Objective: To investigate the effects of Liuwei Dihuang decoction (LW), its San-bu (three tonics) and Sanxie (three eliminators) ingredients on regulating the mmune function of senescence-accelerated mice (SAM). Methods: SAM-prone (SAMP) 8 mice were orally administrated with LW (10 g/kg), San-bu (6.4 g/kg) and San-xie (3.6 g/kg) respectively, once per day for 60 days, while SAM-resistant ( SAMR) 1 mice administrated with distilled water as control group. 3H-TdR incorporation was applied for detecting splenocyte proliferation. Flow cytometry (FCM) was used to observe the percentage of CD3+, CD4+,CD8+ and CD19+ ymphocytes in the spleens. Results: Compared with SAMR1 group, the splenocyte proliferation induced by ConA an LPS, percentages of CD3+, CD4 +, CD8+ and CD19+ lymphocytes and ratios of CD3+/CD19+ and CD4+/CD8+ in spleens decreased, while percentage of CD19+ lymphocytes increased in SAMP8 group. After administration of LW, San-bu and San-xie, the above ndexes were mproved to various degrees. LW showed better effects on improving LPS-induced splenocyte proliferation, percentages of CD3+, CD19+ and CD4+ lymphocytes, and the ratios of CD3+ /CD19+ and CD4+/CD8+ than San-bu and San-xie. San-bu showed better effect on mproving ConA-induced splenocyte proliferation than LW and San-xie, and better effect on mproving the percentages of CD3+, CD19+ and CD4+ lymphocytes in the spleens and the ratios of CD3+/CD19+ than San-xie, while San-xie had better effect on regulating CD4+/CD8+ ratio than San-bu. Conclusion: These results suggest that LW regulates the function of T and B lymphocytes and the mbalance of subsets of CD4+ and CD8+ lymphocytes in the spleens of SAMP8. LW shows better effects than San-bu or San-xie used alone, while San-bu and San-xie have their particular effects. The effects of LW on regulating the immune function might be the integral results of San-bu and San-xie. The results provid some experimental evidences for revealing the compatibility of LW.

To establish single- and double-transfected transgenic cells stably expressing hMATE1, hMATE1 cDNA was cloned by RT-PCR from human cryopreserved kidney tissue, and subcloned into pcDNA3.1(+) plasmid by virtue of both HindIII and Kpn I restriction enzyme sites. Subsequently, the recombined pcDNA3.1(+)- hMATE1 plasmid was transfected into MDCK, MDCK-hOCT1 or MDCK-hOCT2 cells using Lipofectamine 2000 Reagent. After a 14-day-cultivation with hygromycin B at the concentration of 400 µg · mL(-1), all clones were screened with DAPI and MPP+ as substrates to identify the best candidate. The mRNA content of hMATE1, the cellular accumulation of metformin with or without cimetidine as inhibitor, or transportation of cimetidine was further valuated. The results showed that all of the three cell models over expressed hMATE1 mRNA. The cellular accumulation of metformin in MDCK-hMATE1 was 17.6 folds of the control cell, which was significantly inhibited by 100 µmol · L(-1) cimetidine. The transcellular transport parameter net efflux ratios of cimetidine across MDCK-hOCT1/hMATE1 and MDCK-hOCT2/hMATE1 monolayer were 17.5 and 3.65, respectively. In conclusion, cell models with good hMATE1 function have been established successfully, which can be applied to study the drug transport or drug-drug interaction involving hMATE1 alone or together with hOCT1/2 in vitro.
Animals ; Biological Transport ; Cimetidine ; pharmacology ; DNA, Complementary ; Dogs ; Drug Interactions ; Humans ; Madin Darby Canine Kidney Cells ; Metformin ; pharmacology ; Organic Cation Transport Proteins ; genetics ; metabolism ; Transfection

Objective: To investigate the sonographic features of appendicitis in preterm infants. Methods: A total of 28 cases of premature infants with acute appendicitis diagnosed by bedside abdominal ultrasound in the First Hospital of Jilin University from November 2012 to January 2019 were recruited. Basic clinical information, abdominal ultrasound images, surgical results, management and outcomes were collected and analyzed. Descriptive statistical methods were used for data analysis. Results: Among the 28 cases, 21 (75.0%) were males and seven (25.0%) were females. All of them were diagnosed as having acute appendicitis with perforation according to the bedside ultrasound. Five (17.8%) presented direct signs of appendicitis, i.e. partial structure of the appendix and perforation site. The other 23 (82.2%) showed indirect signs, including heterogeneous echotexture or hypoechoic patterns between the liver and right kidney in six cases, heterogeneously hypoechoic areas between the bowels in the right lower abdomen in seven cases, and dissociative effusion between the bowels in the right lower abdomen with poor sound transmission and disorder of surrounding intestinal structure in ten cases. Twenty-one out of the 28 cases (75.0%) exhibited bowel wall thickening at right lower abdomen, absence of intestinal peristalsis and effusion echoes between the intestines with poor sound transmission. Emergent surgeries were performed and diagnoses of appendicitis with perforation were confirmed. All the 21 cases were discharged after full recovery. Seven cases (25.0%) showed confined cystic images and received conservative treatment. One of them developed adhesive intestinal obstruction during follow-ups and underwent surgical treatment, during which local formations of wrapping after appendiceal perforation and obstruction due to surrounding intestinal adhesion were observed. The other six cases recovered after conservative management with gradually reduced peritoneal effusion, normal omental echo patterns and improved inflammatory indicators and abdominal symptoms, and no ileus occurred during follow-ups after discharge. Conclusions Symptoms of appendicitis in preterm infants are non-specific, and perforation is more likely to be seen. Bedside ultrasonography mainly shows indirect signs of appendicitis, and direct signs in some infants. Bedside ultrasound can be an essential tool for the diagnosis of these conditions with high accuracy.

OBJECTIVETo construct the KbvR gene of LuxR family deletion mutant and complementation strains from Klebsiella pneumoniae NTUH-K2044 and analyze the effect of KbvR on bacterial growth, biofilm formation and capsular synthesis.

METHODSA KbvR gene deletion mutant strain was constructed using the suicide vector pKO3-Km, and the gene fragment including KbvR coding region, promoter area and transcription termination area were amplified and cloned into pGEM-T-easy plasmid to construct KbvR complementation strain. The growth curves of the wild-type strain, KbvR gene deletion mutant strain and complementation strain were observed to assess the effect of KbvR on bacterial growth. Crystal violet staining method was used to measure the effect of KbvR on biofilm formation; the effect of KbvR on capsular synthesis was detected using string test, centrifugal test and RT-PCR.

RESULTSThe KbvR deletion mutant and complementation strains were constructed successfully. KbvR gene did not affect the growth of the bacteria, but biofilm formation and capsular synthesis were attenuated in KbvR deletion mutant strain.

CONCLUSIONAs a transcription factor of the LuxR family orphans of the quorum sensing system, KbvR positively regulates bacterial biofilm formation by affecting capsular synthesis.

Bacterial Capsules ; Bacterial Proteins ; metabolism ; Biofilms ; Gene Deletion ; Klebsiella pneumoniae ; cytology ; Plasmids ; Promoter Regions, Genetic

Aim To investigate the molecular mecha-nism of mTORC1/2 inhibitor PP242, which inhibiting cholangiocyte cell preliferation and cystic diliatation via inducing apoptosis and autophagy in the polycystic kid-ney ( PCK ) rats. Methods The expression of p-mTOR and p-Akt in the bile duct epithelial cells was examined by immunohistochemistry. The inhibiting effect of rapamycin and PP242 on cell proliferation ac-tivity on bile duct epithelial cells, the effect of gene si-lence on LC3, Beclin-1 and the effect of the authoph-agy-specific inhibitor 3-methyladenine (3-MA) on cell proliferation were respectively analyzed by WST-1 as-say. The expression of PI3K/Akt signaling pathway re-lated proteins, autophagy-related proteins LC3, Bec-lin-1 and clevead caspase-3, which were treated by PP242 were determined by Western blot. The effect of PP242 on apoptosis was detected by Annexin V/PI double staining and ELISA. The expression of LC3 in cytoplasm was detected by immunofluorescence. The a-bility of rat bile duct epithelial cells spheroid formation was detected by 3D cell culture method, and the cells were treated by single applied with rapamycin and ap- plied rapamycin combined with Rictor gene silencing respectively. Results The protein levels of p-Akt and p-mTOR markedly increased in the bile duct epitheli-um of PCK rats. PP242 inhibited the proliferation of bile duct epithelial cells more effectively than rapamy-cin and showed a dose-and time-dependent manner ( P<0.05 ) . PP242 significantly reduced the levels of PI3K/Akt signaling pathway-related proteins in PCK rat cholangiocytes. PP242 induced apoptosis and auto-phagy, up-regulated the levels of cleaved caspase-3, Beclin-1 and increased the ratio of LC3-II/LC3-I. The combination of Rictor gene silencing and rapamycin was more effective than rapamycin alone in inhibiting cholangiocytes in PCK rats. The inhibitory effect of PP242 on the cell viability was significantly weakened by treatment with 3-MA and knockdown of LC3 and Beclin-1 ( P <0.05 ) . Conclusions PP242 inhibits the proliferation of PCK rat cholangiocytes through PI3K/Akt/mTOR signaling pathway, and the mecha-nism is closely related with autophagy and apoptosis.

OBJECTIVETo describe the distribution changes of the mortality rate for cervical cancer in China between the 1970's and 1990's and provide the scientific evidence for the prevention and control strategies for cervical cancer campaign in China between next century.

METHODSData from two National Surveys for the Causes of Death in 1970's and 1990's in China. The crude and adjusted mortality rates for the cervical cancer and the distributions based on age and area were calculated and described. The comparison of the differences of changes between two mortality rates periods and together with its trends were shown based on the age-standardized.

RESULTSDuring two decades, the mortality rate for cervical cancer was 10.7 per 100,000 in 1970's which declined to 3.89 per 100,000 in 1990's, and from the 3rd ranking among all female malignant tumors to the 6th in 1990's (decreased about 63.64%). But the declination was not evenly. There have still been some high-risk areas, most located in rural countries in the mid-west of China, with rates remain unchanged and even at the highest level in the world, such as Wudu in Gansu and Yangcheng in Shanxi. A big difference was showed between rural country and city, but in both of them, the mortality rates in 1990's were significantly much lower than in 1970's (P = 0.001) at each five-year age group. And in the city, there was a much sharper increased trend in young women in 1990's.

CONCLUSIONSThe mortality rate for cervical cancer campaign in China has been substantially declined during past twenty years, but it's still a major health problem for women, especially in rural China. The focus of the prevention and control for the cervical cancer in the next century should put on rural areas, especially in mid-west of China and young women in the city.

Adult ; Age Factors ; Aged ; Cause of Death ; trends ; China ; epidemiology ; Female ; Humans ; Middle Aged ; Retrospective Studies ; Rural Health ; Uterine Cervical Neoplasms ; mortality ; prevention & control

AIMTo study the chemical composition of Tripterygirm wilfordii Hook. f.

METHODSColunm chromatography was used to separate the chemical constituents. UV, IR, MS, HRMS, 1HNMR, 13CNMR (COM and OFR), 1H-1H COSY, 1H-13C COSY, NOESY and COLOC spectra were used to determine the structures of the isolated constituents.

RESULTSTwo sesquiterpene alkaloids were isolated and their structures were elucidated as wilforgine and wilfordlongine on the basis of spectral evidence.

CONCLUSIONWilfordclonine is a new sesquiterpene alkaloid.

Lactones ; chemistry ; isolation & purification ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Pyridines ; chemistry ; isolation & purification ; Tripterygium ; chemistry

OBJECTIVETo investigate the expression of translationally controlled tumor protein (TCTP) in human breast cancer tissues and its clinical significances.

METHODSThe expression of TCTP in 94 human breast cancer and the corresponding adjacent normal mammary tissues were detected using immunohistochemistry.

RESULTSThe expression rate of TCTP was 64.89% in human breast cancer tissues, significantly higher than that in normal benign mammary tissues (39.36%, P<0.001). TCTP overexpression was positively correlated to the tumor size, clinical stage, lymph node metastasis and histological grade of breast cancer (P<0.05). Patients with positive TCTP expression had a significantly shorter mean survival time than those with negative expression (P<0.001).

CONCLUSIONTCTP may play an important role in the tumorigenesis and development of breast cancer, and can be an important prognostic factor for this malignancy.

Adult ; Aged ; Biomarkers, Tumor ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; Female ; Humans ; Middle Aged

Objective To investigate the pathogenic mutations of BRCA1 and BRCA2 in patients with early-onset breast cancer(≤35 years) and explore the relationships between BRCA1/2 mutations and clinical features.Methods Seventy-four patients with early-onset breast cancer were enrolled,who were treated in Hospital 307 between September 2014 and June 2016.High-throughput sequencing was used to test the 49 exon sequences and adjacent sequences of BRCA1 and BRCA2.χ2 test was used to analyze the distribution of BRCA1/2 pathogenic mutations in each group that was set up according to clinical features.Results Fifteen mutations(20.27%) were identified,including 5(6.76%) in BRCA1 and 10(13.51%) in BRCA2.Eleven new pathogenic mutations were discovered,and BRCA1:c.5470_5477delTGCCCAAT was found in one patient.The frequency of BRCA1/2 mutations in the group with a family history of breast cancer or ovarian cancer was higher than in the group without a family history (40.91% vs 11.54%) (χ2=6.534,P=0.011).Conclusion BRCA1/2 pathogenic mutation is significant for early-onset breast cancer,especially for those with a family history of breast or ovarian cancer.The new mutations may be specific to Chinese people.BRCA1:c.5470_5477delTGCCCAAT may be the ancestor mutation among the Chinese.

Objective To analyze the differences in the expression levels of the lncRNA MALAT1, NEAT, NEAT2 in peripheral blood mononuclear cell (PBMC) from tuberculosis patients and healthy controls. Methods We detected the lncRNA expression levels in PBMC from 79 tuberculosis patients and 82 healthy controls by quantitative reverse transcription polymerase chain reaction, and analyzed the correlation between lncRNA expression levels and some clinical features and laboratory indicators in tuberculosis patients. Results The expression levels of MALAT1, NEAT1 in PBMC of tuberculosis patients were significantly higher than healthy controls (Z=-4.386, P<0.001; Z=-10.175, P<0.001). There was no significant difference in the expression of NEAT2 between tuberculosis patients and healthy controls (Z=-0.203，P=0.839). The correlation results of lncRNA levels and some clinical features, laboratory indicators in tuberculosis patients suggested that the NEAT2 level in PBMC of newly treated tuberculosis patients was higher than recurrent tuberculosis patients, while the NEAT2 level in PBMC of sputum smear positive tuberculosis patients was lower than that of sputum smear negative tuberculosis patients (all P<0.05). There was a negative correlation between MALAT1 level and erythrocyte sedimentation rate (rs=-0.256, P=0.034). Conclusion MALAT1 and NEAT1 are abnormally expressed in PBMC of tuberculosis patients, and may be involved in the pathogenesis of pulmonary tuberculosis.