Establishment of HPLC fingerprint and content determination of 8 components in Jianpi huayu decoction / 中国药房

OBJECTIVE To establish the HPLC fingerprint of Jianpi huayu decoction， and to determine the contents of 8 components. METHODS Thermo Hypersil Gold C18 column was used with mobile phase consisted of methanol-0.05% phosphoric acid aqueous solution （gradient elution） at the flow rate of 1.0 mL/min. The column temperature was 30 ℃， the injection volume was 5 μL. The detection wavelength of matrine was 211 nm， and the other components’ detection wavelength was 283 nm. The similarity evaluation of HPLC fingerprints for 10 batches of Jianpi huayu decoction was performed by using the Similarity Evaluation System of Chromatographic Fingerprint of Traditional Chinese Medicine （2012 edition）. The contents of chlorogenic acid， vanillic acid， p-coumaric acid， ferulic acid， hesperidin， quercetin， bergapten and matrine in the samples were determined by HPLC. RESULTS HPLC fingerprint of Jianpi huayu decoction was established. A total of 27 common peaks were identified， and 8 components were identified. The similarity between 10 batches of samples and the control map ranged from 0.942-0.999. RSDs of precision， repeatability and stability tests were less than 3% （n＝6）. The average recoveries of chlorogenic acid， vanillic acid， p-coumaric acid， ferulic acid， hesperidin， quercetin， bergapten and matrine were 99.48%， 101.32%， 101.18%， 100.79%， 101.12%， 99.19%， 99.81% and 102.46%， respectively； RSDs were 1.34%， 0.93%， 1.90%， 1.84%， 0.54%， 1.53%， 1.33% and 1.01%， respectively （n＝6）. The contents were 0.021-0.061， 0.025-0.034， 0.116-0.295， 0.006- 0.062， 0.014-0.053， 0.017-0.026， 0.014-0.027 and 14.05-24.11 mg/g， respectively. CONCLUSIONS The established fingerprint and content determination method can provide a reference for the quality control and subsequent preparation development for Jianpi huayu decoction.

Exploratory research of the value of abbreviated comprehensive geriatric assessment in elderly breast cancer patients / 中华老年医学杂志

Objective:To explore the value and feasibility of abbreviated comprehensive geriatric assessment(aCGA)grading in elderly patients with breast cancer.Methods:From June 2019 to January 2020, elderly patients with breast cancer aged 65 years and above were enrolled.Eastern Cooperative Oncology Group(ECOG)score and aCGA classification were performed respectively.Clinical characteristics, score distribution and differences between the two assessment methods were compared and analyzed.Results:A total of 61 cases of breast cancer patients aged 65 years and above were included in our study.According to the assessment of aCGA, grade A accounted for 65.5%(40/61), grade B accounted for 27.9%(17/61), and grade C accounted for 6.6%(4/61), among which 82.0%(50/61 cases)of the patients had complications.And the most common complications were hypertension, cardiovascular disease and diabetes.Among the 50 patients with ECOG score 0-1, 74.0%(37/50)were aCGA grade A, and 26.0%(13/50)were aCGA grade B.Conclusions:According to the aCGA grading, about two thirds of breast cancer patients over 65 years old are assessed as grade A, which indicates that they might have better tolerance during the treatment.However, among the patients with 0-1 score according to the ECOG score, some patients still have a slightly worse grade(aCGA grade B, which shows slightly worse health condition), suggesting that the refinement degree of ECOG score may be insufficient, and the health damage of some patients may be underestimated.

Mechanism of action of circular RNAs in hepatocellular carcinoma / 临床肝胆病杂志

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related death worldwide and has high mortality and recurrence rates and low survival rates. Circular RNAs (circRNAs) are a class of single-stranded closed circular RNA molecules with tissue- and development-specific expression and play a regulatory role in organisms. These circRNAs regulate the progression of HCC by acting as miRNA sponges and targeted protein-coding genes, but the basic mechanism is still unclear. This article summarizes the formation mechanism and biological functions of circRNAs, reviews the research advances in the role of circRNAs in the pathogenesis of HCC, and points out their potential value as diagnostic markers and therapeutic targets for HCC.

Patterns and practice management of occupational exposure of medical staffs in coronavirus disease 2019 isolation wards / 中华传染病杂志

Objective:To explore the patterns and causes of occupational exposure to infectious diseases (OEID) among frontline medical staffs (FMS) in coronavirus disease 2019 (COVID-19) isolation wards (CIW), and the particularity of post-OEID management and the measures to prevent OEID.Methods:A total of 1 061 FMS of Wuhan Huoshenshan Hospital from February 4 to March 21, 2020 were enrolled. The OEID of FMS was investigated and analyzed from the perspectives of FMS physical and psychological conditions, protective equipment, infection-control related regulations and procedures, local air quality, exposure patterns, and the particularity of emergency treatment after exposure.Results:The incidence of OEID among FMS was 2.0%(21/1 061). The nurses and doctors accounted for 95.2%(20/21) and 4.8%(1/21), respectively. The incidences in 17 general wards and two intensive care units (ICU) were 71.4%(15/21) and 28.6%(6/21), respectively. Nearly 90.5%(19/21) and 9.5%(2/21) of the OEID events occurred in contaminated area and potential contaminated area, respectively. About 23.8%(5/21) of the OEID events were air exposure of oral-nasal skin, mucosa and respiratory tract, which was secondary to uncontrollable vomiting, and 76.2%(16/21) were pricking injuries. The inducement factors involved poor quality and inappropriate wearing of some goggles, atomization of the inside of goggles leading to blurring vision, chest distress and decreased sense of touch and operational flexibility related to level-3 protection equipment, poor air quality, FMS physical and psychological conditions, etc. Under the direction of "the Procedures for Handling OEID" , all incidents are properly handled and no FMS was infected by 2019 novel coronavirus and blood-borne pathogens. No new OEID event was found after the strict implement of set of preventive measures.Conclusions:The OEID among FMS in CIW is attributed to multiple causes. The optimized process that takes into account the specificity of OEID management for both COVID-19 and blood-borne infectious diseases can effectively prevent potential post-exposure infections. And reasonable precautions can fully reduce the risk of OEID of FMS in CIW.

Comparative analysis of the correlation between axillary treatment and upper limb lymphedema / 中华外科杂志

In the surgical treatment of breast cancer, "de-escalating" is becoming more popular, especially in the treatment of axilla. Axillary sentinel lymph node biopsy has become a routine treatment for axillary-negative breast cancer patients because it can effectively reduce the occurrence of upper limb lymphedema, so that some patients with sentinel lymph node negative can be exempted from axillary lymph node dissection. However, in recent years, several international clinical studies, such as AMAROS trial, explored the use of radiotherapy instead of dissection when 1 to 2 sentinel lymph nodes were positive. The results showed that radiotherapy can reduce the incidence of upper limb edema by nearly 50% compared with dissection. How to interpret the results of such non-inferiority studies, and how to treat the impact of axillary dissection and axillary radiotherapy on the occurrence of lymphedema? Based on the review of literature in recent 10 years, the incidence and hazard ratio of edema related to the two axillary treatments were compared. The conclusion is the hazard ratios are close to each other, around 3 for both arms. The results of clinical studies such as radiotherapy instead of axillary dissection need to be carefully interpreted. At the same time, axillary dissection is still suitable for some patients at current stage, and the corresponding prevention of upper limb lymphedema still needs attention.

Advances in the role of exosome-derived long non-coding RNAs in hepatocellular carcinoma / 中华肝脏病杂志

Exosomes are one kind of microvesicles secreted by living cells. It contains some small non-coding RNAs (microRNA, lncRNA, circRNA, etc.) which play an important role in signal transduction. In addition, they transfer information through circulating small vesicles, regulate epithelial-mesenchymal transition, tumor microenvironment, promote microangiogenesis, regulate tumor immunity, chemoradiation resistance, and regulate cellular energy metabolism pathways, thereby promoting the occurrence, development and metastasis of hepatocellular carcinoma. This article reviews the mechanism of lncRNA in exogenous and in vivo in relation to the occurrence, development and metastasis of hepatocellular carcinoma.

Impact of the 21-gene recurrence score assay in clinical treatment and prognosis analysis for patients with hormone receptor positive early-stage breast cancer / 中华肿瘤杂志

Objective@#To explore the association between the 21-gene recurrence score (RS) and clinicopathologic characteristics as well as prognosis in patients with axillary lymph node negative, hormone receptor (HR) positive breast cancer.@*Methods@#The clinicopathologic data of 439 early breast cancer patients who underwent 21 gene RS testing was retrospectively analyzed. According to the 21 gene RS, the patients were divided into low risk (295 cases), intermediate risk (111 cases) and high-risk (33 cases) group. The relationship between the 21 gene RS and clinicopathological characteristics, treatment, recurrence and metastasis was analyzed. Univariate and multivariate statistical analyses were used to analyze the risk factors for relapse free survival (RFS).@*Results@#Tumor grade, estrogen receptor (ER), progesterone receptor (PR) and Ki-67 index were significantly different among the 3 risk cohorts (P<0.001 for all). After a median follow-up of 32 months, the recurrence rate in low risk group (3.7%) was significantly lower than that in the intermediate-high risk group (9.0%), the locoregional recurrence (LRR) rate of low, intermediate and high risk group was 2.4%, 6.3% and 9.1%; and the distant metastasis (DM) rate in low risk group was 1.4% and 2.1% in the intermediate-high risk group. Univariate analysis showed RS, ER status and endocrine therapy were prognostic factors for RFS (P<0.05 for all). Multivariate analysis showed that RS was an independent significant predictor for RFS (P=0.04).@*Conclusions@#The 21-gene RS is related to tumor grade, ER, PR and Ki-67 index. RS is an independent risk factor for RFS in patients with hormone receptor positive early-stage breast cancer.

Analysis of single circumaereolar incision nipple-sparing modified radical mastectomy and immediate tissue expander implantation / 中华内分泌外科杂志

Objective To explore the safety and cosmetic effect of nipple-sparing modified radical mas tectomy and immediate tissue expander implantation with single circumaereolar incision.Methods 30 patients were enrolled in Peking Union Medical College Hospital between Jan.2014 and Dec.2015.All the patients were categorized according to surgical incision (single circumaereolar incision group vs double incisions group).Data on clinicopathological parameters,average hospital stay,complications and overall cosmetic effect were retrospectively collected.Data was performed with Chi-square test,Fisher exact test and t-test.Statistical significance was defined as P＜0.05.Results 19 patients were enrolled in single circumaereolar incision group,and 11 patients in double incisions group.There was no significant difference for operation duration (P=0.093) and average hospital stay (P=0.339).After follow-up for 19.1 months,ranging from 8 to 31 months,no patients developed seroma or arm lymphedema.There was no statistical significance between the two groups in terms of sensation in nippleaereolar area (P=0.973),bilateral symmetry (P=0.650) and overall cosmesis (P=0.483).Conclusion single circumaereolar incision nipple-sparing modified radical mastectomy and immediate tissue expander implantation can be one of the preferable surgical procedures with benefits of minimal invasiveness,reliable oncological safety and decent cosmetic effect.

Focusing on the role of conjunctiva goblet cell in maintaining the health of ocular surface / 中华实验眼科杂志

Ocular surface tissues are vulnerable to various stimuli from the external environment,including microbial invasion,mechanical damage,chemical stimulation,dust,UV light,and allergen stimulation.Conjunctival goblet cells play an important role in maintaining ocular surface self-homeostasis and health.These cells are the only unicellular glands in humans and mammals that are mainly distributed in the epithelium of the digestive tract,respiratory tract,and the ocular surface.Many factors affect goblet cell development and differentiation,especially the newly discovered molecule Spdef,which is a transcription factor found in the ETS family.The main function of conjunctival goblet cells is to secrete mucin to protect and lubricate the ocular surface;however,they also secrete a variety of other bioactive substances.Recent studies have demonstrated that goblet cells are involved in antigen presentation,regulation of dendritic cell phenotype differentiation,and induction of immune tolerance on the mucosa.Many ocular surface diseases,such as dry eye,will alter density and function of conjunctival goblet cells.Therefore,ophthalmologists should be concerned about the biological behavior of conjunctival goblet cells and the relationship between them and ocular surface diseases.

Purification and enzyme activity assay of filamentous temperature-sensitive protein Z in Streptococcus suis serotype 2 / 中国人兽共患病学报

We conducted purification of filamentous temperature-sensitive protein Z of Streptococcus suis serotype 2 (S.suis 2) and measured its GTPase activity.The ftsz gene in the genome of the Chinese 05ZYH33 strain of S.suis 2 was successfully amplified using PCR,and then the ftsz gene was cloned into prokaryotic expression plasmid pET28a,and the recombinant plasmid pET28a-ftsz was transformed into E.coli BL21.After induction by IPTG,the isolated FtsZ protein was analyzed with SDS-PAGE.Then the recombinant protein was purified by Ni2+-NTA affinity chromatography.The rabbit serum was harvested after immunization with recombinant FtsZ protein,and was analyzed by indirect ELISA and Western blotting.The GTPase activity of FtsZ was measured with the malachite green method.Results showed that successfully constructed recombinant plasmid pET28a-ftsz and the recombinant protein with high purity was obtained;Western blot result indicated that FtsZ could react with the His-tag antibody and the rabbit serum;the polyclonal antibody titer of the rabbit serum reached 1 ∶ 13 107 200;FtsZ have GTPase activity.We successfully prepared S.suis 2 recombinant protein FtsZ having GTPase activity and high titer antiserum would be useful for the further study of S.suis 2 cell division mechanism.

Trastuzumab administered concurrently with anthracycline-containing adjuvant regimen for breast cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the safety and efficacy of trastuzumab administered concurrently with anthracycline-containing adjuvant regimen for breast cancer.</p><p><b>METHODS</b>It is a prospective, randomized and controlled trial. Participants were randomized to receive trastuzumab administered concurrently or sequentially with anthracycline-containing adjuvant regimen. The primary endpoint was cardiac safety. The second endpoints were disease-free survival (DFS) and overall survival (OS).</p><p><b>RESULTS</b>One hundred and nine breast cancer patients were enrolled and randomized in this trial. Fifty-five participants received trastuzumab administered concurrently with anthracycline-containing adjuvant regimen and 54 patients received trastuzumab administered sequentially with anthracycline. The primary cardiac event was asymptomatic decrease in the left ventricular ejection fraction (LVEF). There was no significant difference between concurrent and sequential groups in cardiac event rates (9.1% vs13.0%, P = 0.556), neither of LVEF values at basline or at 3, 6, 9 and 12 months during trastuzumab treatment (P > 0.05). Four patients (7.3%) in the concurrent group suffered local recurrences or distant metastases, and 6 participants (11.1%) in the sequential group had distant metastases. There was no significant difference between the two groups in DFS (P = 0.724). There was no death in both groups.</p><p><b>CONCLUSIONS</b>Trastuzumab administered concurrently with anthracycline is a safe adjuvant regimen for breast cancer and does not increase cardiac events. Further research is needed to determine the efficacy of this treatment regimen.</p>

ERβ expression in breast cancer at different stages and molecular subtypes / 中华普通外科杂志

Objective To explore the association of estrogen receptor β expression with different stages and molecular subtypes of invasive breast cancer.Methods The clinicopathologic data of 446 invasive breast cancer cases was retrospectively analyzed.ERβ expression was evaluated by types and stages.Results Of all 446 invasive breast cancer cases,328 (73.5％) were ERβ positive.The ERβ positive rate was 77.9％ (240/308) and 63.8％ (88/138) in ERα + group and ERα-group,respectively.The ERβ expression in breast cancer was positively correlated with ERα (P ＜ 0.01) while it had no correlation with PR,histological grade,HER-2 and Ki-67 (P ＞ 0.05).ERβ expression was not significantly different among different age,tumor size and axillary lymph node groups(all P ＞ 0.05).A total of 418 invasive breast cancer cases were recruited for pathologic stage and NPI analysis,including 168 cases at stage Ⅰ,152 cases at stage Ⅱ and 98 cases at stage Ⅲ.ERβ expression was not significantly different among different stages of breast cancer(P =0.743).Analyzed in these 418 cases,NPI was ＜ 3.4 in 126 cases,3.4-5.4 in 207 cases and ＞ 5.4 in 85 cases.ERβ expression was not significantly different among different NPI group (P =0.644).The ERβ positive rate in Luminal A subtype,Luminal B1 subtype,Luminal B2 subtype,HER-2 subtype and TN subtype was 75.6％ (88/118),75.9％ (110/145),85.2％ (46/54),68.4％ (39/57) and 62.5％ (45/72) respectively.ERβ expression was significantly different between Luminal subtype and non-Luminal subtype (P =0.007).Conclusions ERβ was not differentially expressed among different breast cancer stages and NPI groups.ERβ was differentially expressed in different breast cancer molecular subtypes.

Advances in breast cancer related nomograms / 中国癌症杂志

Breast cancer is the leading cause of malignancy-related mortality in women worldwide. The more accurate prediction of lymph node metastasis and evaluation of personalized prognosis of breast cancer patients could provide evidence and reference for individualized comprehensive treatment and clinical decision-making. Nomogram is statistical calculation model developed to generate individualized prediction of a certain clinical event through the factors associated with it. Currently breast cancer related nomogram models is most commonly used in the prediction of non-sentinel lymph node status in patients with sentinel lymph node-positive breast cancer, sentinel lymph node metastasis in clinical node-negative breast cancer and prognosis evaluation of breast cancer. This article reviewed the recent advances in breast cancer related nomograms according to the above mentioned three aspects, and evaluated respectively the predictive factors, accuracy, characteristics and clinical application potential.

Expression of human Streptococcus suis 2 muramidase released protein and preparation and application of its monoclonal antibodies / 中国兽医学报

The mrp gene of SS2 human strain Habb was truncated and cloned into a prokaryotic expression vector pGEX4T-2,and a fusion-expressed protein MRP-GST of 61 000 was obtained in E.coli.The GST was cut from MRP-GST with thrombin protease to gain the purified MRP of 35 000 which showed a strong reaction to the SS2 positive sera in Western blotting.BALB/c mice were immunitied intraperitoneally with purified MRP protein.Murine myeloma cells were fused with the splenocytes of the immunized mice after the third immunization.An indirect ELISA coated with purified MRP was used to screen hybridomas for production of specific antibody.The six McAb recognized MRP specially.According to the results of 71 standard strains (48 SS and 34 SS2)by Sandwich ELISA,the coincidence rate was 97.2%,but for 34 SS standard serotype was 100 %.The ELISA method has a potential value for clinical and epidemiological applicationgs.

Investigation of the epidemiological and clinical characteristics of an outbreak of tsutsugamushi disease in Anhui Province / 中华传染病杂志

Objective To investigate the epidemiological and clinical characteristics of an outbreak of tsutsugamushi disease in Chuzhou region, Anhui Province, and to clarify the new changes of epidemic focus of tsutsugamushi disease in China. Methods Field epidemiological investigation and analysis of clinical features were done. The detections of specific antibodies against Rickettsia tsutsugamushi were conducted to diagnose tsutsugamushi disease using colloidal gold immunochromatography assay combined with Well-Felix reaction. The geomorphic and climatic characteristics of the new epidemic focus were investigated. Results The outbreak occurred from October to November, 2007. The epidemic focus located on mountainous brushland regions, and the air temperature fluctuated from 20-4 ℃. Nineteen cases of tsutsugamushi disease in the new-found epidemic focus were finally diagnosed, 9 cases out of them were hospitalized, another 9 had recovered when diagnosed by serological tests; the remaining one had classical manifestations of tsutsugamushi disease but did not receive the serological test for certain cause. The main clinical symptoms were chilly in 14 cases, fever in 19 cases, headache in 15 cases; among the 9 hospitalized patients, the symptoms were lymphadenectasis in 8 inpatients, skin rash in 7 inpatients, splenomegaly in 4 inpatients and skin eschar and ulcer in 7 inpatients and Weil-Felix reaction by OXκantigen positive in 4 cases; the specific antibodies against Rickettsia tsutsugarnushi of 18 tested cases were all positive. No severe complications occurred in all patients. Before the first case was identified, all other cases were not diagnosed in time and did not receive correct antibiotic treatment. Nine hospitalized patients recovered rapidly with the treatment of doxycycline. Conclusions The outbreak of tsutsugamushi disease in Anhui Province in 2007 is type of emerged in autumn and transitional epidemic focus. There is epidemic focus of tsutsugamushi disease in northern region of Anhui Province. Doxycycline is rapid and effective for the treatment of tsutsugamushi disease.

Si-Jun-Zi decoction repairs mitochondrial damage of cells of liver,myocardium, gastric mucosa and skeletal muscle in rats with spleen asthenia / 中国组织工程研究

BACKGROUND: Spleen is correlated with mitochondrion. The "spleen governs movement and transformation of food and liquids" in traditional Chinese medicine refers to not only the digestion and absorption of food in gastrointestinal tract, but also the process of biological oxidation and energy production of mitochondrion.OBJECTIVE: To analyze the effects of Si-Jun-Zi decoction (SJZD), a typical prescription for invigorating spleen and replenishing qi, on repairing the mitochondrial damage of cells of liver, myocardium, gastric mucosa and skeletal muscle in rats with spleen asthenia.DESIGN: A randomized controlled observation.SETTINGS: Second Department of Internal Medicine, the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine;Guangzhou University of Traditional Chinese Medicine.MATERIALS: The experiments were carried out in the internal medicine laboratory, the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine and the testing center of Guangzhou University of Traditional Chinese Medicine from June to December in 2004. Forty SD rats were provided by the animal center of Guangzhou University of Traditional Chinese Medicine. Xiao Cheng-Qi decoction consisted of officinal magnolia bark, immature bitter orange and rhubarb according to the ratio of 3:3:2; SJZD consisted of radix codonopsitis pilosulae, largehead atractylodes rhizome, India bread and liquorice root according to the ratio of 2:2:2:1, and it was provided by the Department of Pharmacy, the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,and made into 100% decoction.METHODS: After raised for 1 week, the SD rats were divided randomlyinto 4 groups: normal control group, spleen asthenia group, natural convalescence group and SJZD-treated group, and the rats in the latter three groups were made into models of long-term spleen asthenia. Rats in the normal control group were fed with normal chow, intragastric administered with saline (3 mL), once every other day for 34 weeks. Rats in the spleen asthenia group were intragastricly administered with Xiao Cheng-Qi decoction (3 mL) and fed every other day for 34 weeks, the rats in the natural convalescence group were fed with normal chow for 8 weeks after being treated as those in the spleen asthenia group for 26 weeks, and those in the SJZD-treated group were treated as those in the spleen asthenia group for 26 weeks, and then intragastricly administered with SJZD (4 mL, once a day) and fed with normal chow for 8 weeks. At the end of the 34th week,the rats were decapitated under anesthesia, and the skeletal muscle, liver,gastric mucosa and myocardium were taken out rapidly. The protein amounts in mitochondrial suspension were detected with the biuret method,the content of mitochondria in I g tissue was calculated according to the tissue mass, and the mitochondrial ultrastructures were observed under transmission electron microscope.MAIN OUTCOME MEASURES: The contents and ultrastructures of mitochondria in skeletal muscle, liver, gastric mucosa and myocardium were observed.RESULTS: All the 40 rats were involved in the analysis of results without deletion. ① At the end of the 34th week, the mitochondrial contents of skeletal muscle, liver, myocardium and gastric mucosa were all significantly lower in the spleen asthenia group than in the normal control group (P＜ 0.01), and markedly higher in the natural convalescence group than in the spleen asthenia group (P ＜ 0.05-0.01). The mitochondrial contents of the tissues were the highest in the SJZD-treated group, which were significantly higher than those in the spleen asthenia group and natural convalescence group (P ＜ 0.05-0.01), as well as the normal control group (P＜ 0.01). ② Comparison of mitochondrial ultrastructures in skeletal muscle,liver, gastric mucosa and myocardium at the end of the 34th week: In the spleen asthenia group, the mitochondria of myocardial cells were seriously swollen, the compact substance of hepatocellular mitochondria were decreased or disappeared and the crest disrupted; the mitochondria of skeletal muscle were shrunk and decreased, mitochondrial membranes were disorganized and crest disappeared, mitochondrial membranes were disorganized and crest disappeared; For gastric parietal cell of spleen asthenia,the amount of mitochondria reduced, inner structure confused, mitochondrial crestae of gastric chief cell was broken. In the natural convalescence group, the changes of the mitochondrial morphology were slight. The mitochondrial morphology in the SJZD group was close to those in the normal control group.CONCLUSION: SJZD has the effects of increasing the contents of mitochondria in skeletal muscle, liver, myocardium and gastric mucosa and repairing the damaged structure of mitochondria because of spleen asthenia.

Effects of superparamagnetic iron oxide on proliferation and neural differentiation of human mesenchymal stem cells / 基础医学与临床

Objective To label Flk-1+CD31-CD34-human mesenchymal stem cells (hMSCs) with superparamagnetic iron oxide (SPIO) and to evaluate the effect of SPIO on proliferation and neural differentiation of labeled cells. Methods hMSCs were incubated with SPIO (50 mg/L) and PLL (1.5 mg/L) overnight(12～18 hours). Both labeled and unlabeled cells went through growth curve test,Trypan blue staining and flow cytometer to evaluate the effects of SPIO on cell proliferation,cell viability and surface markers. Immunofluorescence assay was conducted for neuron and neuroglia specific cell surface markers after neural induction protocols were used. Results Cell viability of the two groups were both more than 90% for 7 days. There was no significant difference in cell viability and growth curve test between two groups. The results of flow cytometer showed that both labeled and unlabeled cells expressed CD44, CD105 and Flk-1 markers, while CD31 and CD34 were negative. After neural induction, the statistical analysis of A value for all the markers showed no significant difference between the two groups.Conclusion SPIO, as MRI cellular contrast, is safe and efficient.

Effect of recombinant adenoviruses with CD/TK fusion suicide gene on human hepatocellular carcinoma cells / 中西医结合学报

OBJECTIVE: To investigate gene-therapy for human hepatocellular carcinoma with adenovirus vectors by double suicide gene CD/TK. METHODS: Double suicide gene CD/TK was liberated from eukaryotic vectors pCEA-CD/TK and subcloned into shuttle vectors, and the transfer plasmid pAdtrack-CMV-CD/TK was formed after linearizing with Pac 1. It was recombinated with pAdeasy-1 in bacteria BJ5183. The identified adenovirus plasmid was digested by Pac1 and was transfected into 293 cells to pack the adenoviruses. After PCR determination, its titre was measured, and the infection rate and efficacy were tested in human hepatocellular carcinoma cells. RESULTS: pAdtrack-CMV-CD/TK and pAd-CD/TK were tested by endonuclease digestion. Ad-CD/TK was produced in 293 cells, and the human hepatocellular carcinoma cells (SMMC7721) infected by Ad-CD/TK were killed after 5-FC was used, and bystander effects were observed. CONCLUSION: Recombinant adenoviruses with CD/TK fusion suicide gene have a high infection rate and efficacy for human hepatocellular carcinoma cells.

Establishment of Epstein-Barr virus-transformed B cell lines from healthy volunteers of type B / 医学研究生学报

Objective: To develop Epstein-Barr virus(EBV)-transformed human peripheral blood B cell lines from healthy volunteers of type B.Methods: B lymphocytes from healthy volunteers of type B capable of producing anti-A antibodies were transformed by EBV,while Cyclosporine A(CsA),as an immunosuppressive agent that could selectively inhibit T-lymphocytes and protect B-lymphocytes from regression,was used in the experiment.Then,the supernanant of the cell culture medium was tested with red blood cells of type A,B and O by agglutinin assay.Results: Twelve of the15 EBV-transformed B lymphocyte cell lines were established,with a success rate of 80%,while 9 human B lymphocyte lines secreted anti-A antibodies.Conclusion: Human B lymphocyte lines secreting antibodies to A antigens were successfully developed,which helps further studies of human blood specific monoclonal antibodies.

Cloning and expression of Escherichia coli O157∶H7 Stx2B subunit / 医学研究生学报

Objective: To clone,identify and express the Shigalike toxin 2B(Stx2B) subunit gene of EHEC O157∶H7.Methods: A pair of primers were designed based on the Stx2B subunit gene sequence of EHEC O157∶H7.The Stx2B gene was amplified from the EHEC O157∶H7 chromosome by PCR and cloned into the pMD18-T vector.Thereafter,the gene was cut from the pMD18-T vector and cloned into the prokaryotic expression plasmid pET-28a vector.Then the recombinant plasmids were transformed into E. coli BL21(DE3) and the transformed host strain induced by IPTG.The expression protein was detected by SDS-PAGE and Western-blot analyses.Results: The Stx2B gene was successfully cloned into pMD18-T and pET-28a vectors,and the expressed protein identified by SDS-PAGE and Western blot.The molecular mass(Mr) of the expressed product was about 7 500,and the expression rate about 40%. Conclusion: The Stx2B gene was successfully cloned and effectively expressed in the prokaryotic expression system.