Radix Scrophulariae Extracts Exert Effect on Hyperthyroidism via MST1/Hippo Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To explore the mechanism of Radix Scrophulariae (RS) extracts in the treatment of hyperthyroidism rats by regulating proliferation, apoptosis, and autophagy of thyroid cell through the mammalian sterile 20-like kinase 1 (MST1)/Hippo pathway.@*METHODS@#Twenty-four rats were randomly divided into 4 groups according to a random number table: control, model group, RS, and RS+Hippo inhibitor (XMU-MP-1) groups (n=6 per group). Rats were gavaged with levothyroxine sodium tablet suspension (LST, 8 μ g/kg) for 21 days except for the control group. Afterwards, rats in the RS group were gavaged with RS extracts at the dose of 1,350 mg/kg, and rats in the RS+XMU-MP-1 group were gavaged with 1,350 mg/kg RS extracts and 1 mg/kg XMU-MP-1. After 15 days of administration, thyroid gland was taken for gross observation, and histopathological changes were observed by hematoxylin-eosin staining. The structure of Golgi secretory vesicles in thyroid tissues was observed by transmission electron microscopy. The expression of thyrotropin receptor (TSH-R) was observed by immunohistochemistry. Terminal-deoxynucleoitidyl transferase mediated nick end labeling assay was used to detect cell apoptosis in thyroid tissues. Real-time quantity primer chain reaction and Western blot were used to detect the expressions of MST1, p-large tumor suppressor gene 1 (LATS1), p-Yes1 associated transcriptional regulator (YAP), proliferating cell nuclear antigen (PCNA), G1/S-specific cyclin-D1 (Cyclin D1), B-cell lymphoma-2 (Bcl-2), Caspase-3, microtubule-associated proeins light chain 3 II/I (LC3-II/I), and recombinant human autophagy related 5 (ATG5). Thyroxine (T4) level was detected by enzyme-linked immunosorbent assay.@*RESULTS@#The thyroid volume of rats in the model group was significantly increased compared to the normal control group (P<0.01), and pathological changes such as uneven size of follicular epithelial cells, disorderly arrangement, and irregular morphology occurred. The secretion of small vesicles by Golgi apparatus was reduced, and the expressions of receptor protein TSH-R and T4 were significantly increased (P<0.01), while the expressions of MST1, p-LATS1, p-YAP, Caspase-3, LC3-II/I, and ATG5 were significantly decreased (P<0.01). The expressions of Bcl-2, PCNA, and cyclin D1 were significantly increased (P<0.01). Compared with the model group, RS extracts reduced the volume of thyroid gland, improved pathological condition of the thyroid gland, promoted secretion of the secretory vesicles with double-layer membrane structure in thyroid Golgi, significantly inhibited the expression of TSH-R and T4 levels (P<0.01), upregulated MST1, p-LATS1, p-YAP, Caspase-3, LC3-II/I, and ATG5 expressions (P<0.01), and downregulated Bcl-2, PCNA, and Cyclin D1 expressions (P<0.01). XMU-MP-1 inhibited the intervention effects of RS extracts (P<0.01).@*CONCLUSION@#RS extracts could inhibit proliferation and promote apoptosis and autophagy in thyroid tissues through MST1/Hippo pathway for treating hyperthyroidism.

Rates and influencing factors of hospitalization after diagnosis among HIV infection cases in Yinzhou district of Ningbo, 2012-2020 / 中华流行病学杂志

Objective: To understand the hospitalization rates and influencing factors after diagnosis among HIV infection cases, based on real-world data in Yinzhou district of Ningbo. Methods: A retrospective cohort study was conducted based on the databases of National AIDS Comprehensive Response Information Management System and Yinzhou Health Information Platform. The information about the following-up results, antiviral treatment data, electronic records of inpatient of the HIV cases reported during 2012-2020 were collected to analyze the rates, causes and influencing factors of hospitalization. Results: Among the 763 HIV infection cases reported in Yinzhou from 2012 to 2020, the hospitalization rate was 6.95% (53/763), and the number of inpatient was 2.59 per 100 person years. The hospitalization rate and the number of hospitalization per 100 person years in HIV infection cases were 3.16% (10/316) and 0.81 in those aged <30 years, 6.07% (15/247) and 1.59 in those aged >30 years, 7.86% (11/140) and 4.05 in those aged >45 years and 28.33% (17/60) and 17.40 in those aged ≥60 years respectively. Logistic multivariate regression analysis indicated that being aged ≥60 years was the influencing factor for hospitalizations in HIV infection cases (аOR=14.44, 95%CI:3.57-58.46). The hospitalization rates due to AIDS related diseases, cardiovascular diseases and metabolic diseases, and other diseases were 1.83% (14/763), 1.05% (8/763), and 3.93% (30/763), respectively. Conclusions: The hospitalization burden due to HIV infection was still mainly caused by those aged ≥60 years in Yinzhou, similar to that in general population and less proportion of hospitalizations were due to AIDS related diseases. The overall increase of hospitalizations due to AIDS was not obvious in Yinzhou.

Development of an early warning index system for emerging respiratory infectious diseases in Shanghai / 上海预防医学

Objective:To develop an early warning index system to identify the outbreak of respiratory infectious diseases and to assess the epidemic risk of these diseases in Shanghai. Methods:A two-round Delphi survey with a panel of 31 experts was used to select the modifying indicators. All indicators were evaluated for necessity，feasibility，stability， and sensitivity through online questionnaires. Results:The consultation recovery rates of both rounds were 100%. The authority coefficient was 0.88， and the Kendall's W of the second-round consultation was 0.138 （P<0.001）， suggesting a good coordination among experts’ opinion. Our early warning index system was developed after the two-round Delphi survey and included two sets of indicators， one for rapid risk assessment and another for monitoring and tracing risk. Both sets of the indicators involved three aspects： transmissibility， clinical severity and potential influence of emerging diseases. The rapid risk assessment system part covered 10 indicators while the risk monitor system part covered 23 indicators. In the rapid risk assessment system part， the weight value of the top three indicators were greater than 0.10， with the highest weight value of 0.171 for titled visiting rate of influenza-like illness patients in the Emergency Department or other outpatient visits. In the risk monitor system part， the weight value of the top eight indicators were greater than 0.05， and the greatest was 0.087 for R0 （basic reproductive number）. Conclusion:The developed scientific and reliable evaluation indicator system can be used to forecast the outbreak and epidemic risk of respiratory infectious diseases. However， sensitivity and stability of the indicators need further validation and evaluation.

Effect of Shenling Baizhusan on Protein and mRNA Expression of NF-κB p65，IκBα，IκKβ in Ulcerative Colitis Rats with Syndrome of Dampness Stagnancy Due to Spleen Deficiency / 中国实验方剂学杂志

Objective:To observe the effect of Shenling Baizhusan on the protein and mRNA expression of inhibitor of nuclear factor kappa B kinase (IκK)/inhibitor of nuclear factor kappa B(IκB)/nuclear factor kappa B(NF-κB) signaling pathway in the colon of rats with ulcerative colitis (UC) of spleen deficiency and dampness stagnation type, and to explore the mechanism of Shenling Baizhusan in the treatment of UC. Method:The 48 Wistar rats were randomly divided into normal group, model group, Shenling Baizhusan group (15.6 g·kg-1) and osalazine sodium group (0.68 g·kg-1), 12 rats in each group. The model of UC with spleen deficiency and dampness stagnation was reproduced by trinitrobenzene sulfonic acid (TNBS)/ethanol enema combined with environment and diet intervention.Serum levels of tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β(IL-1β) were determined by enzyme-linked immunosorbent assay（ELISA）. The expression of NF-κB p65, IκBα, IκKβ protein in colon tissue was measured by Western blot and immunohistochemical method, and the mRNA expression of NF-κB p65, IκBα and IκKβ in colon tissue of rats in each group was detected and compared by real time polymerase chain reaction (Real-time PCR). Result:Compared with normal group,the levels of TNF-α,IL-6 and IL-1β in the serum, the protein and mRNA expression of NF-κB p65, IκKβ in colon tissue of the model group was significantly higher than that of normal group (P<0.01), and the protein and mRNA expression of IκBα was significantly lower than that of normal group (P<0.01). Compared with model group,the protein and mRNA expression of NF-κB p65, IκKβ in colon tissue of the Shenling Baizhusan group and osalazine sodium group were significantly decreased (P<0.01), and the protein and mRNA expression of IκBα was significantly increased (P<0.01). Conclusion:Shenling Baizhusan can obviously down regulate the protein and mRNA expression of NF-κB p65, IκKβ,up regulate the expression of IκBα in colon tissue of UC rats with spleen deficiency and dampness stagnation. The inhibition of IκK/IκB/NF-κB signal pathway activation by Shenling Baizhusan is an important mechanism of its role in protecting intestinal mucosa.

Analysis of Effect of Chaihu Longgu Mulitang on Intestinal Microflora Diversity of Schizophrenic Rats Based on 16S rRNA Technique / 中国实验方剂学杂志

Objective:To explore the effect of Chaihu Longgu Mulitang on intestinal microflora diversity of schizophrenic model rats, and further reveal its therapeutic characteristics and mechanisms based on the 16S rRNA technique. Method:Except the normal group, male SD rats were intraperitoneally injected dizocilpine maleate with daily dose of 0.1 mg·kg-1.After the success of the model, Chaihu Longgu Mulitang high, middle and low dose groups were converted into the human clinical upper limit daily, and the experimental rats were given Chaihu Longgu Mulitang with doses of 11.2, 5.6, 2.8 g·kg-1, respectively. And the positive drug group was treated with 0.4 mg·kg-1 of risperidone tablets.The normal group and model group was treated with water.The rats were continuous administrated 14 days with dosing volume of 10 mL·kg-1, the contents in caecum of rats were taken after anesthesia.Illumina MiSeq was used as the sequencing platform, the number of operational taxonomic units(OTUs), richness and diversity indexes, diversity of alpha and beta, differential phylum and genus of intestinal flora in V4 zone of 16S rRNA were comprehensively analyzed and evaluated. Result:Chaihu Longgu Mulitang could improve the number of OTUs, richness and diversity indexes of intestinal flora, imbalance of alpha and beta diversity of schizophrenic model rats.And this formula had a callback effect on 5 differential phyla of bacteria(Bacteroidetes, Acidobacteria, Proteobacteria, Firmicutes and TM7) and 20 genera of bacteria in schizophrenic model rats. Conclusion:Chaihu Longgu Mulitang plays an therapeutic effect on diversity of abnormal microflora in schizophrenic model rats, and this paper reveals the pathological mechanism of intestinal microflora in the state of schizophrenia by 16S rRNA technique.

Intestinal Microflora Diversity of Rats Treated with Scutellariae Radix Based on Theory of “Insight into Abdomen” / 中国实验方剂学杂志

Objective: To explore the effect of Scutellariae Radix on the diversity of intestinal flora in rats under physiological conditions, in order to determine the property of Scutellariae Radix property. Method: 16S rRNA high-throughput gene sequencing technique was used to detect the cecum solutes of rats treated with Scutellariae Radix (10 g·kg-1). The number, richness and diversity index of the intestinal flora taxon (OTUs) and the differential phylum and genus were comprehensively analyzed. The network visualization was used to find the correlation between differential phylum and genus. Result: Based on the Illumina Miseq platform, compared with the blank group, the number of OTUs and the index of richness and diversity of the intestinal flora of the rats treated with Scutellariae Radix decreased. Bacteroidetes and Spirochaetes were significantly up-regulated(PPPPPRuminococcus, Paraprevotella, Prevotella and Oscillospira. Conclusion: Scutellariae Radix can reduce the diversity of intestinal flora and inhibit the metabolism of the body, so its property is cold.

Comparison of sagittal plane morphology of spine and pelvis in adolescents with L₅S₁ developmental spondylolisthesis and isthmic spondylolisthesis / 中国骨伤

OBJECTIVE@#To compare the sagittal morphological features of the spine and pelvis between L₅S₁ dysplastic spondylolisthesis and isthmus spondylolisthesis in adolescent.@*METHODS@#Retrospective analysis of 24 cases of adolescent L₅S₁ spondylolisthesis with complete imaging data from May 2002 to December 2016. Those included 8 males and 16 females, aged from 10 to 18 years old with an average of (13.4±2.0) years. Among them, 9 cases were diagnosed as dysplastic spondylolisthesis (dysplasia group) and 15 cases isthmic spondylolisthesis (ischemic group). Radiographic parameters including slippage distance, slippage degree, slippage angle, sagittal vertical axis(SVA), thoracic kyphosis(TK), lumbar lordosis(LL), L₅ incidence(L₅I), pelvic incidence(PI), pelvic tilt(PT), sacral slope(SS), sagittal pelvic thickness(SPT), lumbosacral angle (LSA), sacral table angle (STA) were measured on the spinal lateral X-ray of the standing position. Independent-samples t-test was used in the comparison of each variable between two groups. 0.05).@*CONCLUSIONS@#Significant different from isthmic spondylolisthesis, adolescents with dysplastic spondylolisthesis present a different spino-pelvic sagittal alignment, characterized with trunk forward leaning and pelvic retroversion. In case of sagittal imbalance, early surgical intervention is required to restore a balanced spino-pelvic alignment.

Study on Radix Scrophulariae for the treatment of hyperthyroidism rat model of yin deficiency based on urine metabonomics / 药学学报

The Chinese herbal Radix Scrophulariae is the main medicine for nourishing yin and reducing fire. It can be used to treat hyperthyroidism due to yin deficiency and fire hyperactivity, but its mechanism is not clear. The present study was aimed to explore the mechanism of Radix Scrophulariae treatment of hyperthyroidism due to yin deficiency and fire hyperactivity. The urine metabolomic approach was conducted using the method of UPLC-TOF-MS. The results showed that Radix Scrophulariae has good therapeutic effects on hyperthyroidism rat model of yin deficiency. After treatment with Radix Scrophulariae, through metabolic profiling and protocol analysis, 6 potential metabolic markers may be closely related with the treatment mechanism of Radix Scrophulariae on this disease, including proline betaine, estrone, thymidine, 5-hydroxyindoleacetic acid, cyclic AMP and L-dopa. The strongest metabolic pathways were associated with tryptophan metabolism, pyrimidine metabolism, tyrosine metabolism, purine metabolism and steroid hormone biosynthesis. The urine metabolomic approach can be applied to clarify the therapeutic mechanism of Radix Scrophulariae on hyperthyroidism rat of yin deficiency, and provide the theoretical basis for the clinical practice of Radix Scrophulariae on nourishing yin to reduce pathogenic fire.

Construction of pEGFP-C1-CXCL1 plasmid and detection of its expression in eukaryotic cells / 军事医学

Objective To construct the pEGFP-C1-CXCL1 eukaryotic expression vector and to investigate the effect of CXCL1 on the proliferation of HepG2 cells under endoplasmic reticulum stress ( ERS).Methods Fragments of CXCL1 were obtained from the cDNA library of HepG2 cells before CXCL1 was cloned into a pEGFP-C1 vector for a recombinant plasmid pEGFP-C1-CXCL1 which was screened and identified by PCR and sequence alignment .Then,the recombinant plas-mid of pEGFP-C1-CXCL1 was transfected into human 293 T cell line and the expression of CXCL 1 was detected by fluores-cence microscopy and Western blotting.pEGFP-C1-CXCL1was furhter transfected into HepG2 cells, and CCK8 was used to detect the inhibitory effect of CXCL1 on tumor proliferation induced by TM in hepatocellular carcinoma .Results pEGFP-C1-CXCL1 was vertified by sequencing analysis .Fluorescence microscopy showed that pEGFP-C1-CXCL1 was transfected into 293T.CXCL1 expression was detected by Western blotting .CCK8 showed that TM inhibited tumor proliferation , while overexpression of CXCL1 decreased the inhabitory rate on cell proliferation of HepG 2 cells under ER stress compared to pEGFP-C1 group and the control group .Conclusion A recombinant pEGFP-C1-CXCL1 plasmid is successfully constructed that can be expressed stably in human 293T cells.Overexpression of CXCL1 can effectively reduce the inhabitory rate of HCC cells induced by the ER stress.

Prokaryotic expression, purification, and immunogenicity analysis of Mycobacterium tuberculosis specific excretive proteins / 中国医学科学院学报

<p><b>OBJECTIVE</b>To obtain the recombinant rv1837c and rv3803c of Mycobacterium tuberculosis using gene engineering technology and explore their prokaryotic expression, purification, and immunogenicity.</p><p><b>METHODS</b>The Mycobacterium tuberculosis rv1837c and rv3803c genes were amplified by polymerase chain reaction, and then cloned into the vector pTA2, followed by the subclone into the expression vector pET30a (+). The resulting plasmids, named pET30a (+): rv1837c and pET30a (+): rv3803c, encode recombinant protein containing a hexa-histidine tag on its N-terminus. pET30a (+): rv1837c and pET30a (+): rv3803c were introduced into E. coli BL21 (DE3) by transformation respectively, and the recombinant gene was induced with 0.4 mmol/L isopropyl-D-thiogalactopyranoside. The expressed products were identified by Western blot with hexa-histidine tag antibody and serum from tuberculotic patients. The histidine tagged protein was purified by nickel nitrilotriacetic acid His-Bind resin. Rabbits were immunized with purified recombinant Rv1837c and Rv3803c proteins. Then the purified recombinant Rv1837c and Rv3803c proteins were used to detect antibody in rabbit serum, which had been immunized by Western blot.</p><p><b>RESULTS</b>After transformation of the E. coli and induction with 0.4 mmol/L of isopropyl-D-thiogalactopyranoside, recombinant target proteins Rv1837c (relative molecular mass: 92000) and Rv3803c (relative molecular mass: 38 000) were expressed in pET30a (+): rv1837c and pET30a (+): rv3803c system. The expressed protein existed in cytoplasm in an unsoluble form and amounted to 30% and 50% of the total proteins of E. coli. The purity of the purified protein reached 90%. The immunogenicity of the recombinant proteins Rv1837c and Rv3803c was strong, as identified by Western blot.</p><p><b>CONCLUSION</b>The prokaryotic expression recombinant plasmids pET30a (+): rv1837c and pET30a (+): rv3803c was successfully constructed and the recombinant proteins Rv1837c and Rv3803c were obtained, which laid a basis for the optimized diagnosis of active tuberculosis.</p>