OBJECTIVE:To evaluate the efficacy and safety of salmon calcitonin plus Xianling gubao for osteoporosis and ostealgia in postmenopausal women. METHODS:160 women with postmenopausal osteoporosis and ostealgia were randomized to 3 groups (treatment group and 2 control groups):the treatment group received(500 IU q.d) salmon calcitonin nasal spray plus Xianling gubao(1.5 g b.i.d orally); the control group Ⅰ received(500 IU q.d) salmon calcitonin nasal spray alone and the control group Ⅱ received Xianling gubao(1.5 g b.i.d orally) alone. All the patients received oral Caltrate D (1 tablet q.d). A course was defined as 30 days. After treatment for a total of 3 courses,the nature and degree of ostealgia in all the patients were assessed,and the outcome indexes were measured and the adverse drug reactions were recorded. RESULTS:There were significant differences between the treatment group and two control groups in total effective rates(92.59% for treatment group vs. 69.81% for control group Ⅰ and 67.92% for control group Ⅱ,P0.05). CONCLUSION:Treatment of osteoporosis and ostealgia in postmenopausal women with salmon calcitonin plus Xianling gubao has been proved to be safe and effective in that the bone density can be increased effectively and the osteoporotic pain of lumbar and back of the patients can be relieved.

Adolescent ; Adult ; Athletic Injuries ; Clinical Clerkship ; Female ; Humans ; Nurses ; Occupational Injuries ; Young Adult

China ; Gastroesophageal Reflux ; therapy ; Humans ; Integrative Medicine ; Medicine, Chinese Traditional ; methods

Objective To explore the possible effect of combined treatment of zoledronic acid and doxorubicin on proliferation,invasion and adhesion of SGC7901 cell line,and potential underlying mechanisms. Methods MTT and Transwell experiments were used respectively to investigate the effect of different concentrations of zoledronic acid and doxorubicin on the proliferation,adhesion and invasion of SGC7901. The mRNA levels of MMP2, TIMP2, MMP9, ICAM1 and CD44 were quantified by real time PCR. Results Gastric cell line SGC-7901 were incubated with zoledronic acid for 48 h in different concentration of 10-6,10-5,10-4,10-3 mol/L,their inhibition rates were 4.98％,12.19％,27.34％ and 73.13％ separately,which were higher than control group (P＜0.01),and enhanced along with the dose increasing and the time extension (P＜0.05).Gastric cell line SGC-7901 was incubated with zoledronic acid 10-4 mol /L and doxorubicin 0.5 mg/L,respectively,or in combination for 2 h,4 h,6 h,and thereafter,the detected adhesion rates were 5.03％,24.38％ and 59.65％ in control group,4.40％,19.77％ and 51.22％ in zoledronic acid group,4.28％, 18.62％ and 51.02％ in doxorubicin group,4.02 ％,15.02％ and 46.95％ in combination group. The adhesion rates were reduced after incubation with different medicament concentration (P＜0.05).In the study of cell invasion,the cell numbers per high power field were:105.11 ± 10.43,98.37 ± 15.75,96.19 ± 9.24,83.02 ± 19.72 after treatment with zoledronic acid,doxorubicin or their combination at different concentrations. Compared with control, signal medicament had weak effectiveness in invasiveness in SGC-7901 (P＞ 0.05),whereas in combination group it was significantly decreased (P＜0.05).Both zoledronic acid and doxorubicin decreased the mRNA expressions of CD44,MMP2,MMP9,ICAM1 while increased TIMP2 mRNA level (P＜0.05or P＜0.01) versus control.The effect was more evident with combination treatment(P＜0.05 or P＜0.01),compared with doxorubicin group. Conclusions Both zoledronic acid and doxorubicin can inhibit the proliferation,adhesion and invasion of SGC7901. Combination treatment has additive effect.It may be related to the down-regulation of mRNA of ICAM1,MMP2,MMP9,CD44 and up-regulation of mRNA of TIMP2.

OBJECTIVETo observe mainfestations of syndrome and biochemical indices of hypertensive model rats with excessive accumulation of phlegm-dampness syndrome (EAPDS), and to explore its possible pathological mechanism.

METHODSEAPDS rat model was prepared in 50 Wistar rats by feeding with high fat forage. Meanwhile, a normal control group consisting of 10 Wistar rats was set up by feeding with normal forage. After 25-week continuous feeding, 22 rats with body weight (BW) and blood pressure (BP) exceeding 25% those of the control group were selected as a model group. BW, BP, blood lipids, and related serological indicators were detected in all rats. Morphological changes of target organs were observed. mRNA expression levels of leptin receptor (LepR), Janus kinase2 (Jak2), signal transducer and activator of transcription 3 (Stat3), suppressor of cytokine signaling-3 (Socs3), angiotensin II receptor type 1 (AT1), angiotensin II receptor type 2 (AT2), phosphatidylinositol 3 kinase (P13K), serine threonine kinase (Akt), nuclear factor of kappa B (NF-κBp65), inhibitor of nuclear factor kappa-B kinase α (IKKα), NF-kappa-B inhibitor β (lKKβ), NF-kappa-B inhibitor α (IKBα), and AMP-activated protein kinase (AMPK) were detected by quantitative real-time PCR (qPCR). Expression levels of AT1 and LepR in aorta were detected by immunohistochemical assay and Western blot respectively.

RESULTSCompared with the control group, BW, BP, and blood lipids increased; serum levels of leptin (Lep) , Ang II, Hcy, ET-1, TNF-α, IL-6, and p2-MG increased, but NO decreased in the model group (P < 0.05, P < 0.01). Aortal endothelial injury and smooth muscle cell proliferation occurred in the model group, accompanied with heart and renal injury. Compared with the control group, mRNA expression levels of LepR, Jak2, Stat3, Socs3, AT1 , PI3K, Akt, NF-κB p65, IKKβ, IKBα, and AMPK in aorta were up-regulated significantly (P < 0.05), while the expression of IKKa decreased (P < 0.05). Immunohistochem- ical staining showed, brownish yellow deposit of AT1 and LepR was obviously increased, with more extensively positive distribution. Western blot results showed, as compared with the control group, protein expression levels of AT1 and LepR obviously increased in the model group (P < 0.05).

CONCLUSIONSModel rats exhibited typical syndromes of EAPDS. They put up weight with fat abdomen, gloomy hair, poor appetite, hypersomnia, lowered activities , reduced food intake, loose stool, dark red tongue, white tongue with white, thick, greasy fur. Lep could be taken as one of objective indicators for evaluating hypertension rat model with EAPDS.

Animals ; Aorta ; Cell Proliferation ; Disease Models, Animal ; Hypertension ; physiopathology ; I-kappa B Proteins ; Interleukin-6 ; Leptin ; blood ; NF-KappaB Inhibitor alpha ; NF-kappa B ; Phosphatidylinositol 3-Kinases ; Rats ; Rats, Wistar ; Suppressor of Cytokine Signaling Proteins ; Transcription Factor RelA ; Tumor Necrosis Factor-alpha

Objective To explore the causes of subclinical hypothyrodism in children and the effects of the interventional therapy with thyroixine on the course of it.Methods Two hundreds children with subclinical hypothyroidism were measured for thyroglobulin antibody(TGAb),thyroid microsomal antibody(TMAb) in the blood serum,examined by colord Dopplor ultrasonic,examined by fine needle aspiraton cytology of the throid and measured the rate of 131I absorbed by thyroid in order to find out the causes of the disease.Two hundreds cases were randomly divided into two groups on the base of the cause of diseases,treatment group 100 cases and control group 100 cases.The treatment group were treated by throxine 25-75 ?g/d and the therapeutic dosage were chosen with the normal value of free triiodothyronine(FT3),free thyroxine(FT4)and high sensitive thyrotropin(sTSH) in the blood serum .After one year thyroxine therapy were stopped.Thyroid function was examined 6 months later after stopping the thyroxine.Results Among all of the causes of subclinical hypothyroidism in children,Hasgumoto′s thyroiditis accounts for 56%,simple goiter accounts for 26%,antithyroid drug accounts for 6%,the lack of thyroxine substitution therapy on the hypothyroidism accounts for 5% and undefined causes accounts for 7% .The thyroid function could keep normal for 1 year with an alternative therapy with thyroxine on subclinical hypothyroidism in children.Half a year later after stopping thyroxine,the thyroid function turned normal in most of the children.There were obvious differences in the ratio of cure and the ratio of effectiveness between treatment group and control group (t=20.2,3.2 Pa

BACKGROUND: Establishment of drug (neomycin or hygromycin) resistant feeder layer cells is necessary for screening the target gene positive clone of embryonic stem cell (ESC) transfected in vitro, and the establishment of drug-resistant NIH3T3 cell line is also important for the screening of other target ESC genes.OBJECTIVE: To obtain a feeder layer of positive cell clone of ESC transfected by pTet-on gene by means of the G418-resistant NIH3T3 cell line establishment.DESIGN: Cell culture and DNA examination.SETTING: Faculty of Laboratory Animal, China Medical University.MATERIALS: NIH3T3 cells were contributed by the Cell Biology Staff Room of China Medical University, and pWL/neo plasmid was a gift from Professor Jin Zhuang of Harvard University Medical College. G418,leukemia inhibitory factor (LIF, ESC GRO 106 U/mL) and DMEM were produced by GIBCO BRL Company, while mitomycin-C (MIT-C), DIG marking and kits were the products of Roche Company. Lipoetin was the product of Invitrogen Company, and bought from Shenyang Lianxing Biotechnology Co.,Ltd.METHODS: ①The pWL/neo plasmid containing neo gene was purified and transfected into NIH3T3 cells by lipoetin.②The transfected cells were further cultured in 25 mL medium containing G418 antibiotic of gradient concentration to observe the survival and apoptosis of cells and measure G418 minimal fatal dose (MFD) to NIH3T3 cells.③The transfected cells were subcultured, and the clone of single cell was selected to 24-pore culture board for screening amplification. At the same time, normal NIH3T3 cells were also selected and added with the selective culture medium at the same dose of G418, as screening negative control. ④The cells were planked with lower cell density, and further screened in DMEM medium containing G418 MFD; Meanwhile, normal NIH3T3 cells were taken as controls, and cultured by G418 contain MFD and by DMEM medium without G418 respectively. Light microscope was used to observe G418R NIH3T3 cells. ⑤G418R NIH3T3 cells and MEF were all applied as the feeder layer to culture the ES-D3 cells, which were observed by light microscope. Cell DNA was prepared, and evaluated by PCR and southern blot.MAIN OUTCOME MEASURES: ①G418 MFD to NIH3T3 cells. ②screening result of G418R NIH3T3 cells. ③growth of G418R NIH3T3 cells. ④growth of ES-D3 cells on G418R NIH3T3 cells.RESULTS: ①G418 MFD to NIH3T3 cells was defined as 500 mg/L.②Under the condition of G418 (500 mg/L), G418R NIH3T3 cell clones were successfully selected. ③The G418R NIH3T3 cells had no difference from normal NIH3T3 cells in morphology and propagation rate.④The ESC cultured in feeder layer present a colony growth, smooth limitation and remained an undifferentiation state. The resistant cell genomic DNA of G418R NIH3T3 cells was assayed with specific neo gene primer, and then neo gene DNA fragment was amplified; Southern blot analysis showed that neo gene fragment integrated into G418R NIH3T3 cells.CONCLUSION: The G418R NIH3T3 cells are established successfully,and the ESC cultured in the G418R NIH3T3 cell feeder layer can keep the characteristics of normal ESC.

OBJECTIVE: To observe the time-course expression of zonula occludens-1 (ZO-1) in cerebral cortex after traumatic brain injury (TBI). METHODS: The TBI model of mouse was established. The mice were divided in 1 h, 3 h, 6 h, 12 h, 24 h, 3 d, 7 d after TBI, sham and control groups. The permeability of the blood brain barrier was evaluated by measuring the extravasation of Evans blue (EB) dye. The expression of ZO-1 in cerebral cortex in the injured area was detected by Western blotting and immunohistochemistry. RESULTS: The extravasation of EB dye of injured cortex gradually increased from 1 h, peaked at 1-3 d and approximately decreased to normal at 7 d after TBI. Western blotting revealed that the expression of ZO-1 gradually decreased after 1 h, was at the lowest at 1-3 d, and then significantly increased after 7 d but was still lower than that of normal and sham groups. The result of immunohistochemistry showed that ZO-1 had strong expression in vessel of normal cortex, gradually decreased after TBI, and almost disappeared at 3 d after TBI and gradually recovered to normal level later. CONCLUSION The expression of ZO-1 in the injured cortex after TBI initially decreases and then increases. The negative correlation between ZO-1 expression and EB extravasation after TBI could be used as a new indicator for wound age estimation.
Animals ; Blood-Brain Barrier ; Blotting, Western ; Brain Injuries/physiopathology* ; Cerebral Cortex/metabolism* ; Immunohistochemistry ; Mice ; Permeability ; Tight Junctions/metabolism* ; Zonula Occludens-1 Protein/metabolism*

navirus disease 2019 (COVID-19) in the patients younger than 18 years old infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant, and to provide a basis for determining the chest CT changes and efficacy of COVID-19 caused by Omicron virus variant in patients younger than 18 years old. Methods　The clinical and imaging data of 30 cases of patients younger than 18 years old infected with COVID-19 Omicron variant, who admitted to the Third People's Hospital of Shenzhen from February 11 to March 26, 2022 were collected and retrospectively analyzed. The clinical manifestations, imaging features and dynamic changes of lesions were summarized. Results A total of 41 intrapulmonary lesions in 30 patients with COVID-19 caused by SARS-CoV-2 Omicron variant. The main manifestations were patchy or nodular ground-glass opacities and/or consolidation, with focal subpleural distribution, lesions mainly occur in the right lung (70.73%, 29/41). There were 42 lesion morphologies, with 22 (52.38%) striped shadows and 16 (38.10%) nodular shadows, with small lamellar and patchy shadows predominating. There were 36 lesion density variations, with ground glass shadows being the most common, with a total of 24 ground glass shadows (66.66%) in each lobe of the lung, and also 6 consolidation lesions (16.67%) and 6 mixed ground glass opacity and consolidation lesions (16.67%). With the progression of the disease, lesions gradually enlarged, appeared on the 2nd day (312.93 mm3), peaked on the 9th day (1 837.18 mm3). The average absorption time of the lesions was (16±3) days, and there was no significant difference between the absorption time of patchy and nodular lesions (ground glass and/or consolidation) (t=0.853, P>0.05). The lesions showed focal ground-glass opacity in the early stage, 77.78% lesions were absorbed after treatment in the late stage. Inflammatory nodules were absorbed slowly (9-19 days), without residual fibrotic changes. Conclusions　The imaging manifestations of COVID-19 in patients younger than 18 years old infected with SARS-CoV-2 Omicron variant have certain characteristics, showed patchy or nodular ground glass opacities and/or consolidation, mainly distributed in the subpleural area, with small and few lesions and slow change, didn't remain fibrosis. Being familiar with its clinical and imaging manifestations can assist in early diagnosis, but confirming the diagnosis requires a combination of epidemiological history, clinical symptoms, SARS-CoV-2 nucleic acid and radiological manifestations.

Macrolactins are 24-membered macrolides produced by unidentified marine bacterium, Actinomadura sp. and Bacillus sp., which exhibit both antibacterial and antitumor activities in vitro. The environmental strain X-2 which was isolated from the sediment of the East China Sea produce Macrolatin A, B and O. In this study, a set of degenerate oligonucleotide primers, designed for amplification ketosynthase(KS) domains, had been employed to identify KS gene fragments of the X-2 DNA samples. One 645 bp KS fragment(GenBank accession no. EF486351)had been cloned and used as a probe to screen the genome DNA fosmid library of X-2. Three positive clones were selected and sequenced, Homologous analysis and the function prediction of the obtained PKS gene fragments suggested that macrolactin is the Polyketide Biosynthesis Product of the gene cluster obtained in the environmental strain X-2.