSP70) was detected by qRT-PCR and Western blot. Degradation rates of HSP70 at mRNA and protein levels were detected by qRT-PCR and Western blot at different time points after actinomycin D (ActD) and actinomycin ketone (CHX) treatment. Exosomes were extracted by hypervelocity centrifugation and the concentrations of HSP70 in exosomes were detected by enzyme-linked immunosorbent assay (ELISA). The concentrations of TNF-α, IL-1β, IL-6 and IL-10 in the supernatants of THP-1 cell culture that treated with exosomes isolated from C33A and C33A-E6 cells after interfering HSP70 expression were detected by ELISA. C33A cells were co-cultured with exosome-treated THP-1 cells to evaluate the changes in cell proliferation with CCK-8 method. The invasion and migration of C33A cells were assessed by Transwell assay.Results:Compared with the C33A cells, the expression of HSP70 at mRNA level in C33A-E6 cells was significantly increased ( P<0.05), but no significant change was observed at the protein level ( P>0.05). No significant difference in the degradation rate of HSP70 at mRNA or protein level was detected between C33A and C33A-E6 groups after treating with ActD and CHX for different times, but the concentration of HSP70 in exosomes was significantly increased in the C33A-E6 group ( P<0.001). The exosomes secreted by C33A-E6 cells could enhance the secretion of IL-6, TNF-α and IL-10 by THP-1 cells ( P<0.001), and the THP-1 cells treated with the exosomes could promote the proliferation, migration and invasion of C33A cells ( P<0.001). However, interfering the expression of HSP70 in C33A-E6 cells could significantly reduce the secretion of IL-6 and IL-10 by exosome-treated THP-1 cells, and inhibit the THP-1 cell-induced proliferation, migration and invasion of C33A cells ( P<0.001). Conclusions:High-risk HPV 16 E6 protein upregulated the level of HSP70 in the exosomes secreted by cervical cancer cells, thereby promoting the secretion of IL-6 and IL-10 by macrophages and enhancing the macrophage-induced proliferation, migration and invasion of cervical cancer cells.

Brain-computer Interface and Comprehensive Training for Stroke: A Resting State Functional Magnetic Resonance Imaging Study / 中国康复理论与实践

Objective:To investigate the topological alterations in brain functional networks following comprehensive treatment including brain-computer interface (BCI) training in subacute stroke subjects. Methods:From January, 2018 to June, 2019, 14 subacute stroke patients with moderate to severe upper limbs paralysis accepted routine physical therapy, occupational therapy and BCI training based on motor imagery, for four weeks. They were assessed with Fugl-Meyer Assessment-Upper Extremities (FMA-UE), Action Research Arm Test (ARAT) and Wolf Motor Function Test (WMFT) before and after treatment, while the functional connectivity (FC) was investigated with resting state functional magnetic resonance imaging. Results:The scores of FMA-UE, ARAT and WMFT increased after treatment (|t| > 5.298, Z = -3.297, P < 0.01). The FC also increased across the whole brain, including temporal, parietal, occipital lobes and subcortical regions. The FC between left piriform cortex of parietal lobule (BA5L) and right medial surface of temporal lobe (BA48R), as well as those between left precentral gyrus (BA4L) and right anterior transverse temporal gyrus (BA41R) (r > 0.416, P < 0.05). Conclusion:Comprehensive rehabilitation including BCI training may promote recovery of motor function and activities of FC in brain in subacute stroke patients.

Anti-cerebral ischemia mechanisms of baicalein and genipin based on network pharmacology / 中草药

Objective: Using network pharmacology and molecular docking technology along with scutellarein (SE) as a reference, this study predicted the anti-cerebral ischemia targets of both baicalein (BE) and genipin (GE). It is hoped that these will provide a reference for clinical prevention and development of ischemic diseases. Methods: SE, BE and GE targets were predicted using TCMSP, Swiss Target Prediction, Stitch database search and literature mining methods. Targets related to cerebral ischemia diseases could be predicted by DisGeNET, CTD, NCBI Gene, OMIM, DrugBank and PharmGkb databases. Cytoscape 3.3.0 was used to construct the small molecule-target network. GO function enrichment and KEGG pathway analysis of SE, BE and GE specific anti-cerebral ischemia targets were analyzed with the DAVID database. Autodock Vina software was used for molecular docking, testing the binding energy of BE, GE and SE to targets of cerebral ischemia. The optimal target protein was selected according to the binding energy and inhibition concentration of receptor and ligand. Results: A total of 30 potential targets of SE, 59 potential targets of BE and 35 potential targets of GE were found. Common anti-cerebral ischemia targets of SE and BE were PIK3CG, CYP1A2, VEGFA, ALOX5 and PTGS2, while common anti-cerebral ischemia targets of SE and GE were PTGS2. Molecular docking results demonstrated that the binding energy and inhibitory concentration of receptor PTGS2 to the three drugs were relatively low. Enrichment of GO function showed that common targets of BE-SE were mainly distributed in cytoplasm, organelle membrane, endoplasmic reticulum and other elements. These elements had binding functions with metal ions and cations, catalytic and oxidoreductase activities, and they participated in cell lipid, carboxylic acid, oxygenic acid, organic acid metabolism and fatty acid synthesis. Results: of the KEGG analysis demonstrated that receptor PTGS2 mainly acted on the arachidonic acid metabolism pathway and the vascular endothelial growth factor signaling pathway. A combination of BE and GE functioned in the treatment of cerebral ischemia disease by inhibiting expression of PTGS2 (COX-2) and vascular endothelial growth factor protein, thus reducing brain injury caused by inflammatory factors and improving the permeability of the blood brain barrier (BBB). Conclusion: This study predicted potential targets of BE and GE compatibility in the treatment of cerebral ischemia diseases and preliminarily verified mechanisms of action in the treatment of cerebral ischemia diseases. It provides valuable data for further study into the mechanisms of BE and GE compatibility for the treatment of cerebral ischemia as well as developing a basis for the next synthesis of new derivatives.

Research Progress of Traditional Chinese Medicine in Treating Breast Cancer / 中国实验方剂学杂志

Breast cancer is one of the common diseases among women. It is a malignant tumor with a variety of complex mechanisms. Its pathogenesis has not been clearly studied. Physical, chemical and surgical treatments often cause vomiting, nausea, dizziness and headache for women. As compared with traditional treatment, Chinese medicine is characterized by multiple targets, small side effect and good effect in treating breast cancer. In this paper, 85 kinds of Chinese herbal medicines that can treat breast cancer were included. Among them, 69 kinds of Chinese herbal medicines have been included in the 2015 edition of the Chinese Pharmacopoeia, and 16 kinds of Chinese herbal medicines have not been included. The main medicinal ingredients in these Chinese herbal medicines for treatment of breast cancer were alkaloids, glycosides, phenols, terpenes, carbohydrates, volatile oils, coumarins and so on. In addition, these herbal medicines were classified according to their effects in breast cancer. Then, combined with the recent studies at home and abroad, this paper summarized the effect of traditional Chinese medicine(TCM) on breast cancer, including the reversal of multi-drug resistance, the inhibition of metastasis and proliferation, the induction of tumor cell apoptosis, and the arrest of the cell cycle for breast cancer. This paper also explained three pathways for treating breast cancer by TCM, including intervening the tumor cell related apoptosis gene to inhibit breast cancer, inhibiting the expression of P-glycoprotein in the cell membrane to reverse the multi-drug resistance of breast cancer cells, and regulating the related epithelial mesenchymal transition signal pathway to prevent breast cancer cells metastasis and proliferation. In the end, it was concluded that Chinese medicine can reduce the drug resistance and metastasis of breast cancer cells, block the cell cycle of breast cancer cells, and also intervene the expression of apoptotic factors to promote the death of breast cancer cells. The inhibition of breast cancer by Chinese medicine was the result of the common effect of various ingredients. Therefore, Chinese medicine treatment for breast cancer has the unparalleled advantages as compared with chemical and surgical treatment. Chinese medicine is one of our important means to overcome breast cancer.

The key issues and corresponding strategy of acupoint medicated catgut embedding therapy / 中国针灸

Acupoint medicated catgut embedding therapy is a new type of acupuncture combined with medicine，which has great potential in the prevention and treatment of chronic diseases. This paper describes the clinical application of acupoint medicated catgut embedding in the treatment of common disease, medicated thread productionmethods and acupoints selection.To analyze the problems of the unclear mechanism, the lack of standardization of medicated thread production, the lag of the embedding tool, and the unclear adverse reaction. It is expected to promote the further development of acupoint medicated catgut embedding therapy by strengthening the mechanism of action, optimizing medicated catgut thread, rational design experiments, standardizing medicated thread production, innovating embedding tools, rational formulation, and optimizing acupoint selection.