Abstract: Objective　To investigate the relationship between the body mass index (BMI) levels and the negative conversion time of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleic acid in adult coronavirus disease 2019 (COVID-19) patients and the asymptomatic persons. Methods　Asymptomatic infected patients and confirmed COVID-19 patients admitted to Chengdu Public Health Clinic Center from February 2021 to November 2021 were dynamically included. The epidemiological and clinical characteristics of the objects were collected, and the SARS-CoV-2 nucleic acid testing of the objects during their hospitalization was continuously monitored, and the negative nucleic acid conversion time was recorded. The t test or Wilcoxon rank sum test, χ2 test or Fisher's exact probability method examine were used to distribute characteristics of each group of variables and the connection between different variables, respectively. Then the variables showed differences in distribution (P<0.05) between different BMI groups were included in the multivariate Cox proportional risk regression model. Results　A total of 253 subjects ranged from 18 to 63 years old, with M(P25, P75) age of 37.0 (30.0, 47.0) years old, were included in this study. The male to female ratio was 4.16 to 1. The BMI was (23.97±3.33) kg/m2. 50.59% (128/253) of the objects were overweight or obese, and 78.13% (100/128) were overweight. The negative time of SARS-CoV-2 nucleic acid conversion of all subjects ranged from 1 to 71 days, with M(P25, P75) of 7.0 (2.0, 18.0) days (P<0.001). The negative time of SARS-CoV-2 nucleic acid conversion of the normal weight or the thin, and the overweight or obese were 5.00 (2.00, 19.00) and 8.00 (2.00, 17.75) days respectively. The results of multivariate Cox's proportional hazards regression model showed that the BMI levels may not be associated with the negative conversion time of SARS-CoV-2 nucleic acid (HR=1.090, 95%CI: 0.843-1.410, P=0.510). Conclusions　Adult asymptomatic persons and confirmed COVID-19 patients are mainly middle-aged and young males, and overweight or obesity is relatively common. Overweight or obesity cannot be considered as an independent factor influencing the negative conversion time of SARS-CoV-2 nucleic acid.

This study is to evaluate the therapeutic effect of fibroblast growth factor 21 (FGF21) on type 2 diabetic mice model and to provide mechanistic insights into its therapeutic effect. Type 2 diabetic animal model was established with high calorie fat diet and low dose streptozotocin (STZ) injection. Mice were then randomized into 5 groups: model control, FGF21 0.25 and 0.05 μmol x kg(-1) x d(-1) groups, insulin treatment group. Ten age-matched normal KM mouse administered with saline were used as normal controls. Serum glucose, insulin, lipid products and the change of serum and liver tissue inflammation factor levels between five groups of mouse were determined. The results showed that blood glucose, insulin, free fatty acids (FFAs), triglycerides, and inflammatory factor average FGF-21 of type 2 diabetes model group and normal control group were significantly higher (P < 0.01), while compared with insulin group, no difference was significant. Average blood glucose, insulin, blood lipid and inflammatory factor of FGF-21 treatment group compared with type 2 diabetes group was significantly lower (P < 0.01) and insulin group has no difference with the model control group. The results of OGTT and HOMA-IR showed that insulin resistance state was significantly relieved in a dose-dependent manner. Thus, this study demonstrates that FGF-21 significantly remits type 2 diabetic mice model's insulin resistance state and participates in the regulation of inflammatory factor levels and type 2 diabetes metabolic disorders.
Animals ; Blood Glucose ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetes Mellitus, Type 2 ; drug therapy ; Diet, High-Fat ; Fatty Acids, Nonesterified ; blood ; Fibroblast Growth Factors ; pharmacology ; Insulin ; blood ; Insulin Resistance ; Mice ; Streptozocin ; Triglycerides ; blood

The ZM-1 tissue microarrayer designed by our groups is manufactured in stainless steel and brass and contains many features that make TMA (tissue microarray) paraffin blocks construction faster and more convenient. By means of ZM-1 tissue microarrayer, biopsy needles are used to punch the donor tissue specimens respectively. All the needles with the punched specimen cylinders are arrayed into the array-board, with an array of small holes dug to fit the needles. All the specimen cylinders arraying and the TMA paraffin block shaping are finished in only one step so that the specimen cylinders and the paraffin of the TMA block can very easily be incorporated and the recipient paraffin blocks need not be made in advance, and the paraffin used is the same as that for conventional pathology purpose. ZM-1 tissue microarrayer is easy to be manufactured, does not need any precision location system, and so is much cheaper than the currently used instrument. Our method's relatively cheap and simple ZM-1 tissue microarrayer technique of constructing TMA paraffin block may facilitate popularization of the TMA technology.
Biopsy, Needle ; instrumentation ; Equipment Design ; Female ; Humans ; Immunohistochemistry ; Male ; Neoplasms ; enzymology ; Paraffin ; Tissue Array Analysis ; instrumentation

OBJECTIVETo investigate the effect of rat mesenchymal stem cells (MSCs) transfected with different oncogenes differentiate into hepatocellular carcinoma (HCC) in vitro.

METHODSMSCs, transfected with different oncogenes c-myc, K-ras, c-myc and K-ras and amplified in vitro, were infused into rats via vena portae. The recipient rats were divided into the hepatic impairment group, which were fed with tetrachloromethane and the healthy control group. At day 7, 14, 21, 28 and 42 following grafting, the complete livers were obtained and examined using fluorescence detection, conventional pathology and immunohistochemistry detection of GFP, c-kit and AFP to study the colonization and distribution of stem cells in rat liver.

RESULTSNo immunological rejection occurred after grafting of allogenic MSCs. The infused MSCs colonized in the recipient rat liver. Liver tumors were present in 6 rats grafted with MSCs that were transfected with K-ras, K-ras and c-myc, and the protein expression of AFP was detected using immunocytochemistry at day 7. Rats grafted with MSCs that were transfected with c-myc gene had no obvious tuberosity or tumor. Small oval cells were found microscopically in the periphery of vena portae, and immunohistochemistry staining of AFP was negative. Immunohistochemical staining of c-kit was positive in all livers of rats that were transfected with MSCs.

CONCLUSIONSHepatocellular carcinoma may derive from genetically mutated MSCs.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Differentiation ; Cells, Cultured ; Female ; Genes, myc ; genetics ; Genes, ras ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Liver ; metabolism ; pathology ; Liver Neoplasms, Experimental ; metabolism ; pathology ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Microscopy, Fluorescence ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo study the relationship between extrahepatic metastasis of primary hepatocellular carcinoma and circulative tumor cells in the blood of hepatoma patients.

METHODSThe immunomagnetic bead technique was employed to enrich and separate the hepatoma cells in the peripheral blood of preoperative and postoperative hepatoma patients. The relationship between postoperative extrahepatic metastasis and hepatoma cells in peripheral blood cancer cells were analyzed. The circulative tumor cells in the peripheral blood of hepatoma patients were enriched and separated by immunomagnetic bead technique. They were identified as hepatoma cells by AFP immunohistochemistry. Among 30 cases of hepatoma patients, the positive rate of hepatoma cells in the peripheral blood of preoperation and postoperation were 53.3% and 83.3% respectively. There was difference significantly in positive cases before operation and after operation (P<0.05).

CONCLUSIONSExtrahepatic metastasis of primary hepatocellular carcinoma is obviously correlated to the positive tumor cells and the concentration in the peripheral blood of preoperative patients.

Adult ; Carcinoma, Hepatocellular ; blood ; pathology ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms ; blood ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging

ZM-1 tissue microarrayer designed by our group is manufactured in stainless steel and brass. It features an easier and faster preparation for tissue microarrays. By means of it, a group of biopsy needles are used to punch the donor tissue specimens respectively, and all the needles with the punched specimen cylinders are arranged into the array-board, where small holes have been digged to fit the needles. All the specimen cylinders arraying and the tissue microarray block's shaping are finished simultaneously. ZM-1 tissue microarrayer with a lower cost of manufacture, is capable of preparing the tissue microarrays conveniently, efficiently and quality-controllably.
Equipment Design ; Tissue Array Analysis ; instrumentation

Objective To etiologically diagnose and analyze a patient with suspected cases of brucellosis,and to provide a experimental basis for the confirmation of the first case of human brucellosis in Guizhou province.Methods Conventional and molecular techniques [genus specific Brucella surface protein 31 PCR (BCSP31-PCR)and Brucella suis species-specific PCR (AMOS-PCR)] were used to identify suspicious bacteria strains isolated from the suspected patient of brucellosis.Results The results showed that the Brucella suspicious colonies were identified as Brucella melitensis biotype 3 using conventional tests and were further identified as Brucella spp.by genus specific Brucella surface protein 31 PCR (BCSP31-PCR) and classified as Brucella melitensis with Brucella abortus,Brucella melitensis,Brucella ovis,Brucella suis species-specific PCR(AMOS-PCR).Conclusions Laboratory diagnostic results show that the bacteria strain isolated from the suspected patient of brucellosis is Brucella melitensis biotype 3.It is the first case of human brucellosis in Guizhou province.

Objective To evaluate the value of positron emission tomography(PET)-CT imaging combined with continual detection of CA_(125)in serum for diagnosis of early recurrent ovarian epithelial carcinoma.Methods Twenty six patients received PET-CT imaging,who were all diagnosed as primary epithelial ovarian cancer of stage Ⅱ-Ⅳ and had complete remission after cytoreductive surgery and multiple courses of chemotherapy in Shandong Provincial Cancer Hospital.After a steady period,all patients experienced progressive rising of CA_(125)values 3 times in 2 months.But no positive lesion was found by CT, or although suspicious positive focus was found,the recurrent and(or)metastatic extent was not definite. Out of 26 patients,16 were delivered rechemotherapy and(or)radiotherapy,and 10 received re- cytoreductive surgery.Results(1)Of 26 patients,the value of CA_(125)was more than 35 kU/L in 17,and in 14 of 17,pelvic or abdominal cavity recurrence was diagnosed by CT and PET-CT,and 4 showed simuhaneously distant metastasis on PET-CT.In the remaining 3 patients of which CT findings were negative,2 had pelvic and abdominal cavity recurrence,and one had bone metastasis on PET-CT.Of 9 patients with progressive rising CA_(125)levels but the value was less than cut-off(

OBJECTIVETo assay the expression of KiSS-1 and GnRH in the male rat hypothalamus at different developmental stages, and to explore the significance of KiSS-1 in sex development onset and normal reproduction regulation.

METHODSExpression analyses of KiSS-1 and GnRH genes were conducted in the rat hypothalamus at different developmental stages with RT-PCR and real time-PCR. The testosterone level was assayed by chemoluminescence technique.

RESULTSKiSS-1 mRNA rose gradually during sex development in the rat hypothalamus, highest at puberty and lowered a little at adulthood. KiSS-1 mRNA of the prepubertal, early pubertal, pubertal and adult rats was 1.7, 2.1, 3.5 and 2.0 times higher than that of the infantile rats respectively. The expression of GnRH and KiSS-1 correlated positively (r = 0.905, P < 0.05). But the activation of GnRH neuron was later than KiSS-1. The expression of GnRH was the highest in the puberty rats. GnRH mRNA of the prepubertal, early pubertal, pubertal and adult rats was 1.1, 1.94, 2.42 and 1.92 times higher than that of the infantile rats respectively. The level of testosterone in the adult rats was significantly higher than that at the earlier stage and was the highest at the adult stage.

CONCLUSIONThe expression of KiSS-1 correlates positively with that of GnRH. KiSS-1 may participate in the regulation of GnRH and is relevant to puberty onset and the regulation of reproduction function.

Animals ; Gonadotropin-Releasing Hormone ; biosynthesis ; genetics ; Hypothalamus ; metabolism ; Kisspeptins ; Male ; Proteins ; metabolism ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

The full length of chicken interferon alpha (ChIFN-alpha) gene was amplified by the polymerase chain reaction (PCR) from total liver genome of Sanhuang meat-chicken and sequenced. The amplified gene was about 582bp. The coding region for mature protein (489bp) was subcloned into pET-28a(+). The recombinant plasmid pET-28a(+)-IFNalpha was identified by enzyme digestion and DNA sequencing. Data of SDS-PAGE and Western-blot indicated that a 22kD fusion protein was expressed in the form of inclusion bodies with good immunity. The purity of inclusion bodies was above 70% and that of protein purified by nickel affinity chromatography was 95%. The recombinant protein could inhibit H9N2 avian influenza virus (H9N2 AIV) replication on chick embryo fibroblast. 2 microg of recombinant IFN-alpha could completely protect Chick embryo from H9N2 AIV infection. The recombinant IFN-alpha can also delay Newcastle disease virus (NDV) replication on chick embryo for 12 approximately 48h. Chicken administered recombinant IFN-alpha can resist the H9N2 AIV infection. The bioactivities of recombinant IFN-alpha purified by affinity chromatograph were 20 times higher than that of inclusion bodies.
Animals ; Antiviral Agents ; pharmacology ; Blotting, Western ; Chick Embryo ; Cloning, Molecular ; Influenza A Virus, H9N2 Subtype ; drug effects ; Interferon Type I ; biosynthesis ; isolation & purification ; pharmacology ; Interferon-alpha ; genetics ; Newcastle disease virus ; drug effects ; Plasmids ; Recombinant Proteins