Objective To explore the clinical manifestations and epidemiological profiles of hospitalized infants with Bordetella pertussis infection and to improve the knowledge for its early diagnosis and treatment.Methods Using a retrospective analysis method,the data of 142 hospitalized infants with Bordetella pertussis infection in Shenzhen Children's Hospital from 2011 to 2013 were collected,which was confirmed by a real-time polymerase chain reaction(PCR) of nasopharyngeal secretion.The SPSS 16.0 software was used for statistical analysis.Results Of the 142hospitalized infants with Bordetella pertussis infection,the ratio of male to female was 2.1∶ 1.0 (96/41 cases).Ninetythree (65.5％) cases were aged less than 3 months old,26 cases (18.3％) were 3-6 months old and 23 cases (16.2％) were 6-36 months old.One hundred and twenty-six (88.7％) patients were unvaccinated or incompletely vaccinated.The most common clinical manifestation was paroxysmal cough which accounts for 104/142 (73.2％) cases,and 6 cases of which had roaring(4.2％).Peripheral blood WBC level of 83 cases increased,and 75 (90.4％) cases were lymphocytosis.The positive rate of X-ray was 60.6％.Eighty-one cases were complicated with bacteria,respiratory virus or mycoplasma pneumonia infection.There were significant differences in symptom of fever and infection of respiratory tract pathogen among 3 groups.The symptom of fever in the group aged less than 3 months old group was significantly lower than the other 2 groups (x2 =11.45,P ＜ 0.01 ;x2 =34.56,P ＜ 0.01),and 6-36 months old group was significantly higher than the other 2 groups(x2 =7.590,P ＜ 0.01 ;x2 =6.928,P ＜ 0.01).Conclusions Bordetella pertussis is an important pathogen for the infants with persistent cough,especially in unvaccinated or incompletely vaccinated infants,but the clinical manifestation and signs of pertussis are atypical,which may lead to a misdiagnosis and delay proper treatment.As a rapid and sensitive method of detecting Bordetella pertussis,PCR may be used for early diagnosis.

Objective:To determine the difference of miRNA expression profiles in 6 types of human cancer cells by microarray technique.Methods:The microarray was prepared,with contained 210 oligonucleotides,including 206 probes complementary with human and mouse miRNA sequences and 4 positive control oligos.MiRNAs were extracted from HeLa(human cervical cancer epithelial cells),MCF-7(human breast cancer cells),A549(human lung adenocarcinoma cells),HT-29(human colonic cancer cells),ES-2(ovarian carcinoma cells),and K562(chronic myelogenous leukemia cells) cells and were labeled with Cy3 for hybridization to the miRNA microarray.The slides were scanned by ScanArrayTMExpress1.0 and images were analyzed by ScanArray3.0 and Cluster3.0;the results were confirmed by Northern blotting and RT-PCR.Results:Totally 115 miRNAs were found to be differentially expressed in the 6 cancer cell lines,with miR-21 expression up-regulated and miR-125b,let-7 expression down-regulated.The expression of miR-17-5p and miR-20a was in cluster and was more higher in ES-2 cells than in other cells.HeLa and MCF-7 cells were located on a single branch of the dendrogram in cluster analysis.Northern blotting showed that both pre-and miR-17-5p expressed in K562,pre-miR-17-5p was weakly expressed in A549 and ES-2 cells,and obvious pre-miRNA expression and weak miRNA expression were noticed in MCF-7,HeLa,and HT-29 cells.RT-PCR showed that expression of pre-miR-17-5p in K562 cells was higher than those in other cells.Expression of miR-21was high in all 6 cell lines and the highest expression was seen in A549 cells.Conclusion:Microarray can be used to detect miRNA expression files in cancer cells,which contributes to the study of the relation between miRNA and tumor.

This study explored the possibility that the components in melanoma cytoplasm induce murine BMSCs transformation and expression of Melan-A by morphologically observing the changes of BMSCs and immunocytochemically detecting Melan-A in the cells after culturing BMSCs in medium containing melanoma cytoplasm components (MCC). MCC of B16 melanoma cells was prepared and BMSCs were cultured and induced by adding the MCC into culture medium. The cells were morphologically observed and Melan-A was immunohistochemically detected to confirm BMSCs transformation. MCC-induced BMSCs underwent morphological changes. A number of melanin granules appeared in the cytoplasm of the cells and some were released into surrounding areas. Several cells that might come from one cell formed a cluster, and their granules, together with those secreted by other induced BMSCs, formed a so-called "sphere-formed structure". The induced BMSCs expressed Melan-A. We are led to conclude that there might be some factors in the cytoplasm of melanoma cells that might induce BMSCs transformation toward melanogenic cell, or even melanoma.

To evaluate the specific inhibition of antisense u-PAR on the u-PAR expressions in highly invasive cell subclones and to determine its blocking function in the invasion by those cells, a cDNA fragment of u-PAR obtained by RT-PCR was inserted into a plasmid vector named pcDNA3 in antisense orientation. Then the antisense u-PAR recombinant was transfected into highly invasive cell subclones. The u-PAR expression in neo-resistant cells was examined by RT-PCR and immunohistochemical assay. Compared to the control cells, the content of mRNA and protein of u-PAR in transfected cells decreased sharply, and the rate of inhibition was 53% and 73%, respectively, indicating that an antisense u-PAR might have played a specific inhibitory role in its expression in the cells, which may provide a good cell model for making further investigation of the inhibitory effects of the antisense u-PAR on invasion in highly invasive cell subclones of human prostate carcinoma.
Cell Line, Tumor ; Cloning, Molecular ; Humans ; Male ; Neoplasm Invasiveness ; Plasmids ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Antisense ; biosynthesis ; genetics ; Receptors, Cell Surface ; biosynthesis ; genetics ; Receptors, Urokinase Plasminogen Activator ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Urokinase-Type Plasminogen Activator ; antagonists & inhibitors ; genetics ; metabolism

Objective:To observe any effect of combining ultrasound-guided stellate ganglion pulsed radiofrequency irradiation with electroacupuncture in treating migraine.Methods:Seventy-two migraine patients were randomly divided into a combined group, an electroacupuncture group and a control group, each of 24. All three groups were treated with etocoxib and zolmitriptan dispersible tablets, while the electroacupuncture group and the combined group were additionally provided with electroacupuncture or ultrasound-guided stellate ganglion pulsed radiofrequency irradiation combined with electroacupuncture. The therapeutic interventions were administered once daily for 7 days. A visual analogue scale (VAS), a migraine-specific quality of life questionnaire (MSQOL), the Hamilton anxiety scale (HAMA), the Hamilton depression scale (HAMD) and a migraine disability scale (MIDAS) were used to evaluate each patient before the experiment and on the 3rd, 7th, 30th and 90th days after completion of the treatment.Results:The average VAS, MSQOL, HAMA and HAMD scores of the combined and acupuncture groups had improved significantly at all time points. Moreover, the average MIDAS scores of the combined and acupuncture groups had improved significantly 30 and 90 days after the treatment. On the 3rd day after the treatment, the average VAS, MSQoL, HAMA and HAMD scores of the combined group were significantly higher than the acupuncture group′s averages, while the average MIDAS score of the former group was significantly lower on the 30th and 90th days after the treatment. The average VAS, HAMA and HAMD scores of the combined group were significantly lower than the control group′s averages 3, 7, 30 and 90 days after the treatment, while their average MSQOL score was significantly higher. The average MIDAS score of the combined group was significantly lower than the control group′s average 30 and 90 days after the treatment.Conclusion:Ultrasound-guided stellate ganglion pulsed radiofrequency irradiation combined with electroacupuncture can significantly relieve the symptoms of migraine and improve the life quality of migraine patients for at least 3 months.

Objective:To observe the clinical efficacy of combining repeated transcranial magnetic stimulation (rTMS) with radiofrequency ablation (RF) of dorsal root ganglia in treating herpes zoster infection and neuralgia.Methods:Eighty-four individuals with a herpes zoster infection who had suffered from neuralgia for no more than 7 days were divided randomly into a control group, an rTMS group, an RF group, and an observation group, each of 21. All were treated with gabapentin, valciclovir and mecobalamin. The rTMS group received rTMS treatment, 5 days per week, for 2 consecutive weeks. The RF group received RF treatment of the dorsal root ganglia on the 15th day after enrollment, while the observation group received only the medication. Before the experiment as well as after 3, 7, 16, 30 and 60 days, all of the subjects self-assessed their discomfort using a pain visual analog scale (VAS). The Hamilton Depression Scale (HAMD), Hamilton Anxiety Scale (HAMA), Quality of Life Assessment Scale (QOL-SF36), and Pittsburgh Sleep Quality Index (PSQI) were also administered.Results:The average VAS, HAMA, HAMD, QOL-SF36, and PSQI scores of the observation group improved continuously and significantly during and after the treatment. Beyond 16 days all of those results were significantly better than the control group′s averages, and the observation group′s average VAS, HAMA and HAMD results were also significantly better than those of the rTMS group. The observation group′s average VAS, HAMA, HAMD and PSQI scores had improved significantly more than the RF group′s averages beyond 30 days.Conclusion:Combining rTMS and dorsal root ganglion RF can effectively alleviate the early pain symptoms of herpes zoster infection and neuralgia, relieve anxiety and depression, and significantly improve sleep and life quality. Such therapy is worthy of clinical promotion and application.

This study explored the possibility that the components in melanoma cytoplasm induce murine BMSCs transformation and expression of Melan-A by morphologically observing the changes of BMSCs and immunocytochemically detecting Melan-A in the cells after culturing BMSCs in medium containing melanoma cytoplasm components (MCC).MCC of B16 melanoma cells was prepared and BMSCs were cultured and induced by adding the MCC into culture medium.The cells were morphologically observed and Melan-A was immunohistochemically detected to confirm BMSCs transformation.MCC-induced BMSCs underwent morphological changes.A number of melanin granules appeared in the cytoplasm of the cells and some were released into surrounding areas.Several cells that might come from one cell formed a cluster,and their granules,together with those secreted by other induced BMSCs,formed a so-called “sphere-formed structure”.The induced BMSCs expressed Melan-A.We are led to conclude that there might be some factors in the cytoplasm of melanoma cells that might induce BMSCs transformation toward melanogenic cell,or even melanoma.

To observe the inhibitory effects of an antisense u-PAR vector on invasion of highly invasive PC-3M cell subclones, the effects of the antisense u-PAR on activity of MMP-9 in those highly invasive cell subclones were detected by a quantitative RT-PCR and zymography. The monolayer invasion assay and colony formation assay in soft agar were used. And tumorigenesis rate and invasions by the cell subclones with or without the antisense u-PAR were observed in nude mice. It was found that in vitro growth of highly invasive PC-3M cell subclones transfected with the antisense u-PAR was declined, and the ability of anchorage-independent growth of those cell subclones was found decreased sharply, with the inhibiting rate becoming 79% and 60%, respectively. Although the antisense u-PAR didn't change MMP-9 gene transcription, they could inhibit the activation of MMP-9 of highly invasive PC-3M cell subclones. Moreover, the tumorigenesis rate of the cell subclones with the antisense u-PAR decreased and the growth of a neoplasm also slowed down. The t tests showed the difference between experimental and control groups was statistically significant (P < 0.01). The antisense u-PAR vector could not only inhibit the invasion ability of highly invasive PC-3M cell subclones in vitro but also restrain the growth of those cell subclones in vivo.
Animals ; Antisense Elements (Genetics) ; genetics ; pharmacology ; Cell Division ; drug effects ; Cloning, Molecular ; Humans ; Male ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Antisense ; Receptors, Cell Surface ; genetics ; metabolism ; Receptors, Urokinase Plasminogen Activator ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Tumor Cells, Cultured

OBJECTIVE：To optimize the water extraction and ethanol precipitation technology of Zhuang medicine Baijin granules. METHODS ：HPLC method was adopted to determine the content of bergenin in Baijin granules extract. The extraction routes of Baijin granules （water decoction ，70％ ethanol reflux extraction ，water decoction combined with 70％ ethanol reflux extraction）was screened primarily with the yield of extract and the experiment of reducing uric acid of mice. The orthogonal test was adopted to optimize water extraction technology of Baijin granules with water multiple ，extraction time and extraction times as factors，taking the extraction yield and the bergenin content as index ，then the validation test was carried out. The orthogonal test was adopted to optimize the alcohol precipitation process of Baijin granules including the relative density of medicinal materials ， alcohol content and the alcohol precipitation time ，then the validation test was carried out. By the experiment of reducing uric acid ， the effects of medicinal materials extract of Baijin granules extract were compared before and after ethanol precipitation. RESLUTS：Established method for content determination of bergenin with linearity range of 0.007 2-0.288 mg/mL，had good precision，reproducibility，stability and accuracy. The initially chosen extraction process of Baijin granules was water decoction extraction. The optimal water extraction technology was soaked for 0.5 h，then decocted for 3 times with 14-fold water （mL/g）and 1.0 h each time. The optimal ethanol precipitation process was to concentrate the water extract to a relative density of 1.0 g/mL with alcohol content of liquid at 60％ and precipitated for 12 h. Validation tests showed that RSDs of extract yield and beragenin content were all lower than 2％（n＝3）. The experiment of pharmacodynamics showed that water extract （before ethanol precipitation ）and water extract after alcohol precipitation could significantly decrease the level of uric acid in hyperuricemia model mice （P＜0.01）. There was no statistical significance in the reduction of uric acid between 2 groups（P＞0.05）. CONCLUSIONS ：The optimized water extraction technology can obtain good extract yield and bergenin content ，and combined with ethanol precipitation technology for removing excess impurities would not affect the pharmacodynamics. The water extraction and ethanol precipitation technology is feasible，and can be used for extracting the medicinal materials of Baijin granules and its edulcoration.

OBJECTIVE： To provide reference for perfecting the quality standard revision of TCM preparations in medical institutions， and improving the quality standard of TCM preparations in medical institutions. METHODS： By sorting out the quality standards of TCM preparations in Guangdong provincial medical institution preparation specification， this paper summarized the main problems and put forward suggestions for improvement. RESULTS & CONCLUSIONS： There are 897 kinds of TCM preparations included in the Guangdong provincial medical institution preparation specification. The recent quality standards （2017 edition） have been greatly improved compared with those of the first edition （1985 edition）； however， there are still some problems in the overall quality control of preparations， quantitative control of indicative components， project specificity， source control of medicinal materials， control of medicinal materials not included in the Chinese Pharmacopoeia and quality standards of the same series of varieties. It is suggested that on the premise of considering both advancement and applicability， the quality control of the whole， local and toxic preparations of TCM hospitals should be strengthened， the detection methods with good specificity and reproducibility should be properly updated， the control of key parameters of production process should be strengthened， the quality standard of TCM preparations in medical institutions should be fully improved， so as to provide the products with safe， effective and controllable in quality products in medical institutions.