Expression and significance of TGF-β1, p38MAPK and BMP-7 protein in liver specimens of patients with alveolar hepatic echinococcosis / 中国血吸虫病防治杂志

Objective To detect the expression of transforming growth factor-β1 (TGF-β1), p38MAPK and bone morphogenetic protein-7 (BMP-7) protein in the liver specimens of patients with hepatic alveolar echinococcosis, and to investigate the potential role of TGF-β1, p38MAPK and BMP-7 protein in hepatic fibrosis caused by hepatic alveolar echinococcosis. Methods A total of 20 patients with hepatic alveolar echinococcosis were enrolled as study subjects, and hepatic specimens were sampled from the sites within 0.5 cm (Group A) and 0.5 to 1.5 cm from hepatic alveolar echinococcosis lesions (Group B), while normal liver specimens sampled from the sites 2 cm and greater from hepatic alveolar echinococcosis lesions served as controls (Group C). The fibrosis of liver specimens was pathological examined using HE and Masson staining, and the expression of TGF-β1, p38MAPK and BMP-7 protein was quantified in liver tissues using Western blotting. The associations of TGF-β1, p38MAPK and BMP-7 protein expression with hepatic fibrosis were assessed. Results HE staining showed the malaligned structure of hepatocytes and destruction of the structure of hepatic lobules at various degrees in liver specimens in groups A and B, with hepatocyte degeneration, atrophy and necrosis, hyperplasia of fibrous tissues and eosinophilic granulocyte infiltration seen, while no abnormal pathological alterations of liver tissues, normal hepatocyte structure and morphology and uniform size, no malaligned structure of hepatocytes, clear structure of hepatic lobules, no or mild hepatocyte degeneration or necrosis, and no eosinophilic granulocyte infiltration were seen in Group C. Masson staining showed that there was hyperplasia of multiple fibrous connective tissues in the liver portal areas in groups A and B, with fibrosis seen in hepatic lobules, while no obvious pathological changes were seen in Group C. There were significant differences seen in TGF-β1 (P < 0.001), p38MAPK (P < 0.01) and BMP-7 protein (P < 0.05) expression in liver tissues in groups A, B and C, and higher TGF-β1, p38MAPK and BMP-7 protein expression was quantified in groups A and B than in Group C (all P values < 0.05), while greater TGF-β1, p38MAPK and BMP-7 protein expression was detected in Group B than in Group C (all P values < 0.05). The expression of TGF-β1, p38MAPK and BMP-7 protein correlated positively with the severity of hepatic fibrosis (r = 0.866, 0.702 and 0.801, all P values < 0.05), and there were significant differences in TGF-β1 (F = 72.580, P < 0.01), p38MAPK (χ2 = 31.705, P < 0.01) and BMP-7 protein expression (χ2 = 48.388, P < 0.01) among liver tissues with different degrees of fibrosis. The TGF-β1 protein expression correlated positively with p38MAPK and BMP-7 protein expression (r = 0.607 and 0.702, both P values < 0.001), and the BMP-7 protein expression also correlated positively with p38MAPK protein expression (r = 0.456, P < 0.001). Conclusion The interaction among TGF-β1, p38MAPK and BMP-7 jointly participates in the development of hepatic fibrosis induced hepatic alveolar echinococcosis.

An overview of COVID-19 / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Pneumonia caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection emerged in Wuhan City, Hubei Province, China in December 2019. By Feb. 11, 2020, the World Health Organization (WHO) officially named the disease resulting from infection with SARS-CoV-2 as coronavirus disease 2019 (COVID-19). COVID-19 represents a spectrum of clinical manifestations that typically include fever, dry cough, and fatigue, often with pulmonary involvement. SARS-CoV-2 is highly contagious and most individuals within the population at large are susceptible to infection. Wild animal hosts and infected patients are currently the main sources of disease which is transmitted via respiratory droplets and direct contact. Since the outbreak, the Chinese government and scientific community have acted rapidly to identify the causative agent and promptly shared the viral gene sequence, and have carried out measures to contain the epidemic. Meanwhile, recent research has revealed critical aspects of SARS-CoV-2 biology and disease pathogenesis; other studies have focused on epidemiology, clinical features, diagnosis, management, as well as drug and vaccine development. This review aims to summarize the latest research findings and to provide expert consensus. We will also share ongoing efforts and experience in China, which may provide insight on how to contain the epidemic and improve our understanding of this emerging infectious disease, together with updated guidance for prevention, control, and critical management of this pandemic.

Present situation and progress of comprehensive treatments for hepatic alveolar echinococcosis / 中国血吸虫病防治杂志

Alveolar echinococcosis is a parasitic zoonosis that severely damages human health. Currently, radical surgical resection is the first choice for hepatic alveolar echinococcosis. For the advanced hepatic echinococcosis patients with refractory radical resection, the palliative surgery combined with chemotherapy, liver transplantation, drug therapy, and radiofrequency microwave ablation may provide comprehensive tools. This article reviews the current situation and progress of comprehensive treatments for hepatic alveolar echinococcosis.

Present situation and progress of comprehensive treatments for hepatic alveolar echinococcosis / 中国血吸虫病防治杂志

Alveolar echinococcosis is a parasitic zoonosis that severely damages human health. Currently, radical surgical resection is the first choice for hepatic alveolar echinococcosis. For the advanced hepatic echinococcosis patients with refractory radical resection, the palliative surgery combined with chemotherapy, liver transplantation, drug therapy, and radiofrequency microwave ablation may provide comprehensive tools. This article reviews the current situation and progress of comprehensive treatments for hepatic alveolar echinococcosis.

Influence of miR-122 on IFN-α treatment for HCV infection / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the influence of miR-122 on IFN-α treatment for HCV infection.</p><p><b>METHODS</b>Huh7.5.1 cells infected with HCV were treated with miR-122 mimics (20 nmol/L, 100 nmol/L, 400 nmol/L) and/or IFN-α (1000 IU/ml). The relative expression of HCV RNA was detected by real-time polymerase chain reaction (PCR). Huh7.5.1 cells were treated with different amounts of HCV (107 copies, 106 copies and 105 copies) and/or IFN-α (1000 IU/ml).</p><p><b>RESULTS</b>IFN-α suppressed the replication of HCV in a time-dependent manner, resulting in a ≊ 83% reduction of HCV at 48 h. MiR-122 mimics facilitated replication of HCV RNA in a dose-dependent manner (P<0.05). The antiviral effect of IFN-α was inverted to levels of miR-122 mimics (20 nmol/L, 100 nmol/L, 400 nmol/L), (73.3% ± 3.5% compared with 84% ± 4.5%, P>0.05; 64.67% ± 5.5% compared with 84% ± 4.5%, P>0.05; 56.33% ± 5.1% compared with 84% ± 4.5%, P<0.05). The antiviral effect of IFN-α was inverted to HCV load (105 copies group compared with 107 copies group, P<0.05).</p><p><b>CONCLUSION</b>MiR-122 facilitates replication of HCV RNA in the cell culture system; and the expression of miR-122 may partly counteract the anti-HCV effect of IFN-α.</p>

Effects of miR-122 on expression of hepatitis B virus proteins / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of miR-122 on the expression of hepatitis B virus (HBV) proteins.</p><p><b>METHODS</b>Anti-sense oligodeoxynucleotide (ASODN) of two different sequences against miR-122, anti-miR-122 and LNA-antimiR-122 (Locked nucleic acid), human miR -122 (hsa-miR-122), or the negative control anti-GFP were designed and synthesized, then transfected into HepG2.2.15 cells. After 24 h and 48 h, the levels of HBsAg and HBeAg in the supernatant were determined with a time-resolved immunofluorometric assay (TRFIA). HBV DNA in supernatant and miR-122 in cells were measured by quantitative real-time PCR.</p><p><b>RESULTS</b>After 48 h expressions of miR-122 in the LNA-antimiR-122 and anti-miR-122 groups were significantly suppressed and lower than those in the negative control (P<0.001), while the level of miR-122 in the hsa-miR-122 group was higher than that in the negative control (P<0.001). The expression of HBeAg and HBsAg in hsa-miR-122 group was lower than that in the negative control (P<0.01) 24 h and 48 h after transfection. The expression of HBeAg and HBsAg in the anti-miR-122 group and LNA-antimiR-122 group was significantly lower than that in negative control (P>0.001). The levels of viral DNA at both time-points in the various test groups were not significantly different from those of negative control group (P>0.05).</p><p><b>CONCLUSION</b>miR-122 may regulate HBV antigens and potentially affect the progress of pathogenesis, which might be the new targets for treatment of HBV infection.</p>

The relationship between systemic inflammatory response syndrome and severity of acute pancreatitis combined with plateau erythrocythemia / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the relationship between systemic inflammatory response syndrome(SIRS) and severity of acute pancreatitis combined with plateau erythrocythemia in the high altitude.</p><p><b>METHODS</b>A retrospective analysis on the clinical data which involved acute pancreatitis combined with plateau erythrocythemia (n = 40) and without plateau erythrocythemia (n = 40) admitted from September 2006 to September 2009 was conducted. According to the unified standards, these cases were divided into plateau erythrocythemia group and no plateau erythrocythemia group. The patients in plateau erythrocythemia group were further divided into severe group and mild group according to scores of APACHEII. The data was analyzed according to the patient with (or without) SIRS, SIRS's standard indicators, diagnostic parameter and relation of severity and duration of SIRS in acute pancreatitis combined with plateau erythrocythemia.</p><p><b>RESULTS</b>There was significantly discrepancy between plateau erythrocythemia group and no plateau erythrocythemia group not only in the incidence of patients who developed SIRS, but also in two items of patients fulfilling or not fulfilling diagnostic criteria of SIRS (P < 0.05). There was significant statistical difference in three items of diagnostic parameter of SIRS between plateau erythrocythemia group and no plateau erythrocythemia group (P < 0.05). Significant difference in two and three diagnostic parameter was found on severity of SIRS in acute pancreatitis combined with plateau erythrocythemia (P < 0.05). The more severity acute pancreatitis combined with plateau erythrocythemia was, the longer duration of SIRS was.</p><p><b>CONCLUSION</b>SIRS is highly correlated with the severity of SIRS in acute pancreatitis combined with plateau erythrocythemia in the high altitude.</p>

IQGAP1 is overexpressed in hepatocellular carcinoma and promotes cell proliferation by Akt activation

The scaffold protein IQGAP1 shows elevated levels in several cancer types, but its expression in hepatocellular carcinoma is unknown. We found that 58% of human hepatocellular carcinoma tissue samples had increased IQGAP1 expression compared to adjacent normal tissue. Overexpressing IQGAP1 raised the in vivo tumorigenicity of hepatocellular carcinoma cells, and forced overexpression of IQGAP1 in vitro stimulated cell proliferation. Cell growth was reduced by knockdown or mutation of IQGAP1, or by treatment of cells with a phosphotidylinositol 3-kinase inhibitor. To determine the mechanism by which IQGAP1 overexpression affected hepatocellular carcinoma cells, we confirmed its interaction in these cells with mammalian target of rapamycin (mTOR), a serine/threonine kinase that integrates signals about nutrient and energy status with downstream effectors that influence cell division. In addition, we discovered a new interaction involving IQGAP1, mTOR and Akt, which is a downstream target of mTOR. Akt phosphorylation on Ser-473, which is catalyzed by mTOR and required for Akt activation, increased with increasing amounts of IQGAP1, and decreased with IQGAP1 mutation. We hypothesize that IQGAP1 is a scaffold that facilitates mTOR and Akt interaction.

Treatment of fracture of multiple ribs with absorbable rib fixed nail and dacron flap in 12 patients / 中国骨伤

<p><b>OBJECTIVE</b>To introduce the methods of dacron flap application and its indications in treating fracture of multiple ribs, in order to reduce the incidence of the complications of fracture displacement.</p><p><b>METHODS</b>From September 2006 to March 2008, 12 patients with fracture of multiple ribs were treated with absorbable rib fixed nail and dacron flap. Included 8 males and 4 females,the age was from 22 to 51 years with an average of 38.2 years,the operative time was 2 hours to 3 days after injured. All the patients were closed injury and simultaneously accompanied with significant chest pain and chest tightness. 4 cases with dyspnea, blood in sputum and blood oxygen saturation decreased. The X-ray showed 3 cases of unilateral fracture and 9 cases of bilateral rib fractures and all cases accompanied with hemopneumothorax.</p><p><b>RESULTS</b>All patients were followed up from 2 to 26 months with an average of 8 months. All the fractures healed. According to clinical criteria, pain, breathing, ribs alignment etc. to observe the effect, 10 cases got excellent result, 1 case good, 1 case poor.</p><p><b>CONCLUSION</b>It is safe and effective to use absorbable rib fixed nails and dacron flap for treating fracture of multiple ribs and especially for the patients of osteoporosis, comminuted fracture or oblique fracture.</p>

A high-throughput diagnostic method for detecting pathogenic microbes / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To develop a high-throughput diagnostic method with suspension array technique for detecting pathogenic microbes.</p><p><b>METHODS</b>The probes and positive controls of 56 kinds of pathogenic microbes were designed, synthesized, and used to detect pathogenic microbes with suspension array technique.</p><p><b>RESULTS</b>Fluorescence signals of 56 positive controls were higher than those of the negative controls, and there was no cross-reaction between the probes and positive controls of different microbes.</p><p><b>CONCLUSION</b>Based on suspension array technique, the high-throughput diagnostic method may be useful in clinical detection of pathogenic microbes.</p>

Surveillance of viral contamination of invasive medical instruments in dentistry / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>To investigate the viral contamination of invasive medical instruments in dentistry and to provide health administrative institutions with surveillance data.</p><p><b>METHODS</b>Sterilized samples were randomly collected from the department of dentistry to detect HBV-DNA, HCV-RNA, HIV-RNA and HBsAg.</p><p><b>RESULTS</b>Of the invasive medical instruments that were sterilized with 2% glutaraldehyde, one of the samples was positive for HBV-DNA, and another sample was positive for HBsAg.</p><p><b>CONCLUSION</b>Though massive virus contamination of invasive medical instruments in dentistry has been reduced to a low level, the occurrence of contamination still remains.</p>

Construction and characterization of a cDNA library from human liver tissue with chronic hepatitis B / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>To construct a cDNA library from human liver tissue with chronic hepatitis B and check its quality for investigating the expression level of liver tissue infected by hepatitis B virus. This will then be used to find the relevant genes and interesting proteins associated with the development of hepatitis B.</p><p><b>METHODS</b>The total RNA from liver tissue with chronic hepatitis B was extracted and the mRNA was purified using TRIZOL method. Switching mechanism at 5' end of the RNA transcript (SMART) technique and CDS III/3' primer were used for first-strand cDNA synthesis. Long distance polymerase chain reaction (LD PCR) was then used to synthesize the double-strand cDNA that was then digested by Sfi I and fractionated by CHROMA SPIN-400 column. The longer than 0.4 kb cDNAs were collected and ligated to lambdaTriplEx2 vector. Then lambda phage packaging reaction and library amplification were performed. The qualities of both unamplified and amplified cDNA libraries were strictly checked by conventional titer determination. Fourteen plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector.</p><p><b>RESULTS</b>The titers of unamplifed and amplified libraries were 1.94 x 10(6) pfu/ml and 1.49 x 10(9) pfu/ml respectively. The percentages of recombinants from both libraries were 98.15% in unamplified library and 98.76% in amplified library. The lengths of the inserts were 1.23 kb in average, 1-2 kb in 64.29%, and 0.5-1.0 kb in 35.71%.</p><p><b>CONCLUSION</b>A high quality cDNA library from human liver tissue with chronic hepatitis B was successfully constructed.</p>

Construction and characterization of a cDNA library from human liver tissue of cirrhosis / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct a cDNA library from human liver tissue of cirrhosis.</p><p><b>METHODS</b>The total RNA from human liver tissue of cirrhosis was extracted using Trizol method, and the mRNA was purified using mRNA purification kit. SMART technique and CDSIII/3' primer were used for first-strand cDNA synthesis. Long distance PCR was then used to synthesize the double-strand cDNA that was then digested by proteinase K and Sfi I, and was fractionated by CHOMA SPIN-400 column. The cDNA fragments longer than 0.4 kb were collected and ligated to lambdaTripl Ex2 vector. Then lambda-phage packaging reaction and library amplification were performed. The qualities of both unamplified and amplified cDNA libraries was strictly checked by conventional titer determination. Eleven plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector.</p><p><b>RESULTS</b>The titers of unamplifed and amplified libraries were 1.03 x 10(6) pfu/ml and 1.36 x 10(9) pfu/ml respectively. The percentages of recombinants from both libraries were 97.24 % in unamplified library and 99.02 % in amplified library. The lengths of the inserts were 1.02 kb in average (36.36 % 1 approximately equals 2 kb and 63.64 % 0.5 approximately equals 1.0 kb).</p><p><b>CONCLUSION</b>A high quality cDNA library from human liver tissue of cirrhosis was constructed successfully, which can be used for screening and cloning new special genes associated with the occurrence of cirrhosis.</p>

Inhibition of HBV core antigen gene expression in human embryonic kidney cell line AD293 by plasmid-based RNAi / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To inhibit HBV core antigen gene expression with plasmid-based RNAi.</p><p><b>METHODS</b>The shRNA expression vector targeting HBV core antigen gene was designed and constructed. Human embryonic kidney cell line AD293 was co-transfected with HBcAg-EGFP fusion protein expression vector and shRNA expression vector transiently, and the cells without shRNA-transfection and with non-specific shRNA transfection were used as controls. Inhibitory effect of RNAi was detected by fluorescence-activated cell sorting (FACS) and real-time fluorescence quantificational RT-PCR.</p><p><b>RESULTS</b>HBV core antigen gene expression in AD293 was inhibited by shRNA, with the maximal inhibition rate of 76 % measured by FACS and of 63.1 % by real-time PCR.</p><p><b>CONCLUSION</b>Effective inhibition of HBV core antigen gene expression by plasmid-based RNAi provides an alternative for anti-HBV study in vitro, which has potential clinical application.</p>

Inhibition of binding peptides on replication of duck hepatitis B virus / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To study the inhibitory effect of binding peptides on duck hepatitis B virus (DHBV) replication in duck hepatocytes.</p><p><b>METHODS</b>Specific binding peptides to duck hepatitis B virus polymerase (DHBVP) were screened by phage display technology (PDT), then were sequenced and synthesized. Binding peptides were added into primary culture of duck hepatocytes infected with DHBV in vitro. DHBV-DNA in the cytoplasm, cell nucleus and medium supernatant was assayed over time.</p><p><b>RESULTS</b>Seven binding peptides were obtained after 3-round screening by PDT. Duck primary hepatocytes infected by DHBV were treated with above obtained binding peptides. The DHBV-DNA levels in medium supernatant and cytoplasm of duck hepatocytes treated with synthesized peptides (the 3rd and the 6th peptide) were significantly lower than those of control cells (P<0.05).</p><p><b>CONCLUSION</b>Specific binding peptides to DHBVP could inhibit the replication of DHBV.</p>

Clinical observations on patients with surgical treatment after heart valve prosthesis implantation / 中华全科医师杂志

Objective To evaluate the safety of surgical procedures for patients after heart valve prosthesis implantation.Methods Clinical data of 12 cases with heart valve prosthesis implantation undergone other surgical treatment from November 1996 to December 2005 were retrospectively analyzed.All the cases had routine oral warfarin with prothrombin time (PT) of 20.0—28.3 s averaged 23.5 s, international normalized ratio (INR) for prothrombin of 1.79—2.23 averaged 1.95 and heart functional class Ⅰ—Ⅲ.Among them,appendectomy was performed in three cases with acute appendicitis,reposition and repair in one with inguinal hernia,radical gastrectomy in two with gastric carcinoma,left hemicolectomy in one,cholecystectomy in three,left femoral head replacement in one,and bilateral high ligation and ablation of great saphenous vein in one.Elective surgical operation was performed in seven cases,and emergency operation in five.In those with elective surgery,warfarin was stopped 2—3 days before operation,while 5—10 mg vitamin K_1 was injected intramuscularly 6—8 hours before emergency surgery with preoperative median PT of 15.1 and 15.3 s and median INR of 1.24 and 1.30,respectively.In operation,5—10 mg vitamin K_1 were injected intravenously into the patients by drip depending on their bleeding on the surface of wound.ECG,blood pressure,hemoglobin and oxygen saturation were routinely monitored for all the cases intraoperatively and postoperatively.For the cases with heart function above class Ⅱ,fluid infusion was adjusted based on intubated central venous pressure,and for those with general anesthesia,analyses of blood gases and electrolyte were monitored routinely in operation.Results OPeration time averaged 20—160 rain in all the 12 patients,with blood loss 5—280 ml in average and without complications of massive hemorrhage,thrombosis and heart failure.Conclusions Surgical operation was safe for patients with heart valve prosthesis implantation,if preoperative PT and INR were adjusted to about 15 s and 1.30,respectively by cessation of warfarin or application of vitamin K_1,combined with careful manipulation and strengthened perioperative management.

DNAzymes in vitro inhibit the expression of hepatitis B virus genes / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate the inhibition effects of DNAzymes specific to Hepatitis B Virus(HBV) s gene and e gene on the expressions of Hepatitis B surface antigen(HBsAg) and Hepatitis B e antigen(HBeAg).</p><p><b>METHODS</b>DNAzymes DrzBS and DrzBC specific to HBV s gene ORF A157UG and e gene ORF A1816UG, respectively, were designed and synthesized. The inhibition effects of DrzBS or DrzBC on the expressions of HBV s and e genes were observed in 2.2.15 cells.</p><p><b>RESULTS</b>The expression of HBV s or e genes was dramatically depressed after 2.2.15 cells treated by DrzBS or DrzBC. The concentration for effective inhibition was within 0.1-2.5 micromol/L and the inhibition showed a dose dependence within that concentration range. The maximum inhibition was 94.2% and 91.8% for DrzBS and DrzBC, respectively. The inhibition was maintained for 72 hours. The efficiency of inhibiting HbsAg, HbeAg in 2.2.15 cells by DrzBS, DrzBC was higher than that by antisense oligonucleotides for the same target genes. The concentrations for effective inhibition of the DNAzymes were at least 10-fold lower compared with antisense oligonucleotides. Neither inhibition on the replication of HBV DNA nor toxicity to 2.2.15 cells was observed.</p><p><b>CONCLUSION</b>DrzBS and DrzBC can highly block the expressions of HBV s gene and e gene in 2.2.15 HBV cell model and are proved a specific and effective anti-HBV gene therapeutic means.</p>

Construction of HBV S gene recombinant and its immunity induced in mice / 浙江大学学报·医学版

OBJECTIVE: To investigate the feasibility of HBV DNA vaccines. METHODS: HBV S gene was obtained by PCR and the PCR product was cloned into pcDNA3. The recombinant was screened by antibiotics, identified by digestion and confirmed by sequencing. The plasmid was then transfected into mammalian cell COS-7 for transient expression. Then the recombinant was injected into mice and the immune responses induced in mice were investigated. RESULTS: The sequence of HBV S gene was correct and HBsAg could be detected in cells transfected with pcDNA3-S. After immunization, the positive rate in mice immunized with pcDNA3-S and pCMV-S was 70%(7/10) and 80% (8/10). The mean levels of anti-HBs were (32.14+/-13.79)mIU/ml and (28.50+/-11.87)mIU/ml respectively. There was no statistically significant difference between them P 0.05 . The mean levels of anti HBs in the control group and blank groups were both less than 10 mIU/ml. In mice immunized with pcDNA3-S and pCMV-S the results were (35.40+/-4.85)% and (38.20+/-7.69)% when E/T was 20:1, or (23.95+/-3.98)% and (24.55+/-3.59)% when E/T was 10:1, again showing no difference statistically (P>0.05). The specific CTL cytotoxicity rate of control and blank groups was both less than 5%. CONCLUSION: A specific humoral and cellular immune response can be induced in mice by intramuscular injection of pcDNA3-S.