Volume 93  ·   Issue 44  ·  2013

To assess the efficacy and safety of 10-day sequential therapy for Helicobacter pylori (H.pylori) eradication in children.

The databases of Cochrane Central Register of Controlled Trials (CENTRAL,2013,N01), Medline, Embase, OVID,Chinese Biological Medicine database (CBMDisc), CNKI, Chinese VIP and WANFANG (all from 2000-2013) were searched.And manual searches were performed for the relevant journals and conference proceedings.Prospective, randomized controlled trials of 10-day sequential therapy for H.pylori eradication in children were selected.The systematic review was conducted with the method recommended by The Cochrane Collaboration.

Fifteen trials with a total of 1348 patients were included for analysis.The meta-analysis showed: (1)H.pylori eradication rates:10-day sequential therapy were superior to triple therapy (80.78% (521/645) vs 69.84% (491/703), OR 2.13, 95% CI: 1.62-2.80,P<0.01).It was also superior to 7-day and 10-day standard triple therapy (86.03% (271/315) vs 73.77% (225/305), 79.04% (181/229) vs 65.19% (176/270); OR 2.23, 2.11;95% CI:1.47-3.36,1.37-3.24;both P<0.01).There were non-superior to 14-day standard triple therapy (73.97% (108/146) vs 70.31% (90/128),OR 1.87, 95% CI:0.46-7.69, P=0.38). (2) Adverse effect rate:adverse events were similar between sequential and standard triple therapies (P=0.58).

The 10-day sequential therapy may be a new effective and safe option in treatment of H.pylori infections in children.

To evaluate the efficacy and tolerability of a furazolidone-based quadruple rescue regimen in patients with previously failed Helicobacter pylori (H.pylori) eradication treatment.

A total of 156 patients were recruited from July 2011 to June 2013 at Beijing Hospital and randomized into one of the following 10-day treatment regimens: (1) Esomeprazole 20 mg twice daily, furazolidone 100 mg twice daily, amoxicillin 1000 mg twice daily, bismuth salts 150 mg thrice daily for 10 days. (2) Esomeprazole 20 mg twice daily, levofloxacin 500 mg daily, amoxicillin 1000 mg twice daily, bismuth salts 150 mg thrice daily for 10 days.H.pylori status was re-assessed with the ¹³C-urea breath test at 4 weeks after the end of therapy.

The intentionto-treat (ITT) and per-protocol (PP) H.pylori eradication rates were 83.8% (67/80) and 88.2% (67/76) in the furazolidone-based quadruple group, and 69.7% (53/76) and 73.6% (53/72) in the levofloxacin-based quadruple group respectively (χ²=4.311, 5.100;P=0.038, 0.024).

The 10-day furazolidone-based quadruple therapy is an efficious rescue strategy in patients with previously failed eradication therapy.

To observe the efficacy of Jinghuaweikang gelatin pearls plus proton pump inhibitor (PPI)-based triple regimen in the treatment of chronic atrophic gastritis (CAG) patients with Helicobacter pylori (H.pylori) infection.

For this multicenter, randomized, controlled clinical study,90 patients of endoscopically confirmed CAG with positive H.pylori (¹³C or ¹⁴C-urea breath test (UBT) or rapid urease test) were enrolled.There were 46 males and 44 females with an age range of (54±10) years.None of them had H.py lori eradication background.They were randomly divided into 2 groups, Group LACJ (n=45) received lansoprazole 30 mg+ amoxicillin 1000 mg+ clarithromycin 500 mg+ jinghuaweikang gelatin pearls 240 mg, twice daily, for 10 days (d1-10) plus another 14 days (d11-24) only with jinghuaweikang gelatin pearls 240 mg, twice daily.Group LACB (n=45) had standard quadruple regimen treatment: lansoprazole 30 mg+ amoxicillin 1000 mg+ clarithromycin 500 mg+ bismuth potassium citrate 220 mg, twice daily for 10 days (d1-10).The status of H.pylori was detected by ¹³C-UBT at least 28 days after therapy.

The eradication rates in Groups LACJand LACB were as follows:per-protocol (PP) :70.5% (31/44) and 83.3% (35/42), intention-to-treat (ITT): 68.9% (31/45) and 77.8% (35/45)(both P>0.05).The symptomatic improvements of bloating in upper abdomen, belching and epigastric pain after treatment in both groups.And those in Group LACJ was higher than those of Group LACB, but no statistical difference existed between two groups (all P>0.05).

The efficacy of LACJ for the treatment of CAG patients with H.pylori infection is similar to LACB.And the symptomatic improvement of patients is better than LACB.

To evaluate the clinical efficacy of bio-mesh-reinforced pancreatogastrostomy.

A total of 23 patients undergoing bio-mesh-reinforced pancreatogastrostomy from May 2011 to January 2013 were retrospectively analyzed.Their demographic data, operative parameters and post-operative outcomes were recorded.The severity of pancreatic leak was determined according to the criteria of International Study Group on Pancreatic Fistula (ISGPF).

The mean anastomotic time was 24 (20-35) minutes.Intra-operative leak tests showed all pancreatic anastomoses were watertight.Six patients (26.1%) had pancreatic leakage of grade A.One patient (4.3%) had pancreatic leakage of grade B.No patient developed postoperative pancreatic leakage of class C.One case of abdominal infection was reported.No severe complications such as hemorrhage, bile leakage or gastrojejunostomy leakage were observed.All patients recovered well within Month 1 post-discharge.

This novel technique may be a simple and feasible strategy for all types of pancreatic remnants.

To explore the effective therapy of female overactive bladder unresponsive to behavior training.

A total of 67 patients with female overactive bladder unresponsive to behavior training were enrolled from January 2012 to January 2013 at Liaocheng Second People′s Hospital.They were randomized into trial and control groups (Ⅰand Ⅱ).Their mean age was 39.8 (19- 57) years.And the mean disease course was 3.8 (1-16) years.The trial group (n=24) received oral formulations of solifenacin succinate (5 mg, once a day) and naftopidil (25 mg, every evening).The control groupⅠ (n=22) had only solifenacin succinate and the control groupⅡ (n=21) only naftopidil.The treatment lasted for 4 weeks.The time of urination per day, average amount of mona-urination and maximum amount of mona-urination were observed.The changes of all parameters before and after treatment were assessed.And statistic analysis was performed.

The urinary urgency score of the trial, control groupⅠ and controlⅡ groups were 0.8±0.1, 1.8±0.8, 2.1±0.9; and the times of urination per day 9±4, 13±4, 14±5, average amount of monaurination (295±79), (211±67), (185±64) ml and maximum amount of mona-urination (352±88), (292±75), (235±69) ml respectively.These parameters showed significant differences between the trial and control groups (all P<0.05).

A combination of solifenacin succinate and naftopidil can effectively relieve the symptoms of female overactive bladder and improve the life quality.

To analyze the clinical characteristics and therapeutics of elderly patients with breast carcinomas.

A total of 312 patients (≥70 years old) admitted into Breast Center of Beijing Hospital from September 1997 to September 2011 were included for this retrospective study.Their clinical characteristics, treatment options and prognostic factors were analyzed.

They accounted for 14.6% (312/2137) of breast cancer patients treated during the same period.The predominant complaint was palpable mass (n=250, 80.1%).90.4% (n=282) of them were complicated with other diseases.The major pathologic type was invasive ductal carcinoma of non-special type (n=248, 79.5%).The positive rate of estrogen receptor (ER) and (or) progesterone receptor (PR) was 75.3% (n=235), and 15.7% (n=49) cases were Cerb-B₂(+++).The 5, 10-year overall survival were 75.0% and 41.8% respectively.TNM stage, vascular thrombosis, operative regimen and adjuvant therapy were prognostic factors.

Surgical operation and endocrine therapy are the first choice remedies for elderly patients.And the major prognostic factors are TNM stage, vascular thrombosis and treatment regimens.

To evaluate the value of urine Livin and Survivin m RNA expression in the early diagnosis of bladder cancer.

Fifty-two cases of early bladder cancer and 30 cases of non-urinary system tumors were selected for the combined detection of urinary Livin and Survivin and urine cytology.

Livin and Survivin in urine and urinary cytology sensitivity were 71.2% (37/52), 67.3% (35/52), 23.1% (12/52);Specificity were 96.7% (29/30), 93.3% (28/30) and 96.7% (29/30) respectively.Urine Survivin sensitivity, specificity compared with Livin were no significant difference (all P>0.05).Urinary Livin and Survivin were more sensitive than urine cytology,and the differences were statistically significant (all P<0.05).

The combined urinary detection of Livin, Survivin mRNA expression is a noninvasive early diagnosis of bladder cancer with sensitivity and specificity.

To evaluate the values of combined detection method of EB viral Rta-IgG, VCA-IgA, EA-IgA and EB viral DNA in the diagnosis of nasopharyngeal carcinoma (NPC).

Serum and plasma samples from 131 untreated NPC patients, 52 non-NPC patients with NPC-like symptoms and 148 healthy donors from January to December 2012 were collected.Immunoenzymatic staining was used to detect VCA-IgA and EA-IgA in sera.ELISA was performed to detect Rta-IgG antibody in sera and real-time fluorescent quantitative PCR for measuring EBV DNA in plasma.The clinical characteristics of 3 groups were compared.ROC curve and correlation analyses were performed to assess the detection assays for the diagnosis of NPC.

The positive rates of EBV Rta-IgG, VCA-Ig A, EA-Ig A and EBV DNA in untreated NPC patient group were higher than those in other two groups.The differences were statistically significant (all P<0.01).The differences of Rta-Ig G antibody positive rates, EBV-DNA levels and EBV-DNA positive rates at different clinical stages were statistically significant (all P<0.05).The positive rates of VCA-IgA and EA-IgA were not related with clinical stages (P>0.05).Areas under ROC curve for Rta-IgG and EBV-DNA were 0.901 and 0.827 respectively.All four diagnostic assays demonstrated excellent efficiency.The sensitivity and specificity of individual assays were as follows: Rta-IgG: 77.9%,92.5%; VCA-IgA 93.1%, 91.5%; EA-IgA: 74.8%, 99.5%; EBV-DNA 64.9%, 97.0%.The sensitivity and specificity of combined assays were 97.7% and 83.5% respectively.

Combined detection method of EB viral Rta-IgG, VCA-IgA, EA-IgA and EB viral DNA are efficient for the diagnosis of NPC.

To evaluate the relationship and strength of association for alcohol drinking and tea consumption with the riskS of nasopharynx cancer among Chinese population so as to provide control rationales for nasopharynx cancer in China.

A systematic search of 3 Chinese electronic databases (CNKI, VIP, Wanfang) and 3 English databases (Pubmed, ScienceDirect and Springer Link) up to March 2013 was performed.Two reviewers independently conducted the literature search, examined eligibility and performed data extraction and quality evaluations.Pooled odd ratio (OR) value and 95% CI value were calculated with random-effects model weighted with inverse of variances.

A total of 14 studies (including 3 cohort and 11 casecontrol) involving 6559 cases of nasopharynx cancer and 10 567 controls from 6 provinces were included.The pooled OR between alcohol drinking and risks of nasopharynx cancer was 1.12 (95% CI: 0.98-1.26; I²=44.5%, P=0.037).Compared with the non-drinkers, the risks of nasopharynx cancer for regular drinkers and occasional drinkers were 1.18 (95% CI:1.00-1.38; I²=0.0%,P=0.578) and 0.76 (95% CI:0.65-0.89; I²=33.4%,P=0.212).And the association of tea consumption with the risks of nasopharynx cancer was 0.53 (95% CI: 0.43-0.60; I²=17.9%, P=0.301).

In China, occasional alcohol drinking may decrease the risks of nasopharynx cancer while regular drinking elevates the risks.And there is significantly protective effect for tea consumption on the risks of nasopharynx cancer.

To evaluate the significance of magnetic resonance imaging (MRI) T²value analysis in patients with iron overload and compare it with other clinical parameters.

A total of 53 patients with suspected iron overload were recruited from four Beijing hospitals from December 2010 to December 2012.Their liver and heart T²values were calculated and their serum ferritin (SF), transferin saturation, blood transfusion volume and other clinical parameters were recorded and analyzed.Results There were 37 males and 16 females with a medium age of 50 years (15-72 years).Their etiologies included myelodysplastic syndromes (MDS, n=25), aplastic anemia (AA,n=16), myelofibrosis (n=5), hemachromatosis (n=2) andβthalassaemia (n=2),and 3 patients with high SF values were found on regular health examinations.Among them, there were transfusion history (n=45), SF>1000μg/L (n=49), sign of iron overload (n=10), abnormal liver function (n=38) and hyperglycemia (n=32).T² value analysis showed that 10 patients had no evidence of iron overload, 43 patients had liver iron overload (14 mild, 22 moderate and 7 severe) and 2 patients had heart iron overload (1 MDS with heavy transfusion history and 1 AA with heart failure).No relations existed between T²value and SF (P=0.050), T² value and transfusion volume (P=0.820), and liver T² value and heart T² value (P=0.129).

MRI T² value is an accurate way of quantitative detection of iron overload.It provides a comprehensive understanding of patients with iron overload in conjunctions with MRI T²value and other clinical parameters.

To investigate the role of autophagy in the injury of Hep G-2 cells induced by hepatitis B virus x protein (HBx).

After HBx transfection, the cells were used to detect the formation of autophagosomes and observed by transmission electron microscopy, monodansylcadaverine (MDC) staining autophagic vacuole (AV), immunofluorescen ce staining microtubule-associated protein light chain 3 (MAP1-LC3) protein, and Western blotting examining the ratio of LC3-Ⅱ/LC3-Ⅰ (gray level: 0.760±0.078 vs 0.520±0.086,P<0.05), beclin 1 (gray level:0.875±0.093 vs 0.220±0.087,P<0.05) and lysosome associated membrane protein 2a (lamp2a) protein (gray level:0.320±0.061 vs 0.120±0.064,P<0.05) levels.

(1) HBx transfected upregulated the expression of LC3-Ⅱ, LC3-Ⅰ, beclin 1 and lamp2a protein. (2) HBx transfected brought about an increase in the formation of autophagosomes and autolysosomes.

HBx activates the autophagic lysosome pathway in HepG-2 cells through the LC3/beclin1 pathway.

To explore the regulation of autophagy-lysosome pathway (ALP) in skeletal muscle of diabetic nephropathy and examine the effect of low protein diet plusα-keto acid on ALP.

A total of 45 24-week-old Goto-Kakizaki rats were randomized to receive normal protein (22%) diet (NPD), low-protein (6%) diet (LPD) or low-protein (5%) plusα-keto acids (1%) diet (Keto)(n=15 each).Wistar control rats had a normal protein diet.The mRNA and protein levels of ALP markers LC3B, Bnip3, Cathepsin L in soleus muscle were evaluated at 48 weeks.Electron microscopy was used to confirm the changes of autophagy.

Compared with CTL group, the mRNA levels of LC3B, Bnip3, Cathepsin L in soleus muscle of rats on NPD were higher, and protein levels of LC3B-Ⅰ,LC3B-Ⅱ, Bnip3, Cathepsin L in soleus muscle of rats on NPD also higher than CTL group (0.82±0.33 vs 0.25±0.07,0.76±0.38 vs 0.20±0.12,1.25±0.30 vs 0.56±0.19, 1.29±0.40 vs 0.69±0.20).The mRNA levels of LC3B, Bnip3 and Cathepsin L in LPD group were slightly lower, compared with NPD group.However there was no statistical significance.Similarly the protein levels of LC3B-Ⅰ, LC3B-Ⅱ, Bnip3 and Cathepsin L in LPD group were slightly lower with no statistical significance.In contrast, the mRNA levels of LC3B, Bnip3 and Cathepsin L were greatly lower in Keto group in comparison with NPD and LPD.And protein levels of LC3B-Ⅰ,LC3B-Ⅱ, Bnip3 and Cathepsin L were also greatly lower in Keto group in comparison with NPD and LPD.Additionally, autophagosome or autolysosome was found in NPD and LPD groups by electron microscopy.

ALP is activated in skeletal muscle of diabetic nephropathy rats.And low protein plusα-keto acid decrease the activation of ALP and improve muscle wasting.

To explore the relation between high-fat diet and pulmonary fibrosis and the inhibition of N-acetylcysteine (NAC) on lung tissue in rats.

Thirty Sprague Dawley (SD) male rats were randomly divided into control group (n=10) on a quantitative control Lieber-DeCarli liquid diet,a high-fat diet group (n=10) on a high fat-diet Lieber-DeCarli liquid diet and NAC group (n=10) on NAC 300 mg·kg^-1·d^-1.All rats were sacrificed 8 weeks later.The morphological changes and collagen deposition in lung tissue were observed by hematoxylin& eosin and Masson staining while the contents of glutathione (GSH) and haluronic acid (HA) measured through colorimetry and enzyme-linked immunosorbent assay.And the expression of transforming growth factor-β₁ (TGF-β₁) expression in lung tissue was detected through immunohistochemistry.

There were variable degree of alveolar and alveolar septal infiltration of inflammatory cells.And more deposition of collagen fibers appeared at intervals of alveolar in high fat group.Similar pathological changes were found in NAC group, but the degree was lower than that of high-fat group.Compared to the control and NAC groups, the lung tissue content of GSH decreased (GSH: 0.11±0.05 vs 0.19±0.11, 0.22±0.14 mg/g, both P<0.05) while HA and TGF-β₁increased in high-fat diet group (HA:0.57±0.06 vs 0.46±0.04,0.41±0.04 mg/g; TGF-β₁:24.6±3.4 vs 16.8±2.6, 11.7±1.5, all P<0.05).

High-fat diet may induce pulmonary fibrosis in rats and NAC has inhibitory effects.

To observe the effect of PKCδon the phosphorylation of p66Shc and its mitochondrial translocation in human proximal tubular cells (HK-2) under high glucose (HG).

HK-2 cells were incubated with different concentrations of D-glucose (5-45 mmol/L) for indicated time (0-48 h).Then the mRNA expressions of PKCδand p66Shc and the phosphorylation levels of PKCδ (p-PKCδ) and p66Shc (p-p66Shc) were determined by real-time polymerase chain reaction (PCR) and Western blot analysis respectively.In addition, the effect of PKCδinhibitor on the phosphorylation and mitochondrial translocation of p66Shc in HK-2 cells exposed to HG was also observed.HK-2 cells were divided into 3 groups of 5 mmol/L glucose, 30 mmol/L glucose and 30 mmol/L glucose+1.0μmol/L Rottlerin.Cell immunofluorescence and Western blotting were used to observe the phosphorylation and mitochondrial translocation of p66Shc.

Both mRNA expression and phosphorylation level of p66shc and PKCδsignificantly increased in HK-2 cells after exposure to HG (15,30,45 mmol/L).And it was in a concentration- and time-dependent manner as compared with control group (up-regulated 0.9, 1.3 and 1.6-fold in mRNA of PKCδ, 0.4,1.5 and 2.0-fold in protein of p-PKCδrespectively (all P<0.05).PKCδinhibitor Rottlerin dramatically inhibited the phosphorylation and mitochondrial translocation of p66Shc induced by HG in HK-2 cells (down-regulated 3.1 folds in protein of p-p66Shc in mitochondria, P<0.01).

HG increases the transcription and phosphorylation of PKCδand p66Shc in HK-2 cells.And PKCδmay modulate the phosphorylation and mitochondrial translocation of p66Shc under HG.

To perform a pharmacogenetic study of a patient with warfarin hypersensitivity and determine the biological function of a novel CYP2C9 mutation.

Genomic DNAs were extracted from the warfarin hypersensitive patient and used for genetic screening of CYP2C9, VKORC1 and CYP4F2 by direct sequencing.Acquired sequences were aligned and blasted with public nucleotide database to clarify whether site mutations were present in these 3 genes.Full-length cDNA fragments of CYP2C92,3,13 and 1400T>C were obtained by polymerase chain reaction (PCR) site-directed mutagenesis and used for the insect expression vector construction.According to the manufacturer′s instruction, insect cell microsomes expressing 5 CYP2C9 variants were obtained with Bac-to-Bac Baculovirus Expression System.Then CYP2C9 protein expression level for each variant was quantified by Western blot and tolbutamide used as a probing substrate for assessing the in vitro metabolic characteristics of each CYP2C9 variant. V_max and K_m values were calculated and used for evaluating the biological impact of novel 1400T>C mutant.

One novel nucleotide mutation 1400T>C was detected in DNAs of tested patient.And this site mutation resulted in one amino acid change at position 467 of CYP2C9 protein (L467P).Other two candidate genes VKORC1 and CYP4F2 were found to be the wild-type alleles.When expressed in insect cell microsomes, the relative clearance values of CYP2C92,3,13 and L467P variants to tolbutamide were 91.58%, 13.55%, 0.11% and 1.59% to that of wild-type protein respectively.

1400T>C mutation of CYP2C9 gene greatly reduces the in vitro enzymatic activity of CYP2C9.While taking the drugs metabolized by CYP2C9, the patients carrying this mutated allele should take more cautions because their metabolic rates are slower than those of wild-type carriers.Ensuing drug accumulation in vivo may cause potentially serious adverse reactions.

To explore whether the signal pathways of phosphoinositide 3-kinase (PI3K) and Notch can realize coordinated regulation on the activation and proliferation of CD4⁺T lymphocytes.

Male BALB/c mice were randomly divided into control and asthma groups.Then the murine model of asthma was established by the method of ovalbumin (OVA) challenge.The CD4⁺T lymphocytes were isolated by magnetic activated cell sorter (MACS) and then activated with phytohaemagglutinin (PHA)(10μg/ml) and IL-2 (1000 U/ml) for 6 h.Those cells were then divided into Group A: without any treatment; Group B:treatment with PI3 K inhibitor (LY294002); Group C:treatment with Notch inhibitor (gamma-secretase inhibitor, DAPT); Group D: treatment with PI3K inhibitor and Notch inhibitor.The protein and transcription levels of Cyclin A, Cyclin D1 and P27^kip1of CD4⁺T lymphocytes were assessed by flow cytometry and reverse transcriptase polymerase chain reaction (RT-PCR).

The results of flow cytometry showed that the purity of MACS-isolated CD4⁺T lymphocytes was 90.0%±5.2% and the survival rate 94.8%±3.2%.The protein (28.0%±3.5%,14.9%±3.4%) and mRNA levels (0.55±0.16,1.38±0.42) of Cyclin A and Cyclin D1 in CD4⁺T lymphocytes of asthma group were significantly higher than those of the control group (13.4%±3.5%,7.7%±1.8% and 0.32±0.10,0.92±0.37) (P=0.002,0.036 and P=0.007, 0.042).The protein and mRNA levels (23.3%±3.9% and 0.16±0.03) of P27^kip1of asthma group were significantly lower than those of control group (37.5%±5.8% and 0.32±0.03, P=0.006 and P=0.000).The protein and mRNA levels of Cyclin D1 in groups A, B, C and D-treated CD4⁺T lymphocytes were 12.2%±3.7%, 7.3%±3.0%, 8.1%±2.3%, 4.2%±1.7% and 1.71±0.44, 1.07±0.31, 1.21±0.32 and 0.62±0.20 respectively; groups B, C and D decreased markedly compared with group A (all P<0.01) while group D decreased significantly compared with groups B and C (all P<0.05).The protein levels of P27^kip1in groups A, B, C and D were 22.9%±3.0%, 31.6%±5.3%,28.4%±5.6% and 44.6%±2.8% respectively; group B was significantly higher than that of group A (P=0.016) while group D was significantly higher than those of groups A, B and C (P=0.003, 0.004, 0.000).Meanwhile P27^kip1 mRNA levels in each group were 0.16±0.07, 0.36±0.09, 0.63±0.08 and 0.99±0.21 respectively; groups B, C and D were much higher than that of group A (P=0.016, 0.000, 0.000) while group D was significantly higher than those of groups B and C (P=0.000, 0.023).The protein and m RNA levels of Cylin A showed no statistical significance among different experimental groups (all P>0.05).

The signal pathways of PI3K and Notch may coordinately up-regulate the expression of positive regulatory factor cylin D1 and down-regulation the expression of negative regulatory factor P27^kip1of CD4⁺T lymphocytes.