Practical Value of Environment-friendly Sample Release Agent Combined with Ultrasound in the Preparation of Pathological Tissue Sections / 中山大学学报(医学科学版)

ObjectiveTo explore the practical value of environment-friendly sample release agent combined with ultrasound in the preparation of pathological tissue sections. MethodsFrom February 2013 to December 2022， 2 518 pathological specimens submitted by Foshan Municipal Hospital of Traditional Chinese Medicine were selected as the study objects. Two samples of the same specimen were randomly divided into two groups： the environment-friendly fast group， in which the pathological tissue sections were made by using the environment-friendly sample release agent combined with ultrasound； and the traditional group， in which formaldehyde， ethanol and xylene were used to make slices in the conventional way. The differences of hematoxylin （HE） staining effect， immunohistochemistry （IHC） staining effect and MDM2 gene detection result of atypical lipomatous tumor/highly differentiated liposarcoma （ALT/WDL） tissue sections between the two groups were compared. Results① The wax of the two groups' pathological tissues was dehydrated well and the tissue hardness was moderate. After HE staining， the sections of the two groups were intact， without cracks and tremor marks， and the contrast between nucleus and cytoplasm was appropriate， with good transparency， uniform staining， and no tissue loss. The excellent rate and score of HE staining in the environmental fast group were higher than those in the traditional group， but the difference was not statistically significant （χ2 = 3.125，P1 = 0.070；t = 0.965，P2 = 0.334）. ②After IHC staining of the two groups of sections， the positive location of the cells was accurate， the staining was specific and uniform， the staining intensity was moderate， the staining sensitivity was good， and there was no tissue loss. The excellent rate of IHC staining and the positive rate of IHC staining in the environmental fast group were lower than those in the traditional group， but the difference was not statistically significant （χ12 = 2.769，P1 = 0.092；χ22 = 0.800，P2 = 0.375）. ③The background and outline of the two groups of WDL tissue sections were clear， the staining was uniform， the cells were clear and visible， the nuclear boundary was clear， the hybridization signal was clear and bright under the background fluorescence， and there was no miscellaneous signal. The two groups of sections were hybridized successfully， and MDM2 showed positive amplification. The number of cells successfully hybridized in the environment-friendly fast group was lower than that in the traditional group， but the difference was not statistically significant （t = 1.414，P = 0.230）. ConclusionsThe tissue treatment method of using environment-friendly sample release agent combined with ultrasound can ensure the detection effect of HE staining， IHC staining and MDM2 gene detection of pathological tissue sections， and is more efficient and environment-friendly， suitable for promotion and use in hospitals at all levels.

Epidemiological characterization of sapovirus gastroenteritis aggregation in a kindergarten in Jinshan District， Shanghai / 上海预防医学

Objective:To analyze the epidemiological characteristics of an aggregational gastroenteritis and determine the genotypes of sapovirus， and to provide scientific basis for formulating effective control strategies. Methods:Unified case definition， active case search and descriptive epidemiology were used to analyze the epidemic. Feces or anal swabs of untreated students， teachers， canteen staff as well as canteen environment samples were collected. Norovirus and sapovirus nucleic acid tests were conducted by real-time fluorescent RT-PCR， and sapovirus nucleic acid was amplified by conventional RT-PCR. The gene region of capsid protein was analyzed by MEGA7.0 software and phylogenetic tree was constructed. Results:A total of 12 cases were reported in the epidemic， and the incidence rate was 44.44%. All reported cases， with vomiting symptoms， were found in the same class. The epidemic showed a point-based outbreak. The first case became the source of infection in class， and the epidemic lasted for 8 days. Real-time fluorescent RT-PCR assay confirmed that five children's feces were positive for sapovirus nucleic acid， and the first-episode children's feces were positive for sapovirus and GII norovirus nucleic acid. Sequence alignment result showed that the sapovirus strains belonged to GI.1 type with homologous genes. Conclusion:Based on the clinical manifestations， field epidemiological investigation and laboratory test results， we confirm that the first case of the epidemic in class is caused by GI.1 sapovirus infection. The epidemic is effectively controlled by comprehensive measures such as case isolation and disinfection.

Efficacy comparison between two kinds of vitrectomy in proliferative diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To compare the clinical effect of 23G and 25G+ vitrectomy for treatment of proliferative diabetic retinopathy (PDR).METHODS: A total of 128 PDR patients (195 eyes) requiring vitrectomy in our hospital from November 2013 to May 2016 were randomly divided into 25G+ group and 23G group, 64 cases (97 eyes) in 25G+ group and 64 cases (98 eyes) in 23G group.In 25G+ group, patients were treated by 25G+ vitrectomy.In 23G group, patients were treated by 23G vitrectomy.The visual acuity, as well as intraocular pressure (IOP), iatrogenic injury and complications in two groups were recorded before and 1d, 1wk, 1mo after treatment.The operation time was compared between two groups.RESULTS: The operation time in 25G+ group was lower than that in 23G group (P0.05).IOP in 25G+ group before surgery had no significant difference compared with those after surgery at 1d,1wk, and 1mo(P>0.05), which it was the same in 23G group.IOP of two groups in the same period had no significant difference (P>0.05).The incidence rate of iatrogenic injury in 25G+ group was 4.1%, which was significant lower than that of 23G group (13.3%) (P<0.05).The incidence rate of complication in 25G+ group was 3.1%, which was significant lower than that of 23G group (11.2%) (P<0.05).CONCLUSION: Both 23G and 25G+ vitrectomy are safe and effective treatment for PDR.However, 25G+ vitrectomy is the better choice for PDR for the shorter operation time, lower incidence rate of iatrogenic injury and fewer surgical complications.

Suprapubis-assisted umbilical laparoendoscopic mini-dual-site surgery for varicocele: a report of 80 cases / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the safety, effectiveness and feasibility of suprapubis-assisted umbilical laparoendoscopic mini-dual-site surgery (SAU-LEMDS) in the treatment of varicocele.</p><p><b>METHODS</b>This study included 80 varicocele patients aged 24 - 44 (mean 28.5 +/- 2.6) years, 25 cases of grade I, 45 cases of grade II and 10 cases of grade III, 58 cases in the left side, 6 in the right and 16 in both sides, and all with asthenospermia. The patients were treated by SAU-LEMDS under subarachnoid anesthesia combined with general anesthesia in a supine position with a head-down-feet-up slope of 15 degrees. Two 5 mm trocars were inserted bilaterally at the umbilical edge, one with a 5 mm 30 degrees laparoscope placed in it, and another into the abdominal cavity below the pubic hairline with a 5 mm laparoendoscopic clipper placed in it. The operation procedure was similar to that of standard laparoscopic ligation of spermatic veins, with reservation of the spermatic artery and double-ligation of spermatic veins. And the procedure was repeated for the contralateral lesion in the bilateral cases. Postoperative follow-up was conducted for the incidences of orchiatrophy and testicular hydrocele and changes of seminal parameters.</p><p><b>RESULTS</b>All the operations were successful, with the mean operation time of (10 +/- 5.0) min (range 8 to 25 min) for the unilateral cases and (18 +/- 6.5) min (range 15 to 30 min) for the bilateral cases, the mean blood loss of (1.5 +/- 0.5) ml (range 1 to 2 ml), and the mean postoperative hospital stay of (2 +/- 0.5) d (range 1.5 to 3 d). The patients were followed up for 6 -24 (12 +/- 2.5) months, which showed significant improvement in sperm motility as compared with the baseline ([28.53 +/- 5.21] vs [19.62 +/- 3.56]%, P < 0.05), with 28 cases (35.0%) restored to normal. Recurrence was found in 4 cases (5.0%). Testicular hydrocele occurred in 7 cases (8.75%), but orchiatrophy in none. The scars in the umbilicus and suprapubis were invisible because of the wrinkles and pubic hair.</p><p><b>CONCLUSION</b>SAU-LEMDS is safe, effective and feasible for the treatment of varicocele. It is superior to umbilical laparoendoscopic single-site surgery (U-LESS) for its less invasiveness, simpler operation, and better cosmetic appearance.</p>

Pathogeneses of erectile dysfunction after rectal cancer treatment / 中华男科学杂志

Rectal cancer is a common malignancy in the alimentary tract with an increasing incidence, the current treatments of which include surgery, radiotherapy, chemotherapy, and integrated comprehensive options. Sexual dysfunction, especially erectile dysfunction (ED), is one of the commonest complications in men after rectal cancer treatment and is generally attributed to the damage to the pelvic autonomic nerves. However, recent studies show that ED after rectal cancer treatment is a complex pathophysiological process associated with neurogenic, vasculogenic, and psychological factors. This article reviews the pathogeneses of ED after rectal cancer treatment in order to provide some theoretical evidence for its prevention and treatment.

Rapid detection of novel avian influenza virus subtype H7N9 by multiplex real-time RT-PCR / 病毒学报

In order to develop a rapid detection kit for novel avian influenza virus (AIV) subtype H7N9, two sets of specific primers and probes were designed based on the nucleotide sequences of hemagglutinin antigen (HA) and neuraminidase antigen (NA) of novel H7N9 virus (2013) available in GenBank to establish the method of TaqMan probe-based multiplex real-time RT-PCR for rapid detection of AIV subtype H7N9. The primer and probe of HA were for all H7 subtype AIVs, while the primer and probe of NA were only for novel N9 subtype AIVs. The results showed that this method had high sensitivity and specificity. This method was applicable to the testing of positive standard sample with a minimum concentration of 10 copies/microL; it not only distinguished H7 subtype from H1, H3, H5, H6, and H9 subtypes, but also distinguished novel N9 subtype from traditional N9 subtype. A total of 2700 samples from Zhuhai, China were tested by this method, and the results were as expected. For the advantages of sensitivity and specificity, the method holds promise for wide application.

Epidemiological features on 3 important viral diarrhea among children in Zhuhai during winter and spring / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore the epidemiological characteristics of viral diarrhea of norovirus (NV), sapovirus (SV) and astrovirus (AstV) among children in Zhuhai during winter and spring.</p><p><b>METHODS</b>Stool specimens were collected from children with viral diarrhea in Maternal and Child Health Hospital of Zhuhai from November 21, 2009 to April 3, 2010. Nucleic acid of NV, SV and AstV from negative specimens of rotavirus and adenovirus were detected by using Reverse transcription-polymerase chain reaction (RT-PCR), and the types of positive samples of NV were also classified at the same time.</p><p><b>RESULTS</b>The total detection rate of the three viruses is 21.49 percent, the highest detection rate is 29.05% in December 2009, the lowest detection rate is 12.20% in February 2010, 87.96% of positive specimens were from children patients aged from 0 to 30 months. The season detection rate of NV, SV and AstV are 14.70%, 2.75% and 4.04% respectively. There were significant differences of NV and SV detection rates in every month of the season, whereas the AstV detection rate was comparatively stable. The highest detection rate of NV is 34.09% in children patients aged from 12 to 18 months, the highest SV detection rate is 12.5% in children patients aged from 60 to 120 months, and the highest AstV detection rate is 16.67% in children patients aged from 24 to 30 months. All the NV were belong to G II genogroup.</p><p><b>CONCLUSIONS</b>NV is one of the main pathogens causing viral diarrhea among children in Zhuhai during winter and spring, SV and AstV are also important pathogens. So we should strengthen the monitoring of viral diarrhea caused by NV, SV and AstV in infants and young children.</p>

Thread-moxa in Zhuang folk medicine combined with acupuncture and external application drugs on AIDS patients with herpes zoster: a clinical observation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the efficacy of thread-moxa in Zhuang folk medicine (TM) combined with acupuncture and external application drugs for AIDS patients with herpes zoster (AHZ).</p><p><b>METHODS</b>A randomized, controlled clinical trial was conducted in 60 patients with AHZ. They were randomly assigned to the treatment group (treated with TM combined with acupuncture and Jingwanhong Scald Ointment) and the control group (treated with Famciclovir Tablet, nimesulide dispersible tablet, vitamin B1, ribavirin ointment). The treatment course was 14 days for both groups.The clinical efficacy, significant efficiency visual analog scale score (VAS), sleep quality score (QS), the postherpetic neuralgia rate in 1 year after treatment were observed.</p><p><b>RESULTS</b>The markedly effective rate was significantly higher in the treatment group than in the control group (86.7% vs. 53.3%, P < 0.01). There was no statistical difference in the total effective rate between the two groups (96.7% vs. 80.0%, P > 0.05). The post-treatment VAS, QS, the time for pain disappearance, skin repair, crusting, and 1-year postherpetic neuralgia incidence rate were significantly lower in the treatment group than in the control group (P < 0. 05, P < 0.01).</p><p><b>CONCLUSIONS</b>TM combined with acupuncture and Jingwanhong Scald Ointment was effective for treating AHZ patients. It relieved pain quickly, shortened their course of disease, and improved their quality of sleep.</p>

Experimental study of effect of low power laser on telomere length of cells / 生物医学工程学杂志

To investigate the effect of low power helium neon laser (He-Ne laser) on the telomere length of human fetal lung diploid fibroblast (2BS) cell, we used the laser (gamma = 632. 8 nm, P = 2 mW) to treat the young 2BS cells. Cell growth and proliferation was observed through MTT method after treating with low power laser. The relative telomere length of 2BS cells was detected by fluorescence real-time quantitative PCR (q-PCR). The results showed that the cells of the treated groups grew better than the untreated groups. The telomere DNA length of the old 2BS cells, treated by low power He-Ne laser when they were young, was longer than that of untreated group. The results of the present study indicated that the low power He-Ne laser might decrease shortening rate of telomere and delay the aging of cells. Therefore, this study provides the experimental basis for us to further investigate the effect of low power laser on cell aging at the gene level.

Molecular evolution of two lineages related to influenza B virus based on HA1 gene / 中华流行病学杂志

Objective To study the evolutionary characteristics and rules of two lineages on influenza B virus.Methods A total of 126 HA1 sequences of strains isolated during 1940 to 2012were downloaded from the GenBank.Time of the most recent common ancestor (TMRCA) and divergence of the two lineages were calculated based on the data from phylogenetic analysis of HA1gene,using Bayesian Markov Chain Monte Carlo (Bayesian-MCMC) and molecular clock method.Results The average amino acid variant ratios were ranged from 5.4％ to 10.2％ within the strains of influenza B virus isolated during 1978 to 2010.Compared with the Victoria-like strains,all Yamagatalike strains showed an amino acid deletion at 163th site,while some of them showing a deletion at position 166.HA1 gene of influenza B virus seemed not have been affected by positive selection except a few sites.The evolutionary average rate on HA1 gene was 2.138 × 10-3 substitutions/site/year (95％HPD:1.833 × 10-3-2.437 × 10-3 substitutions/site/year).The estimated dates for TMRCA of the two lineages of influenza B virus could be dated back to 1971 (95％ HPD:1969-1972),while the divergence times of the two lineages were 1973 (95％ HPD:1971-1974) and 1977 (95％ HPD:1975-1978) respectively.Conclusion Significant differences were found on HA1 gene between earlier and recent identified strains of Victoria and Yamagata lineage.Differences between the two lineages increased and showing the potential of dividing themselves into different subtypes in the future.More attention should be paid to these trends and the related epidemiological significance.

Study on the complete sequence of CA24 variant isolated during the acute hemorrhagic conjunctivitis outbreaks in Zhejiang province during 2002 to 2010 / 中华流行病学杂志

Objective To analyze the genetic characteristics of the complete sequence of coxsackievirus A24 variant(CA24v) isolated from acute hemorrhagic conjunctivitis (AHC) outbreaks in Zhejiang province during 2002 to 2010.Methods Complete sequences of CA24v epidemic strains isolated in different years were amplified under the RT-PCR assay,while the sequences of whole genome,VP1,and 3C region of Zhejiang strains were compared with epidemic strains isolated in other areas of China and abroad.Results The whole genome of Zhejiang CA24v strains isolated in 2002 and 2010 was 7456-7458 bp in length,encoding a polyglutamine protein which containing 2214 amino acid residues.There was a insertion with T on site 97 and 119 within 5' non-coding region between epidemic strain Zhejiang/08/10 and strains isolated in 2002.The rates of amino acid homology among Zhejiang/08/10 and other strains isolated since 2002 were between 94.7％ and 100.0％.Compared with the representative strains circulated within the recent 60 years,the largest average amino acid variations had been occurred on region 2A and 3A,with the ratios as 8.4％ and 7.3％ respectively.The smallest variation happened in region 3D,with the ratio only as 1.9％.The rates of stable amino acid variation on the whole genome between strains isolated since 1987 and 2002 were 38 and 20.P-distance within groups appeared that region 3C was more stable than VP1 of strains isolated in 2002-2010,and the 3D of early strain Jamaica/10628/87 might have had a nature of recombination but not observed on those epidemic strains in recent years.Conclusion Within the evolution of CA24v strains,the time course was more significant than the geographical differences.There had been sporadic epidemics of AHC caused by CA24v in Zhejiang province since 2002.

Effect of Yunnan baiyao powder and alcohol combined with VitB12 on exosmosis of vincristin / 中国实用护理杂志

Objective To observe the application effect of Yunnan baiyao powder and alcohol with VitB12 on exosmosis of vincristin. Methods The 40 patients with exosmosis of vincristin were randomly divided into the treatment group(22 cases)and the control group( 18 cases).The treatment group was given the Yunnan baiyao powder and alcohol with VitB12,the control group was given magnesium sulfate for hydropathic compress.The effective rate and red swelling and ulcer of the skin in the two groups were evaluated. Results The effective rate of the treatment group was higher than that of the control group,red swelling and the diameter of the ulcer of the skin on the fifth and seventh day were shorter than the control group. Conclusions The Yunnan baiyao powder and alcohol with VitB12 are effective in treatment of the exosmosis induced by vincristin.

Study on the genome sequence of measles viruses circulated in Zhejiang province during 1999 to 2011 / 中华流行病学杂志

Objective To study the genetic variations between measles vaccinc strain S191 and strains that circulated in Zhejiang province causing the epidemics during 1999 to 20 1 1.Methods Complete sequence of the nine Zhejiang measles strains were amplified by RT-PCR assay.Products were sequenced and the obtained sequences were aligned and analyzed with vaccine strains S191 and the major epidemic strains isolated in foreign countries.Results The homology of amino acid among the nine Zhcjiang strains were 98.77％-99.89％.The strains were not affected by positive selection and the variations on each gene were still in random drift.Compared to vaccine strain S191,there were 135 to 159 amino acid changes in Zhejiang measles virus,in which 113 points were common variable positions,resulting in mutations on five glycosylation sites.At the nucleotide level,the biggest differences between the Zhejiang strains and the vaccine strain S191 were found on N gcne,with the average divergent ratio as 5.5％,while the biggest one was P protein,in the amino acid level,with the average mutation rate as 7.7％.In addition,with the complete genome sequences,the genetic distance between Zhcjiang epidemic strains and vaccine strains was greater than the distances between epidenic strains of genotype D4,B3 and vaccine strains (t=-9.76,P＜0.05；t=-12.39,P＜0.05).Conclusion There were significant differcnccs found in the each of the genes between Zhejiang epidemic strains and the vaccine strain S191.The differences between the current vaccine strains and H genotypc epidemic strains were much larger than the differences between the vaccine and the forcign epidemic strains (genotype D4,B3).Therefore,wc should pay close attcntion to this trend,and to develop candidates for the dcvclopnent of vaccines,as early as possible.

Study on the mutations within the whole genome of influenza virus subtype A/H3N2 strains circulated in Zhejiang province from 1998 to 2009 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyze the consistency of evolution condition between HA gene and the whole genome of influenza virus subtype A/H3N2 strains isolated in Zhejiang province from 1998 to 2009, and to study the potential antigenic region on the whole genome.</p><p><b>METHODS</b>The sequences of whole genome of 19 Zhejiang influenza virus isolates circulated from 1998 to 2009, which conserved by influenza laboratory of Zhejiang Provincial Centre for Disease Prevention and Control, were amplified using RT-PCR assays. The obtained sequences were used to conduct phylogenetic analysis with 10 contemporaneous vaccine strains. Three methods, including comparison of the amino acid substitutions, calculation of the entropy value and the filtering of positive selection sites, were used to confirm the mutable sites on each gene.</p><p><b>RESULTS</b>The whole genome of influenza virus subtype A/H3N2 was 4466 amino acids in length, with 137 stable mutations. The 144, 158 aa of HA gene mutate four and three times respectively; 93, 143, 307, 370, 372 aa of NA gene and 450 aa of NP gene mutate twice, and there were 29% (12/41) and 77% (24/31) mutations of HA and NA genes occurred on the non-epitope regions respectively. Analysis of the entropy value suggest that many amino acid sites on the non-epitope regions were prone to mutation, including 3, 225, 361 aa of HA gene; 93, 143, 147, 150, 372 aa of NA gene; 113, 576, 586 aa of PB1 gene; 101,256, 382, 421, 437 aa of PA; 377, 450 aa of NP gene; 218 aa of M1 gene and 31 aa of M2 gene.</p><p><b>CONCLUSION</b>Based on the whole genome of influenza virus subtype A/H3N2 strains isolated in Zhejiang province in 1998 to 2009, there may be several unknown or new antigen sites existing on the non-epitope regions of HA and NA genes and parts of internal genes. The phylogenetic tree constructed based on the complete sequence was more comprehensive than on the HA gene to reflect the genetic relationship and law of evolution among the influenza virus strains.</p>

Myocardial regeneration and repair of infarcted heart by a new composition isolated from Geum japonicum / 中华心血管病杂志

<p><b>OBJECTIVES</b>To isolate the cardiogenic fraction, which can enhance cardiogenic differentiation of bone marrow-derived mesenchymal stem cells (MSC) from Geum japonicum. The therapeutic effect of the isolated cardiogenic fraction was further tested in a rat myocardial infarction (MI) model.</p><p><b>METHOD</b>Bioassay guided fractionation method was used for the isolation of the cardiogenic fraction, named as heart repair fraction (HRF). MI was induced by a permanent ligation of left anterior descending coronary artery. The rats exhibiting similarly decreased values of left ventricle ejection fraction (LVEF) and fraction shortening (LVFS) were used. The rats in test group (n = 10) were subject to HRF treatment (20 mg×kg(-1)×d(-1)) through gastric gavage daily for 4 weeks. Water alone (2 ml/d) was given through gastric gavage to rats in the control group (n = 10). The cardiac function was assessed by echocardiography at different time points. Masson trichrome staining was used for evaluation of the infarct size. Morphological and immunohistochemical studies were performed to investigate the HRF mediated myocardial regeneration.</p><p><b>RESULTS</b>LVEF (66.2% ± 6.9%) and LVFS (46.8% ± 5.8%) were significantly increased two weeks post HRF treatment compared with the values (LVEF: 55.7% ± 6.0% and LVFS: 36.4% ± 5.2%) in control rats (all P < 0.01). The improved heart function was further restored 4 weeks post HRF treatment (P < 0.01). Furthermore, the treatment of acute MI with this HRF significantly reduced the infarct size (19.0% ± 6.1%) compared with that (31.1% ± 8.6%) in control rats (P < 0.01). Substantial regeneration of cardiomyocytes in infarcted region of the HRF treated heart was also observed that replaced a considerable part of the infarcted heart tissues resulting in remarkable reduction of the infarct size.</p><p><b>CONCLUSION</b>The properties of this HRF isolated from Geum japonicum in stimulating substantial regeneration of myocardium in infarct region with consequently improved cardiac function appear to be new and represent a new approach for the treatment of MI.</p>

Transfection of pancreatic cancer cells BxPC-3 with recombinant plasmid pSilencer4.1-CMV neo-hTERT-siRNA and its silencing effects / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the transfection of pancreatic cancer cells BxPC-3 with recombinant plasmid pSilencer4.1-CMV neo-hTERT-siRNA and its silencing effects.</p><p><b>METHODS</b>Pancreatic cancer cells BxPC-3 transfected with recombinant plasmid pSilencer4.1-CMV neo-hTERT-siRNA were selected as target and divided into five groups: (1) T1 group (pSilencer4.1CMV neo-hTERT1-siRNA), (2) T2 group (pSilencer4.1CMV neo-hTERT2-siRNA), (3) Lipofectamine (Lipofectamine), (4) mismatch group(pSilence4.1CMV, as negative control), (5) cell control group(without transfection). The expression of hTERT mRNA was detected by RT-PCR. The viability of cells was measured by MTT method. The cell cycle and cell apoptosis was measured by flow cytometry. The expression of telomerase protein was measured by Western blot.</p><p><b>RESULTS</b>Compared with Lipofectamine group, negative control group and cell control group, the expression of hTERT-mRNA and telomerase protein in cells was downregulated significantly(P<0.05), the viability of BxPC-3 cells was decreased significantly (P<0.05), the ratio of cells in G0/G1 stage was increased, the ratio of cells in S stage and G2/M stage was decreased, and the ratio of apoptotic cells was increased significantly in T1 group and T2 group.</p><p><b>CONCLUSION</b>Recombinant plasmid T1 and T2 can downregulate the expression of hTERT mRNA and telomerase protein in BxPC-3 cells , and has good RNAi silencing effects. T1 and T2 can also inhibit the growth of BxPC-3 cells, block the cell cycle, promote the apoptosis of cells, and has anti-pancreatic cancer effects in vitro.</p>

Preparation of a 96-microwell plate DNA diagnostic chip for detection of foodborne bacteria and its application in an incident of food poisoning / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop a 96-microwell plate DNA diagnostic chip for simultaneous detection of 9 major foodborne bacteria.</p><p><b>METHODS</b>Type-specific PCR primers labeled with biotin and oligonucleotide probes were designed according to the conservative genes of 9 major foodborne bacteria Staphylococcus aureus, Salmonella spp., Escherichia coli O157:H7 (Stx1 and Stx2), Shigella spp., Listeria monocytogenes, Bacillus cereus, Yersinia enterocolitica, Vibrio cholerae and Vibrio parahaemolyticus. A one-tube multiplex PCR system for simultaneous amplification of these bacteria was established, and the DNA probes were spotted and immobilized in the wells of the plate in 5x5 array format. Stable hybridization system between PCR products and oligonucleotide probes in the microwell was established after condition optimization. Alkaline phosphatase-conjugated streptavidin and NBT/BCIP were used to detect the hybridized PCR products.</p><p><b>RESULTS</b>Twenty standard bacteria strains were used to validate the 96 microwell plate DNA diagnostic chip and highly specific and stable experiment results were obtained. Using this chip assay, the causal pathogen Staphylococcus aureus was identified within 12 h after the sampling from an incident of food poisoning, and the result was consistent with that obtained using conventional bacterial culture and biochemical identification.</p><p><b>CONCLUSION</b>The novel 96 microwell plate DNA diagnostic chip allows rapid, accurate, automated and high-throughput bacterial detection and is especially valuable for quick response to such public health emergencies as food poisoning.</p>

Traceability of the A/H3N2 virus borne influenza pandemic in Zhejiang province,1998 / 中华流行病学杂志

Objective To trace back to the influenza pandemic caused by A/H3N2 virus happened in Zhejiang province,1998.Methods The whole genome of three isolates related to Zhejiang influenza virus was amplified through RT-PCR,and the identified sequences were aligned with the sequences downloaded from GenBank of the H3N2 strains which were circulating in other regions during 1995 to 1998.The crossing HAI titers of the reference strains were measured by HAI test and antigenic ratios were calculated.Results The Phylogenetic tree,constructed based on HA sequence showed that the dominant strains A/Zhejiang/11/98 and A/Zhejiang/18/98 were significant different from the isolates circulated in other regions during 1995 to 1996 and the strains isolated in the mainland of China,in 1997.Although the A/Zhejiang/11/98 and A/Zhejiang/18/98 strains were distributed in the same cluster with A/Sydney/5/97,the two strains were closer to the epidemic strains isolated in Hong Kong and New York in the later part of 1997.Based on HAI,NA and MP genes,A/Zhejiang/18/98 seemed to be the closest one to the Hong Kong epidemic strains,and the genetic distances between A/Zhejiang/18/98 and New York strains were shorter than that with A/Sydney/5/97 based on PA,HA and NS genes.There were only 1-3 amino acid differences between A/Zhejiang/18/98 and Hong Kong or New York strains,whereas 7 amino acid differences with A/Sydney/5/97,in which three were located in the antigenic determinant regions.Data from the crossing HAI test showed that the antigenic ratio between A/Zhejiang/18/98 and A/Sydney/5/97 had reached 2.0,indicating the antigenic difference to a certain extent.Additionally,the onset of the influenza epidemic during 1997 to 1998 also suggested the possible route of transmission related to this H3N2 virus.Conclusion The influenza pandemic occurred in Zhejiang province in 1998 was possibly caused by the importation of a newly identified H3N2 influenza variant via Hong Kong and New York in late 1997.

Study on the genome sequence of influenza virus subtype A/H3N2 strains circulated in Zhejiang province during 1998 to 2009 / 中华流行病学杂志

Objective To analyze the relationship between influenza epidemic and genetic characteristic on the whole genome of influenza virus subtype A/H3N2 strains isolated in Zhejiang province during 1998 to 2009. Methods All of the eight genes from the 19 Zhejiang influenza virus isolates, circulated during 1998 to 2009, were amplified by RT-PCR and sequenced. The obtained sequences were aligned and analyzed with the vaccine strains being used in the last 10 years.Results The highest mutation happened within HA and NA genes and the amino acid divergent ratios were 13.98% and 10.00%. Amongst the six internal proteins, the amino acid divergent ratios of NP, M2 and NS1 were 6.43%, 6.19% and 3.48% respectively, and the others were lower than 3%.Other than the HA and NA genes, mutations were also observed on six internal genes of the strains isolated in those years when the influenza virus subtype A/H3N2 was widely circulating.Additionally, there had been an obvious genetic lag between vaccine strains recommended by WHO and the contemporary Zhejiang epidemic strains for many years. Conclusion Besides on HA and NA genes, surveillance programs should also be covered mutations regarding the internal genes of influenza virus subtype A/H3N2 strains, in order to provide important information for forecasting and warning of a new round of influenza epidemic.

One-step multiplex RT-PCR for rapid screening of type A, B and novel A (H1N1) influenza viruses / 南方医科大学学报

<p><b>OBJECTIVE</b>To developed a multiplex RT-PCR assay for simultaneous screening of type A, B and novel A (H1N1) influenza viruses.</p><p><b>METHODS</b>Two pairs of universal primers in were designed for amplifying the M gene and NS gene of type A and B influenza viruses, respectively. A pair of specific primers of HA gene was designed to detect novel A (H1N1) influenza virus. A one-step method was used to establish the multiplex RT-PCR system. A blinded experiment was carried out to validate the accuracy of this assay in comparison with the results of real-time fluorescence RT-PCR. The clinical practicability and efficacy of this assay was also evaluated.</p><p><b>RESULTS</b>The RT-PCR products were analyzed using agarose gel electrophoresis, which yielded distinct bands of the target fragments without non-specific reactions, suggesting the high efficiency and specificity of the multiplex RT-PCR. Blinded study of 50 samples demonstrated a concordance rate of 100%.</p><p><b>CONCLUSION</b>This multiplex RT-PCR assay allows one-step simultaneous detection of type A, B and novel A (H1N1) influenza viruses rapidly and accurately, and provides a valuable low-cost screening technique for influenza epidemic monitoring and early diagnosis.</p>