Breeding for low cadmium accumulation cereals / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Cadmium (Cd) is an element that is nonessential and extremely toxic to both plants and human beings. Soil contaminated with Cd has adverse impacts on crop yields and threatens human health via the food chain. Cultivation of low-Cd cultivars has been of particular interest and is one of the most cost-effective and promising approaches to minimize human dietary intake of Cd. Low-Cd crop cultivars should meet particular criteria, including acceptable yield and quality, and their edible parts should have Cd concentrations below maximum permissible concentrations for safe consumption, even when grown in Cd-contaminated soil. Several low-Cd cereal cultivars and genotypes have been developed worldwide through cultivar screening and conventional breeding. Molecular markers are powerful in facilitating the selection of low-Cd cereal cultivars. Modern molecular breeding technologies may have great potential in breeding programs for the development of low-Cd cultivars, especially when coupled with conventional breeding. In this review, we provide a synthesis of low-Cd cereal breeding.

Effect of Jiawei Changfengyin with Retention Enema Therapy on Th1/Th2 in Patients with Acute Radiation Proctitis / 中国实验方剂学杂志

Objective: To discuss the efficacy of Jiawei Changfengyin with retention enema therapy for acute radiation proctitis and investigate its influence on regulating levels of helper T cell(Th)1/Th2. Method: One hundred and twenty-eight patients were randomly divided into control group and observation group by random number table. Patients in control group (64 cases) got montmorillonite powder (3.0 g), dexamethasone (10 mg) and normal saline (100 mL), with retention enema therapy. Patients in observation group (64 cases) got Jiawei Changfengyin(Changfengyin+Xileisan+pearl powder), with retention enema therapy. Both groups of patients received enema once every night, and the treatment course was 4 weeks in both groups. Before and after treatment, scores of main symptoms, proctoscopy, routine examination of stool+occult blood, and KPS scores of quality of life were graded. Levels of Th1 cell factors[interleukin(IL)-1β, IL-8 and interferon-γ(IFN-γ)], Th2 cell factors (IL-4 and IL-10) were detected. Result: The total effective rate for comprehensive clinical efficacy was 95.31% in observation group, higher than 82.81% in control group (χ2=5.132, PPZ=2.764, PPPβ, IL-8 and IFN-γ in observation group were lower than those in control group, while levels of IL-4 and IL-10 were higher than those in control group (PConclusion: Jiawei Changfengyin can relieve symptoms of acute radiation proctitis, promote healing of rectal mucosa, improve quality of life, and regulate Th1/Th2 cytokines, with good repairing effect for intestinal mucosa.

Clinical features and prognosis of gastrointestinal injury due to foreign bodies in the upper gastrointestinal tract in children: a retrospective analysis of 217 cases / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the clinical features and prognosis of gastrointestinal injury caused by foreign bodies in the upper gastrointestinal tract in children.</p><p><b>METHODS</b>A retrospective analysis was performed for the clinical data of 217 children who were diagnosed with foreign bodies in the upper gastrointestinal tract complicated by gastrointestinal injury by gastroscopy from January 2011 to December 2016, including clinical features, gastroscopic findings, complications, and prognosis.</p><p><b>RESULTS</b>Among the 217 children, 114 (52.5%) were aged 1-3 years. The most common foreign body was coin (99/217, 45.6%), followed by hard/sharp-edged food (45/217, 20.7%) and metal (35/217, 16.1%). The most common gastrointestinal mucosal injury was ulceration (43.8%), followed by erosion (33.2%). Compared with other foreign bodies, button cells were significantly more likely to cause esophageal perforation (P<0.01). The esophagus was the most commonly injured organ (207/217, 95.4%). Of all the 217 children, 24 (11.1%) experienced infection. The children with perforation caused by foreign bodies had a significantly higher incidence rate of infection than those with ulceration caused by foreign bodies (P=0.003). Of all the 217 children, 204 (94.0%) underwent successful endoscopic removal of foreign bodies. Among these children, 98 were hospitalized due to severe mucosal injury and were given anti-infective therapy, antacids, and supportive care including enteral nutrition through a nasogastric tube and/or parenteral nutrition. Of all the children, 10 left the hospital and were lost to follow-up, and all the other children were improved and discharged.</p><p><b>CONCLUSIONS</b>Most cases of foreign bodies in the upper gastrointestinal tract occur at 1-3 years of age. Coin, hard/sharp-edged food, and metal are the most common foreign bodies. Button cells are more likely to cause esophageal perforation. The incidence rate of secondary infection increases with the increasing severity of gastrointestinal mucosal injury. Children undergoing endoscopic removal of foreign bodies and enteral nutrition through a nasogastric tube tend to have a good prognosis.</p>

Pancreatitis as the initial manifestation and abdominal lymph node enlargement in a boy / 中国当代儿科杂志

A boy aged 14 years had abdominal pain as the major manifestation, with elevated serum amylase and lipase. Abdominal ultrasound performed early after onset in another hospital showed enlargement of the pancreas and a reduction in echo. Magnetic resonance cholangiopancreatography (MRCP) showed pancreatic duct dilation and an unclear image of the head of the pancreas. Acute pancreatitis was considered. However, his symptoms were not relieved after fasting, fluid infusion, anti-acid therapy, and somatostatin therapy. Then, abdominal CT scan and MRCP found multiple low-density lesions of the pancreas and enlargement of the hilar and retroperitoneal lymph nodes. Exploratory laparotomy found pancreatic edema and multiple hilar nodules with unclear boundaries, and pathological biopsy showed anaplastic large-cell lymphoma. Since the liver, the spleen, bone marrow, and the central nervous system were not involved, he was diagnosed with stage III primary pancreatic lymphoma. After vindesine and dexamethasone were used to reduce tumor load, the patient underwent vindesine-pirarubicin-asparaginase-dexamethasone chemotherapy once and vinorelbine-dexamethasone chemotherapy 8 times. Imaging examination still showed multiple low-density lesions of the pancreas and retroperitoneal lymph node enlargement. His parents discontinued treatment. It is concluded that the rare causes of acute pancreatitis with poor response to conventional treatment should be considered, especially for patients with abdominal lymph node enlargement. Extranodal lymphoma should be considered, and lymph node biopsy should be performed as early as possible to confirm diagnosis. The prognosis of pancreatic lymphoma is associated with clinical stage and pathology.

Value of nasojejunal nutrition in the treatment of children with acute pancreatitis / 中华儿科杂志

<p><b>OBJECTIVE</b>To evaluate the feasibility and effectiveness of placement of nasojejunal feeding tube and nasojejunal nutrition feeding in children with acute pancreatitis.</p><p><b>METHOD</b>Twenty-two patients (of whom 13 had severe acute pancreatitis and 9 acute mild pancreatitis) who needed nutritional intervention were selected. They were from Department of Gastroenterology and Surgery during the years 2009 - 2012, and they were at high nutritional risk after STONGkid nutrition risk screening. The average age of them was 5 - 15 years (9.1 years ± 2.8 years). Assisted by endoscopy, the nasojejunal feeding tube was placed in 22 of 24 patients (in 2 cases of recurrent pancreatitis the tubes were placed again after extubation). Besides the use of regular fasting, antacids, inhibitors of trypsin secretion, and anti-infective treatment, 23 cases of all children got nasojejunal nutrition treatment as well. The outcome measures included the success rate, complications of endoscope-assisted nasojejunal tube placement. The children's tolerance and nutrition indicators (weight, blood lymphocytes count, erythrocytes count, serum albumin, serum creatinine, blood urea nitrogen) were observed before and after enteral nutrition therapy.</p><p><b>RESULT</b>Malnutrition evaluation was done 24 times before treatment among 22 patients, incidence of malnutrition was 33% in 22 cases. Placement of nasojejunal tube placement was attempted for a total of 24 times and was successful on first placement in 22 cases, in two cases the placement was successful on the second placement, so the success rate of the first attempt for placement was 92%. No significant complications were observed in any of the cases. Twenty-three of 24 cases were given standardized enteral nutrition (one case was not given enteral nutrition therapy but underwent ERCP due to obstructive jaundice). Twenty-two of 23 cases could tolerate enteral nutrition well, only 1 case was unable to tolerate enteral nutrition due to the pancreas schizophrenia, paralytic ileus. The treatment of jejunal feeding success rate was 96%. The feeding duration was 2 - 74 d (27.0 d ± 18.3 d). The adverse reactions include plugging of the tube in two cases, constipation in two cases, five cases had abdominal pain, diarrhea in 2 cases, vomiting in 2 cases and 1 case of jejunum retention. No case had nasopharynx ulcers, gastrointestinal perforation, gastrointestinal bleeding, re-feeding syndrome and infection etc. Blood erythrocytes count, serum creatinine, blood urea nitrogen were not significantly changed. Twenty of 23 cases were cured, 2 cases were improved and 1 case was unchanged.</p><p><b>CONCLUSION</b>Endoscope-assisted nasojejunal tube placement for children with acute pancreatitis is safe and feasible. Nasojejunal nutrition therapy is effective for acute pancreatitis patients who are at severe nutritional risk, especially for the improvement of the nutritional status of children.</p>

Management of Fournier's gangrene: a report of 23 cases / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the clinical characteristics and treatment of Fournier's gangrene.</p><p><b>METHODS</b>We retrospectively analyzed the clinical data of 23 cases of Fournier's gangrene, all with flare and pains in the scrotum and penis and different degrees of involvement of the scrotum, penis, perianal area, perineum, inguinal and lower extremities. The patients were treated by early debridement, incision-drainage, anti-infection and hyperbaric oxygen therapy, respectively. Scrotoplasty was performed for 11 of the cases, penile and scrotal dermatoplasty for 7, and penile amputation and urethral fistulation for 2 with penile necrosis. One of the cases underwent suprapubic cystostomy, and another 1 received colostomy.</p><p><b>RESULTS</b>Twenty of the patients were recovered and 3 (1 with diabetes and 1 with AIDS) died after surgery.</p><p><b>CONCLUSION</b>Fournier's gangrene is a fatal disease. Early diagnosis and timely surgical intervention are essential for the management of the disease.</p>

Clinical manifestations and endoscopic features of abdominal type Henoch purpura in children / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the clinical manifestations and endoscopic features of abdominal type Henoch purpura in children and improve the diagnostic level.</p><p><b>METHODS</b>Retrospective review was made on the clinical, endoscopic and histopathological features of 57 cases of children with Henoch purpura abdominal type who had been hospitalized from Jan. 2002 to May 2007. Upper gastrointestinal endoscopy was performed in all cases. All the cases had various digestive system symptoms without cutaneous purpura before endoscopy. Mucosal specimens were taken from sinus ventriculi and duodenum for histopathological analysis. Helicobacter pylori (H.pylori) infection was identified by rapid urease test and histology, and diagnosis of H.pylori infection was made when both were positive.</p><p><b>RESULTS</b>The common gastrointestinal symptoms of Henoch purpura were abdominal pain (46 cases), vomiting (32 cases), hematochezia (11 cases), diarrhea (4 cases) and abdominal distention (1 case). Three cases had arthralgia and joint swelling. The main laboratory findings were increased peripheral white blood cells (33 cases, 57.9%), 1/5 of cases had elevated C reactive protein (CRP), low serum albumin and seroperitoneum. Endoscopy demonstrated the damages to the mucosa, which varied from congestion, edema, petechia and ecchymosis (37 cases, 64.9%) to erosive and multiple ulcers (14 cases, 24.6%), granulation of mucosa in descendent duodenum (4 cases, 7.0%), and diffuse hemorrhage of mucosa (2 cases, 3.5%). The upper gastrointestinal endoscopy showed that the commonest and most serious position involved was the descendent duodenum (55 cases, 96.5%), followed by duodenal bulb (32 cases, 56.1%) and stomach (18 cases, 36.1%), esophagus was less involved (1 case, 1.8%). Histological manifestations showed swollen vascular endothelial cells of capillary vessels and small blood vessels, fibrotic necrosis of small vessels and bleeding, diffuse perivascular lymphocytic and neutrophilic infiltration and nuclear debris in mucosa and submucosa. Three cases (5.3%) were found infected with H. pylori. In 49 cases (86.0%) cutaneous purpura appeared 1 - 7 days after endoscopy. Eight cases had no cutaneous purpura until they left hospital. Two cases were lost to follow up and 6 cases (10.5%) remained free from cutaneous purpura were followed up until now (1 - 5 years).</p><p><b>CONCLUSION</b>Descending duodenum is the commonest and most serious position of upper gastrointestinal tract involved in Henoch purpura. Upper gastrointestinal endoscopy with the mucosal biopsy are useful for the early diagnosis of Henoch purpura.</p>

Gastric acid reflux and reflux esophagitis in children / 中华儿科杂志

<p><b>OBJECTIVE</b>To evaluate the significance of gastric acid reflux in children with reflux esophagitis (RE).</p><p><b>METHODS</b>Twenty-four-hour esophageal pH monitoring and gastroscopy were performed in 180 children suffered from vomiting. The relationship between RE, non-esophagitis (NE), non-erosive reflux disease (NERD) and gastroesophageal reflux (GER) was analyzed.</p><p><b>RESULTS</b>Sixty-five of the 180 patients were confirmed as having RE by endoscopy. Among them, the number of cases with grades I, II and III RE according to the diagnostic criteria by endoscopy was 37, 19 and 9 cases, respectively, while the other 115 cases were diagnosed as NE. The positive rate of acid reflux in RE group was 58.5% (38/65), while it was 42.6% (49/115) in NE group (chi(2) = 4.179, P < 0.05). All parameters of acid reflux in RE group except for the episode of reflux and the number of reflux longer than 5 min were significantly higher than those in NE group. None of the parameters of acid reflux except for Boix-Ochoa score in grade III RE patients was significantly higher than that in both grade II and grade I RE cases. However, the difference in acid reflux parameter between grade I and grade II RE patients had not reached statistical significance. The results also showed that the positive rate of pathological acid reflux was 48.3% (87/180). Among them, 38 cases were RE, while other 49 cases were NERD. The difference in acid reflux between these two groups was not significant.</p><p><b>CONCLUSIONS</b>Gastric acid reflux may play a major role in the development of RE in children, but may not be a sole pathogenic factor. The degree of acid reflux is not closely correspondent to the severity of RE. Acid reflux may not completely contribute to RE. Gastroscopy is very important for patients with reflux symptom.</p>

CD4+ and CD8+ T cells in gastric mucosa in children infected with Helicobacter pylori / 中华儿科杂志

<p><b>OBJECTIVE</b>To study the changes of gastric mucosal CD4(+) and CD8(+) T cells in Helicobacter pylori (Hp) infected children.</p><p><b>METHODS</b>Seventy nine patients with digestive tract symptoms were assessed by endoscopy, rapid urease test and histology. Forty four patients had Hp positive chronic superficial gastritis (Hp(+)CSG) and 35 patients had Hp negative chronic superficial gastritis (Hp(-)CSG). Gastric biopsy specimens were obtained from each patient. Peripheral blood samples were obtained from 33 patients (12 with Hp(+)CSG, 21 with Hp(-)CSG). Hp infection was identified by rapid urease test and histology. Hp infection was confirmed when a patient was positive for both of these tests. Four pieces of gastric antrum mucosal specimens were placed in Hank's balanced salt solution containing 1 mmol/L dithiothreitol (DTT) and 1 mmol/L ethylenediamine tetraacetic acid (EDTA). The specimens were treated with collagenase type I (120 U/ml) for three hours at 37 degrees C with agitation. The mononuclear cells were collected by removing undigested material and washed three times with RPMI 1640. Isolated gastic mononuclear cells were stained with CD3-FITC (fluorescein isothiocyanate), CD4-PE (R-phycoerthrin), CD8-PerCP (Peridinin-chlorophyll-alpha-protein) and measured by flow cytometry. Mucosal T lymphocytes were gated for the expression of CD3. Peripheral blood lymphocyte subsets were analysed by direct immunofluorescence.</p><p><b>RESULTS</b>The percentage of isolated gastric mononuclear cells within the CD3 gate were 3.26 +/- 1.98 in Hp(-)CSG, 4.37 +/- 1.97 in Hp(+)CSG. Relative CD4(+)(%), CD8(+)(%) and CD4(+)/CD8(+) of the CD3(+) cells respectively were 23.74 +/- 10.37, 47.04 +/- 12.00, 0.52 +/- 0.23 in Hp(-)CSG group, 40.28 +/- 11.35, 27.91 +/- 8.84, 1.55 +/- 0.52 in Hp(+)CSG group. CD4(+)(%), CD4(+)/CD8(+) in Hp(+)CSG group were significantly higher than those of Hp(-)CSG group and CD8(+)(%) was lower than those of Hp(-)CSG group (P < 0.01). There were no significant differences in peripheral blood T lymphocyte subsets between the two groups.</p><p><b>CONCLUSION</b>The difference of gastric T lymphocyte response between Hp(+)CSG and Hp(-)CSG in children indicated that the local cellular immune reaction may play a critical role in the pathogenesis of Hp infection.</p>

Effect of cadmium on growth and photosynthesis of tomato seedlings / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

A hydroponic experiment carried out to study the effect of five Cd levels on growth and photosynthesis of two tomato cultivars showed that the addition of 0.1 micromol/L Cd induced a slight increase in plant height of Hezuo 903 and the SPAD (the Soil-Plant Analyses Development) value of the 2 cultivars. However, at higher Cd levels, i.e., 1 and 10 micromol/L, root length and volume, plant height, and SPAD value were all significantly reduced. On an average of the 2 cultivars, exposure to 1 and 10 micromol/L Cd for 33 d reduced plant height by 18.9% and 46.4% and SPAD value by 11.2% and 31.6%, compared with control, respectively. Similarly, root length was reduced by 41.1% and 25.8% and root volume by 45.2% and 63.7%, respectively. The addition of Cd in the growth medium also had significant deleterious effect on net photosynthetic rate (Pn) and intracellular CO(2) concentration (Ci), with Pn being reduced by 27.2% and 62.1% at 1 micromol/L and 10 micromol/L Cd treatments compared to the control, respectively, while Ci increased correspondingly by 28.4% and 39.3%.

Protein and hordein fraction content in barley seeds as affected by sowing date and their relations to malting quality / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The effect of sowing date on grain protein, hordein fraction content and malting quality of two-rowed spring barley was investigated by using ten commercial cultivars with different grain protein content and the relationships among these traits were examined. The results showed that grain protein content and B hordein content increased as the sowing date postponed and were significantly affected by sowing date, while C and D hordein contents were less influenced by sowing date. There were significant differences in grain protein and hordein fraction content among the ten cultivars. The coefficient of variation of D hordein content was much larger than that of B and C hordein contents, suggesting its greater variation caused by different sowing dates. Beta-amylase activity and diastatic power were also significantly affected by sowing date, with malt extract being less affected. Significant differences in measured malt quality were found among the ten cultivars. Grain protein was significantly correlated with B hordein and malt extract positively and negatively, respectively. There was no significant correlation between beta-amylase activity or diastatic power and grain protein content. B hordein was negatively and significantly correlated with malt extract, but no significant correlations between C hordein, D hordein and malting quality traits.

Evaluation of clinical value of anorectal manometry for diagnosis of Hirschsprung's disease in neonate / 中华儿科杂志

<p><b>OBJECTIVE</b>Hirschsprung's disease (HD), one of the most common causes resulting in lower intestinal obstruction in children, is prone to be misdiagnosed or to be missed from diagnosis because of its atypical clinical symptoms and inconspicuous morphological findings by barium enema X-ray. Recently, this situation has been largely ameliorated by increased comprehension of anorectal kinetics and improvement of instrument for measurement of anorectal pressure. By now, anorectal manometry (ARMM) has been regarded as a routine means for functional assessment and diagnosis for anorectal disease. Nevertheless, the accuracy rate of diagnosis of HD in neonate by ARMM remains to be elucidated. In this study the clinical evaluation of anorectal manometry as an early diagnostic method for neonates with Hirschsprung's disease was appraised.</p><p><b>METHODS</b>Forty-two HD patients defined by pathological study of rectal tissue obtained via rectal mucous membrane biopsy or operation were recruited in this study. ARMM was performed in liquid transmission using PC polygraph high rate gastrointestinal dynamical detection system (PC Polygraf HR, CTD-synectics, Sweden), with 4-lumen catheter with which a small 5-cm-long balloon was connected at the terminus. All children were positioned on their left side or back during the procedure and the pressure transducers were placed in the mid-axillary line level. The results of ARMM performed before operation or biopsy were compared with the results of barium enema X-ray testing. The decrease of internal anal sphincter pressure as rectoanal inhibitory reflex (RAIR) was measured based on the fluctuation curve of pressure detected. HD was defined when no decrease of anal catheter pressure was detected after insufflation (RAIR positive), and suspected HD state was assessed with the presentation of incomplete relaxation or positive/negative results coexisted (RAIR abnormal) in canal.</p><p><b>RESULTS</b>Thirty patients (71.43%) were diagnosed as HD by ARMM including 18 patients who showed negative response to RAIR and 12 patients whose response was abnormal. While barium enema examinations were carried out in all the 45 patients, the results showed 5 HD patients and 14 suspected HD patients, giving an overall diagnostic accuracy of 45.24%. There were also 16 patients with positive ARMM response and negative barium enema findings together, and 5 patients with negative ARMM results and positive barium enema findings at the same time. There was a significant difference between the two diagnostic methods (chi(m)(2) = 4.76, P < 0.05).</p><p><b>CONCLUSION</b>Anorectal manometry seems to be a more reliable method for diagnosis of Hirschsprung's disease in neonate than barium enema X-ray. Because ARMM is a simple, safe and non-invasive method, it can be used as a screening test of choice in neonates with clinically suspected HD. But for final diagnosis, it is reasonable to combine ARMM with other diagnostic methods in HD patients.</p>

Prevalence of Helicobacter pylori resistant to clarithromycin, amoxicillin and metronidazole in children / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the prevalence of Helicobacter pylori (Hp) resistant to clarithromycin, amoxicillin and metronidazole in children.</p><p><b>METHODS</b>A total of 44 Hp clinical isolates were cultured from children patients with Hp gastritis and peptic ulcer who underwent gastroscopy during the period from October 2002 to November 2003 in Children's Hospital affiliated to Medical College of Zhejiang University. Nineteen of the patients were girls and 25 boys. The mean age was 8.66 +/- 2.66 years (3 approximately 14 years). The susceptibilities of the 44 Hp strains to clarithromycin, amoxicillin and metronidazole were tested by agar dilution test to determine minimum inhibitory concentration (MIC)(50), MIC(90) and resistance rates were also calculated.</p><p><b>RESULTS</b>Among the 44 Hp strains isolated from children, 8, 4, and 14 strains were resistant to clarithromycin, amoxicillin and metronidazole and the resistant rate was 18.2% (8/44), 9.1% (4/44) and 31.8% (14/44), with MICs for all strains ranging from < 0.125 microg/ml to 64 microg/ml, < 0.125 microg/ml to 64 microg/ml and < 0.125 microg/ml to 128 microg/ml, respectively. MIC(50) and MIC(90) of clarithromycin, amoxicillin and metronidazole were < 0.125 microg/ml, 8 microg/ml; < 0.125 microg/ml, 0.5 microg/ml; 0.5 microg/ml, 128 microg/ml, respectively. The prevalence of Hp resistant to clarithromycin was higher in children than that in adults, while to metronidazole was lower than that in adults with high MICs (32 - 128 microg/ml) and MIC(90) (128 microg/ml) of metronidazole for resistant isolates. Three multidrug resistant strains (6.8%) were simultaneously resistant to these three antibiotics.</p><p><b>CONCLUSIONS</b>The prevalence of Helicobacter pylori resistant to metronidazole was high in children, and the rate of resistance to clarithromycin was higher than that isolated from adults. Amoxicillin-resistant strains were now found, and multidrug resistant strains which resistant to the three antibiotics also emerged.</p>

Prevalence and mechanism of Helicobacter pylori resistance to clarithromycin in children / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the prevalence of Helicobacter pylori (H. pylori) resistance to clarithromycin (CLM) in children and to demonstrate the correlation of 23S rRNA gene mutation to clarithromycin resistance of Helicobacter pylori isolates.</p><p><b>METHODS</b>Totally 108 clinical strains of H. pylori were isolated from gastric biopsy specimens obtained from children who underwent endoscopy during the period from October 2002 to January 2004 in Children's Hospital Affiliated to Medical College of Zhejiang University. H. pylori was identified by morphology and biochemical tests after culture. Clarithromycin susceptibility of H. pylori isolates was determined by both E-test and two-fold agar dilution method. A strain was considered resistant when the MIC was defined as >or= 1 microg/ml. Genome DNAs of the 108 isolates were extracted and prepared for PCR to detect the corresponding gene in the V domain of the 23S rRNA. The amplified fragments were recognized and analyzed by restriction fragment length polymorphism (RFLP) when an additional restriction site is created by the mutation. The PCR products of all sensitive and resistant strains were digested with restriction enzyme BbsI and BsaI and were analyzed on a 1.5% agarose gel to discriminate different kinds of mutant genotype.</p><p><b>RESULTS</b>Sixteen of 108 isolates of H. pylori were resistant to clarithromycin by the agar dilution method and E-test method in clinical isolates from children, and the CLM resistance rate was 14.8% (16/108) with MICs ranging from 1 microg/ml to 128 microg/ml. Comparison of results of the two methods showed that these two methods were quite consistent in determination of susceptibility and resistance. The target fragment 425 bp in length containing 23S rRNA corresponding gene was successfully amplified. An A2144G mutation digested with BsaI was detected in 13 resistant isolates, but an A2143G mutation digested with BbsI in only 3 among all 16 clarithromycin resistant strains. None of the sensitive isolates was cleaved by either BsaI or BbsI enzyme, indicating that there was no mutation on them. It was also found that all the fragments from the resistant strains were not completely digested, and 425 bp uncut fragments were also visible and showed three bands indicating that they were heterozygotic strains with a mixture of wild-types and A-->G genotypes. In addition, in this study, no statistically significant difference between mutations at positions 2143 and 2144 with respect to the MIC was observed (r = 0.035, P > 0.05).</p><p><b>CONCLUSION</b>A high prevalence of clarithromycin-resistant H. pylori strains were detected among strains isolated from Chinese children studied. The 23S rRNA gene mutation at positions A2143G and A2144G plays an important role in clarithromycin resistance of H. pylori and A2144G mutation is the predominant finding among the resistant strains.</p>

Levels and mRNA expression of interleukin-8 in gastric and duodenal mucosa of children Infected with Helicobacter pylori / 中华儿科杂志

<p><b>OBJECTIVE</b>To determine the changes of IL-8 and IL-8 mRNA in gastric and duodenal mucosa of children with Hp infection, to study the effect of Helicobacter pylori (Hp) infection on the expression of IL-8 and IL-8 mRNA, and to evaluate its possible roles in the pathogenesis of gastric and duodenal mucosal inflammation in Hp related gastroduodenal diseases.</p><p><b>METHODS</b>Biopsy specimens were taken from the antral and duodenal mucosa on endoscopy in patients with or without Hp infection, which was diagnosed by urease test and Warshing-Starry staining. The expression of IL-8 in gastric and duodenal mucosa was determined by ELISA, the expression of IL-8 mRNA was determined by using RT-PCR.</p><p><b>RESULTS</b>Inflammation of gastric antral mucosa was more severe in Hp-positive group than in Hp-negative group. Active inflammation often existed on the basis of chronic inflammation in Hp-positive mucosa, and duodenal mucosa had mild chronic inflammation in Hp-positive group. Of 17 children who were not infected with Hp, 4 had pathologically normal gastric mucosa and had mild chronic gastritis, one child had an active chronic gastritis. Nineteen children were infected with Hp and all had chronic gastritis with signs of active inflammation. Gastric and duodenal mucosal IL-8 and IL-8 mRNA were higher in HP infected than in non infected children (IL-8: in gastric mucosa 24.66 - 177.77 pg/mg, 2.94 - 12.98 pg/mg, t = 12.34, P < 0.01; in duodenal mucosa: 4.28 - 47.76 pg/mg, 2.04 - 9.52 pg/mg, t = 7.18, P < 0.01. IL-8 mRNA: in gastric mucosa 2.37 - 4.99, 0.05 - 0.44, t = 29.29, P < 0.01; in duodenal mucosa 1.22 - 1.87, 0.01 - 0.23, t = 37.20, P < 0.01). Children with active chronic gastritis had higher interleukin-8 levels and IL-8 mRNA expression than those with inactive gastritis (IL-8 in gastric mucosa: 12.98 - 177.77 pg/mg, 2.04 - 10.43 pg/mg, t = 10.66, P < 0.01; in duodenal mucosa: 5.28 - 47.76 pg/mg, 3.19 - 8.14 pg/mg, t = 6.52, P < 0.01. IL-8 mRNA in gastric mucosa: 0.51 - 4.99, 0.01 - 0.44, t = 18.62, P < 0.01; in duodenal mucosa: 0.23 - 1.87, 0.01 - 0.20, t = 19.10, P < 0.01).</p><p><b>CONCLUSION</b>Higher expression of IL-8 and IL-8 mRNA was seen in Hp-positive gastric and duodenal mucosa and in active gastritis. IL-8 may play an important role in the local gastric and duodenal mucosal inflammatory infiltration with a large number of neutrophils when there is Hp infection.</p>