Prenatal diagnosis and outcome of pregnancy for women with high risks by screening of fetal free DNA from peripheral blood samples / 中华医学遗传学杂志

OBJECTIVE@#To analyze the results of prenatal diagnosis and outcome of pregnancy for women with a high risk for fetal aneuploidies.@*METHODS@#A total of 747 cases of prenatal diagnosis by amniocentesis due to high risks by non-invasive prenatal testing (NIPT) were selected from January 2015 to March 2022 in the Drum Tower Hospital Affiliated to Nanjing University Medical School. The amniotic fluid samples were subjected to chromosomal karyotyping and/or chromosomal microarray analysis. All cases were followed up by searching the birth information or telephone calls, and the results were recorded. 2 test or F test were used for comparing the difference between the groups.@*RESULTS@#Among the 747 pregnant women with a high risk by NIPT, 387 were true positives, and the overall positive predictive value (PPV) was 51.81%. The PPVs for trisomy 21 (T21), trisomy 18 (T18), trisomy 13 (T13) and sex chromosome aneuploidies (SCA) were 80.24% (199/248), 60% (48/80), 14% (7/50) and 38.97% (106/272), respectively. The PPV for T21 was significantly higher than T18 and T13 (χ2 = 85.216, P < 0.0001). The PPV for other chromosomal aneuploidies and copy number variations (CNVs) were 11.11% (5/45) and 40.74% (22/52), respectively. The PPV for increased X chromosomes was significantly higher than X chromosome decreases (64.29% vs. 22.22%, χ2 = 5.530, P < 0.05). The overall PPV for elder women (≥ 35 years old) was significantly higher than younger women (69.35% vs. 42.39%, χ2 = 49.440, P < 0.0001). For T21 and T18, the PPV of Z ≥ 10 group was significantly higher than that for 3 ≤ Z < 5 group or 5 ≤ Z < 10 group (P < 0.05). Among 52 cases with a high risk for CNVs, the PPV for the ≤ 5 Mb group was significantly higher than the 5 Mb < CNVs < 10 Mb or > 10 Mb groups (60% vs. 30%60% vs. 23.53%, P < 0.05). Among the 387 true positive cases, 322 had opted for induced labor, 53 had delivered with no abnormal growth and development, and 12 were lost during the follow-up.@*CONCLUSION@#The PPVs for common chromosomal aneuploidies are related to the age and Z value of the pregnant women, which were higher in the elder group and higher Z value group. In addition, the PPV is associated with high risk types. The PPV for T21 was higher than T18 and T13, and that for 45,X was lower than 47,XXX, 47,XYY or 47,XXY syndrome. NIPT therefore has relatively high PPVs for the identification of chromosomal CNVs.

Research advances in hepatitis B virus-associated glomerulonephritis / 临床肝胆病杂志

There is are large number of patients with chronic hepatitis B virus (HBV) infection. HBV not only damages the liver, but also involves the kidney. Hepatitis B virus-associated glomerulonephritis (HBV-GN) is secondary glomerulonephritis caused by HBV infection, and it is one of the most common extrahepatic complications of HBV infection. HBV-GN is mainly observed in children and young and middle-aged adults, with varying degrees of proteinuria as the main clinical manifestation, and it may be accompanied by hematuria and hypertension. Membranous nephropathy is the most common pathological type, followed by membrano-proliferative glomerulonephritis and IgA nephropathy. HBV-GN has an insidious onset and lacks characteristic symptoms and pathological manifestations, and thus it may be easily confused with various types of glomerulonephritis, which may lead to missed diagnosis and misdiagnosis. HBV-GN has a complex pathogenesis involving various links such as immune disorders, direct viral damage, and genetics, among which the theory of immune complex deposition has been widely recognized. In recent years, some important advances have been made in the research on the pathogenesis, diagnosis, and treatment of HBV-GN. This article summarizes the above issues, so as to provide a reference for clinical diagnosis and treatment.

Thoughts and practices of personnel training majoring in Chinese material medica based on the cultural self-confidence of Chinese medicine / 国际中医中药杂志

Guided by Chinese medicine culture,the strengthening of the cultural self-confidence of Chinese medicine is the cornerstone of the personnel training majoring in Chinese material medica.With the rapid development of modern medicine,the personnel training of the undergraduate students majoring in Chinese material medica in the universities of western medicine is facing several problems,such as lack of cultural identity,lack of professional confidence,unsteadiness of professional thinking,and changes of employment direction.For these problems,based on the main line "Solidify the cultural foundation of Chinese medicine,strengthen the self-confidence of Chinese material medica",our school selected the Chinese medicine cultural traveling,social practices and Chinese medicine employment guidance curriculum as the starting points,and carried out various of lively and vivid activities,which had positive impacts on the learning enthusiasm and professional love of the undergraduate students majoring in Chinese material medica.

Sonic Hedgehog promotes fibroblast-like synoviocytes proliferation via modulating the mitogen-activated protein kinase/extracellular signal-regulated kinase signaling pathway in rheumatoid arthritis / 中华风湿病学杂志

Objective To study the effect of mitogen-activated protein kinas/extracellular signalregulated kinase (MAPK/ERK) signaling pathway on cell proliferation modulated by Sonic Hedgehog (Shh) signaling in fibroblast-like synoviocytes (FLS) isolated from patients with active rheumatoid arthritis (RA).Methods The synovial tissue were collected by the synovial arthroscopic debridement or arthroscopic synovectomy of RA patients with active disease activity [disease activity score(DAS)28 ≥3.2].The RA-FLS were primarily cultured by the explanted culture,and then were treated with Shh agonist purmorphamine,inhibitor cyclopamine or MAPK/ERK signaling pathway inhibitor U0126,respectively.Western blotting was used to examine the phosphorylation level of ERK 1/2 (p-ERK1/2),which was the critical protein of MAPK/ERK signaling.The cell proliferation activity was detected using cell proliferation and cytotoxicity kit-8 (CCK8),and the cell proliferation rate was detected using a flow cytometry.Analysis of variance and Kruskal-Wallis H(K) test were used for statistical analysis.Results Compared with the control group,purmorphamine transiently increased p-ERK1/2 protein at the concentration of 1 μmol/L,and the peak activations of p-ERK1/2 took place at 15 min (P＜0.01).Cyclopamine and U0126 decreased the expression ofp-ERK1/2 protein (P＜0.01).After the RA-FLS treated with purmorphmine(1 μmol/L)for 48 hours,the cell proliferation activity was (114±4)％ and the percentage of S phase cells was (8.39±0.60)％,which was significantly higher than those of the control group (100±0)％ (P＜0.01) and (3.29±0.69)％ (P＜0.01).After treated with cyclopamine (10 μmol/L) for 48 hours,the cell proliferation activity of RA-FLS was (89±1)％ (P＜0.05) and the percentage of S phase cells was (1.53±0.22)％ (P＜0.05).When co-treated with purmorphamine (1 μmol/L) and U0126 (10 μmol/L),the cell proliferative activity was (89±2)％ (P＜0.05) and the percentage of S phase cells was(1.07±0.25)％(P＜ 0.05).Conclusion Shh may promote proliferation of RA-FLS via modulating MAPK/ERK signaling,which in turn contributes to hyperplasia of synovium and ultimately leading to RA.

Association of BCAS3 rs11653176 gene polymorphism with the occurrence of gout / 中华内分泌代谢杂志

Objective To explore the relationship between the polymorphisms of the primary gout predisposing gene (BCAS3) rs11653176 locus and the incidence of gout in Han Chinese men in coastal areas of Shandong Province. Methods One hundred and fifty-two cases of patients with gout remission,68 cases of acute stage,252 patients with hyperuricemia, and 280 healthy subjects, total males, were enrolled. Genotyping the rs11653176 locus of BCAS3 gene by TaqMan probe technique. The expression level of BCAS3 gene mRNA in each PBMC was measured by RT-qPCR. The levels of tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and interleukin-18(IL-18)in serum were measured by ELISA. Results The change of allele frequency of rs11653176 locus in BCAS3 gene was associated with gout(P<0.01). BCAS3 mRNA in patients with gout was significantly higher than that of healthy people and patients with hyperuricemia(P<0.01). In gout patients, the expression level of BCAS3 gene containing C allele was higher than that of T allele(mRNA,P<0.05). The inflammatory factors in the acute phase of gout were significantly higher than those in phases of remission and hyperuricemia(P<0. 01). Conclusion Changes in the allele frequency of BCAS3 alleles rs11653176(high C, low T)may contribute to the expression of this gene,and lead to gout. And the onset of gout is closely related to the production of inflammatory factors.

Practice on teaching of traditional Chinese medicine pharmaceutics experiments based on constructivism learning theory / 国际中医中药杂志

Based on the constructivism learning theory, we designed a new pharmaceutics of TCM experiment teaching course which focused on process of knowledge formation, including a self-designed experiments on the basis of primary experiment, a cooperative learning platform, and an formative evaluation system. The practice has showed that this new teaching method can arouse the participants' interest and initiative, help to enhance the teaching efficiency and performance.

The effect of probiotics in the treatment of patients with severe hepatitis and its influence on inflammatory cytokines / 中国基层医药

Objective To investigate effect of probiotics in the treatment of patients with severe hepatitis and its influence on inflammatory cytokines.Methods A retrospective analysis of the data of patients with severe hepati-tis were maded.There were 100 patients,which including 50 cases of the observation group and 50 cases of control group,the control group was given conventional treatment,the observation group was given Bacillus viable capsule (Zhengchangsheng) for 4 weeks,2 granule a time and three times a day.The clinical symptoms were observed before and after treatment,the blood was exsanguinated for detecting bowel flora,total bilirubin,liver function and inflamma-tory cytokines.Results The total effective rate of the observation group was 86%,which was significantly higher than 56% of the control group(χ2 =11.409,P <0.01).The enterobacter of the observation group after treatment was (6.71 ±1.30)lgn/g,which was lower than before treatment(t =3.602,P <0.05),the bifidobacterium,lactobacillus, bacteroides were (7.88 ±1.18)lgn/g,(8.08 ±1.65) lgn/g,(8.30 ±1.20)lgn/g,respectively,which were higher than before treatment(t =3.772,3.707,3.523,all P <0.05).The intestinal flora in the control group had no differ-ence before and after treatment.After treatment,the total bilirubin,ALT,plasma endotoxin of the observation group and the control group were (195.0 ±25.4)μmol/L and (307.6 ±40.5)μmol/L,(58.2 ±11.5)U/L and (90.2 ± 10.5)U/L,(0.18 ±0.06)EU/mL and (0.29 ±0.08)EU/mL,respectively,which were lower than before treatment (t =16.307,9.408,10.157 and 4.032,8.075,5.076,all P <0.01).The serum TNF -α,IL -6 levels of the obser-vation group were (106.2 ±12.5)pg/mL,(82.3 ±18.9)pg/mL,which were lower than before treatment(t =3.732, 4.017,all P <0.05),which of the control group were (130.5 ±20.6)pg/mL,(110.2 ±17.8)pg/mL,there was no difference before and after treatment.The IL -2,IL -10 levels of the observation group were (60.7 ±10.0)pg/mL, (31.7 ±6.6)pg/mL,which were significantly higher than before treatment(t =3.757,3.877,all P <0.05),which of the control group were (46.2 ±5.8)pg/mL,(23.4 ±5.1)pg/mL,which had no difference before and after treat-ment.The total bilirubin,ALT,plasma endotoxin,serum TNF -α,IL -6 levels of the observation group were lower than the control group(t =7.653,8.104,3.309,3.511,3.787,all P <0.05).The IL -2,IL -10 levels of the obser-vation group were higher than the control group(t =3.487,3.428,all P <0.05).Conclusion Probiotics can correct intestinal flora,reduce plasma endotoxin,inhibit inflammatory mediators,reduce inflammation of the liver damage, improve liver function and clinical symptoms and should be introduced in the treatment of severe hepatitis.

The clinical efficacy and the influence on T cell subsets of telbivudine in the treatment of HBeAg-positive chronic hepatitis B / 中国基层医药

Objective To investigate the clinical efficacy and the influence on T cell subsets of telbivudine in the treatment of HBeAg -positive chronic hepatitis B , guide clinical treatment .Methods 70 patients HBeAg -positive chronic hepatitis B in our hospital were randomly divided into the telbivudine group which were received telbi -vudine 600mg/d and lamivudine group which were received lamivudine 100mg/d,35 cases of each group ,both group had been treated for 48 weeks,the liver function (ALT,AST) were detected for making a statistics of ALT normaliza-tion,the serum were extracting for detecting HBV -DNA and making a statistics of HBV -DNA negative rate by PCR,the HbeAg were detected for making a statistics of HBeAg seroconversionby ECLIA ,T cell subsets ( CD3+,CD4+, CD8+,CD4+/CD8+) were detected by flow cytometry ,observed adverse events .Results The ALT,AST,HBV-DNA of the telbivudine group and lamivudine group after treatment were (36.04 ±7.62)U/L and (63.53 ±14.13)U/L, (34.26 ±8.51)U/L and (55.31 ±15.48)U/L,(2.22 ±0.73)copies/mL and (3.28 ±0.95)copies/mL,all indi-cators significantly lower than before treatment (t =24.826,18.564,7.495 and 17.413,13.331,5.442,all P<0.05),and the ALT,AST of the telbivudine group were significantly lower than the lamivudine group ( t=3.867, 3.774,3.498,all P<0.05).The ALT normalization,HBV-DNA negative rate,HBeAg seroconversion rate of the telbivudine group and lamivudine group were 97.14%and 74.29%,94.29%and 68.57%,62.86%and 37.14%, telbivudine group,the telbivudine group were significantly higher than that of lamivudine group (χ2 =7.467,7.652, 4.629,all P<0.05).The CD3+,CD4+,CD4+/CD8+of telbivudine group and lamivudine group after treatment were (66.3 ±3.8)%and (60.6 ±3.5)%,(38.8 ±5.4)% and (35.2 ±4.4)%,(1.33 ±0.16)% and (0.95 ±0.11)%,all indicaitons significantly higher than before treatment ( t =5.442,6.173,3.753 and 3.404,3.635, 3.222,all P <0.05),the telbivudine group were significantly higher than lamivudine group (t =3.473,3.207, 3.422,all P<0.05).Conclusion The telbivudine has better efficacy than the lamivudine in the treatment of HBeAg-positive chronic hepatitis B because of enhancing immune function ,can improves ALT normalization ,HBV-DNA negative rate and HBeAg seroconversion rate ,inhibit HBV replication .

The expression of miR-183 family in the pathogenesis and development of noise-induced deafness / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To detect the expression variation of microRNA-183 family in cochlea of animal model characterized by noise-induced deafness at various time points, and to explore the mechanisms responsible for noise-induced deafness.@*METHOD@#Fifty mice were randomly divided into 5 groups. In the experimental group, 40 mice were exposed to 2-4 kHz narrow band noise at 100 dB SPL 6h per day for 3 consecutive days. The rest 10 mice served as the control group without receiving any noise. Auditory brainsterm response (ABR) were examined at the 1st, 7th, 14th and 28th day compaired with the ABR before the experiment,to confirm noise lead to the permanent threshold shift. The pathological damage processes of hair cell were detected by the basilar membrane stretched techniques. Real-time reverse transcriptase-polymerase chain reaction (qRT-PCR) was apply to quantify the expression of microRNA183 family members. Statistical analysis was performed by the SPSS 17.0 software.@*RESULT@#The hearing of mice in the experimental group was significantly less than that in the control group. In the experimental group, the hearing of mice exposed to noise were markedly less when compared with the one exposure to null-noise. The hearing in the 1st day group was least among experimental groups, and the followed one was mice in the 7th day group. No statistical difference were observed between the 14th and 28th day groups (P > 0.05). The results of surface preparation showed that the outer hair cells were chaotic, deformational, and their number decreased is time-dependent. The missing of the outer hair cells occurred mainly in the first and second rows, while the inner hair cells were not pronouncedly missing. The qRT-PCR showed that the expressions of the three genes (miR-183/96/182)in the 1st day and 7th day group with exposure to noise were less than in the control group (P 0.05). The expressions rised in the 14th day experimental groups, whereas the 28th day group's expressions of the three genes decreased markedly which were more than that in the 1st day and 7th day group (P < 0.01).@*CONCLUSION@#After noise exposure for some time, the expressions of miRNA-183 family members have significant changes in animal model with noise-induced deafness, which indicated that the miRNA183 family members may play important roles in the pathogenesis and development of noise-induced deafness.

Subjective and Objective Assessment of Quality of Life for Coblation-Assisted Operation in Patients with Severe OSAHS / 天津医药

Objective To evaluate the subjective and objective improvement of quality of life in patients with low-temperature plasma-assisted modified uvulopalatopharyngoplasty (H-UPPP) combined with lymphoidectomy in radix lin-guae and coblation-channeling of the tongue(CCT) treating for severe obstructive sleep apnea hypopnea syndrome (OSAHS) , and compare the surgical efficacy with that of H-UPPP. Methods A total of 81patients with severe OSAHS were divided in-to treatment group (n=42) and control group (n=39). Treatment group underwent the low-temperature plasma-assisted H-UPPP combining lymphoidectomy in radix linguae and CCT,and control group underwent H-UPPP treatment. The apnea hy-popnea index (AHI), the lowest saturation of arterial oxygen (LSaO2) and Quebec sleep questionnaire (QSQ) and Epworth sleepiness scale (ESS) before operation and 6-month after operation were recorded and compared between two groups. Re-sults All of detection indicators were significantly improved after operation in treatment group (P＜0.01). The total surgical efficacy was significantly higher in treatment group than that of control group (83.3%vs 12.8%,χ2=40.225,P＜0.01). The to-tal improvement rate of five dimensions in QSQ such as daytime sleepiness symptoms (76.2%vs 25.6%), signs during the day (71.4%vs 10.2%) and night (54.8%vs 5.1%), emotion (54.8%vs 2.5%), ability of social intercourse (50.0%vs 2.5%) and the total score (57.1%vs 7.7%) after operation were significantly higher in treatment group than those of control group. The se-vere indexes of ESS decreased to 23.8% after operation in treatment group than those of control group (51.3%). Conclu-sion Low-temperature plasma-assisted H-UPPP combined with lymphoidectomy in radix linguae and CCT is effective in patients with severe OSAHS, which can also improve the quality of life postoperatively.

Diagnostic value of intraluminal contrast enhanced ultrasonography in evaluation of gastrointestinal stromal tumors / 生物医学工程学杂志

The aim of the present study was to investigate the diagnostic value of the contrast-enhanced ultrasonography for gastrointestinal stromal tumors (GISTs). The conventional transabdominal ultrasonography and contrast-enhanced ultrasonography using an intraluminal contrast agent were performed in 66 cases. The different diagnostic value between conventional transabdominal ultrasonography and contrast-enhanced hydrocolon ultrasonography was compared, including the depiction ratio and location accuracy. The morpholpgical features of GISTs were observed. The sensitivity of the contrast-enhanced hydrocolon ultrasonography in the depiction of the GISTs (57/66, 86.4%) was higher than that of the conventional transabdominal ultrasonography(33/66, 50%, P<0. 05). Furthermore, the accuracy of the contrast-enhanced ultrasonography in the location of the GISTs (52/57, 91. 2%) was higher than that of the conventional transabdominal ultrasonography (13/33, 39. 4%, P<0. 05). The GISTs with higher or lower risk have different ultrasonic features. The contrast-enhanced ultrasonography was a more valuable diagnostic method for GIST than the conventional transabdominal ultrasonography.

Expression of VCA genes of Epstein-Barr virus in Pichia pastoris and clinical application / 中华微生物学和免疫学杂志

Objective To analyze the application of recombinant VCA-BALF4(S) and VCA-BFRF3 proteins in serological diagnosis of nasopharyngeal carcinoma (NPC). Methods DNA extracted from the B95-8 cells was used as the templates in a polymerase chain reaction(PCR). VCA-BALF4 1008 bp (S) (aa 287-623) and VCA-BFRF3 531 bp(177 as) were generated and inserted into pPICZaA vector.The recombinant plasmids were transformed into GS115 yeast by electroporation. The yeast transformants in-duced by methanol expressed recombinant proteins. The recombinant proteins synthesized were coated to mi-croplate for detection of EBV-IgA antibody in NPC patients by ELISA. Results We have successfully se-cretly expressed the recombinant VCA-BALF4(S) and VCA-BFRF3 protein in the Pichia pastoris. The mo-lecular weight of products were approximately 37×10~3 and 18×10~3, respectively. The recombinant proteins VCA-BALF4(S) and VCA-BFRF3 in the culture supematant showed good immunoreactivity with IgA anti-bodies to EBV by Western blot. A novel ELISA was established using Pichia pastoris-expressed VCA-BALF4 (S) and VCA-BFRF3 proteins. Serum samples were collected from patients with NPC and healthy controls and using this ELISA tested. The sensitivity of VCA-BALF4(S) and VCA-BFRF3 tests in the NPC sera were 88.7% and 71.0%, whereas the specificity of normal individuals are 96.4% and 91.8% separately. Con-clusion The recombinant proteins BALF4 (S) and BFRF3 were highly secretly expressed in Pichia pastoris,the diagnostic value of two recombinant proteins in screening for NPC patients were primary evaluated and the valuable pPICZa-BALF4(S) yeast strain was obtained.

Influence of different anesthesia methods on postoperative changes in T-lymphocyte subsets and cytokines in the patients under want operation of stomach carcinoma / 中国基层医药

Objective To study the effects of different anesthesia methods on postoperative changes in T-lymphocyte subsets and IL-6、IL-10、TNF-α in the patients undergoing operation of stomach carcinoma.Methods 60 patients with stomach carcinoma were randomly divided into group I(general anaesthesia)and Ⅱ(general combined epidural anesthesia).The level of T-lymphocyte subsets and IL-6、IL-10、TNF-αt were respectively detected and compared before anesthesia,2 h after anesthesia,ld and 6 d after operation.Results CD3+ 、CD4+ 、CD4+/CD3+ decreased,but IL-6 increased at the end of 2 h after anesthesia,l d after operation compared with those before anesthesia(all P＜0.05).While IL-10、TNF-α had no statistical significance(all P＞0.05).At 6 d after operation,the values recovered to the level before anesthesia in two groups.Conclusion The immune function in patients with gastric cancer decreaseed after anesthesia,it was more serious in geteral anaesthesia than that of general combined epidural anesthesia.But anesthesia had little influence on inflammatory response.

Clinical features of 23 hepatolenticular degeneration patients with liver injure / 中国基层医药

Objective To realise the clinical features and to reduce the diagnostic errors of hepatolenticular degeneration patients with liver injure. Methods Clinical manifestations, results of las tests and admitting diagnosis of 23 hepatolenticular degeneration cases were reviewed. Results All cases showed the elevation of serum transami-nase and/or total bilirubin. 17 cases(73.9% ) had manifestation of liver malfunction while nervous system symptoms were merely found in 2 cases. The decreased ceruloplasmin and elevated 24 hour urinary cuprum were detected in 23 and 21 cases respectively. Positive K-F rings were observed in 17 of 23 patients. Misdiagnosis rate of admitting diagno-sis exceeded 43.5%. 2 cases died after treating with penicillamine. Conclusion Liver injure that is one of the com-mon clinical features results in the diagnostic errors, but the combination detection will improve the accuracy of hepa-tolenticular degeneration.

The relationship between HBV genotypes and anti-virus therapeutic efficacy of interferon / 中国基层医药

Objective To study the effects of Interferon(IFN)in chronic hepatitis B patients genotype B and C.Methods 20 cases were genotype B ,23 cases were genotype C,all the patients were treated with 5 million units of IFN-a-lb,im,qod,for 12 months,viral markers,liver function and adverse drug reactions were observed.Results There was no statistically significant difference between B,C genotype in the negative conversion rate of HBV-DNA (60.0% ,39.1%),the negative conversion rate of HBeAg(42.9% ,30.8%),anti-HBe seroconversion rate(35.7% ,23.1 %)and the rate ALT normalization(85.0% ,73.9%)(t = 1.86,0.69,0.68 ,0.79,P ＞ 0.05).Conclusion The results suggested that therapeutic efficacy of IFN-α-lb was not significantly different between B,C genotypes.

Antibacterial Activity of Cefepime Combined with Sulbactam for Carbapenem-resistant Acinetobacter spp / 中华医院感染学杂志

OBJECTIVE To study the in vitro activity of cefepime combined with sulbactam against carbapenem-resistant strains of Acinetobacter spp clinically isolated. METHODS The activity of each drug alone was determined against 24 isolates. Chequerborad synergy testing was then performed against all the isolates. RESULTS The percentage of the FIC index showed 33.3% synergism, 62.5% additive, 4.2% (1/24) indifference, and no antagonism tively. CONCLUSIONS Testing of cefepime and sulbactam combinations revealed synergism and additivity activity against some carbapenem-resistant strains(23/24) of Acinetobacter spp which could be likely to prove beneficial effect for the treatment of infections due to multidrug-resistant strains of Acinetobacter spp.

Expression of hepatitis B virus S gene in Pichia pastoris and application of the product for detection of anti-HBs antibody.

Antibody to hepatitis B surface antigen (HBsAb) is the important serological marker of the hepatitis B virus (HBV) infection. Conventionally, the hepatitis B surface antigen (HBsAg) obtained from the plasma of HBV carriers is used as the diagnostic antigen for detection of HBsAb. This blood-origin antigen has some disadvantages involved in high cost, over-elaborate preparation, risk of infection, et al. In an attempt to explore the suitable recombinant HBsAg for the diagnostic purpose, the HBV S gene was expressed in Pichia pastoris and the product was applied for detection of HBsAb. Hepatitis B virus S gene was inserted into the yeast vector and the expressed product was analyzed by sodium dodecyl sulphate polyacrolamide gel electrophoresis (SDS-PAGE), immunoblot, electronic microscope and enzyme linked immunosorbent assay (ELISA). The preparations of synthesized S protein were applied to detect HBsAb by sandwich ELISA. The S gene encoding the 226 amino acid of HBsAg carrying a hexa-histidine tag at C terminus was successfully expressed in Pichia pastoris. The His-Tagged S protein in this strain was expressed at a level of about 14.5 % of total cell protein. Immunoblot showed the recombinant HBsAg recognized by monoclonal HBsAb and there was no cross reaction between all proteins from the host and normal sera. HBsAb detection indicated that the sensitivity reached 10 mIu (micro international unit)/ml and the specificity was 100 % with HBsAb standard of National Center for Clinical Laboratories. A total of 293 random sera were assayed using recombinant S protein and a commercial HBsAb ELISA kit (produced by blood-origin HBsAg), 35 HBsAb positive sera and 258 HBsAb negative sera were examined. The same results were obtained with two different reagents and there was no significant difference in the value of S/CO between the two reagents. The recombinant HBV S protein with good immunoreactivity and specificity was successfully expressed in Pichia pastoris. The reagent for HBsAb detection prepared by Pichia pastoris-derived S protein showed high sensitivity and specificity for detection of HBsAb standard. And a good correlation was obtained between the reagent produced by recombinant S protein and commercial kit produced by blood-origin HBsAg in random samples.

Relationship between (TTTTA)n gene polymorphism in the apolipoprotein(a) 5' control region and atherosclerotic cerebral infarction in Han nationality of Hubei area / 中国组织工程研究

BACKGROUND: Apolipoprotein(a) [Apo(a)] plays some role in promoting the formation of atherosclerotic plaque, and contains pentanucleotide repeats(PNR), which has a key value in genic research and in forecast on the increased risk of early atherosclerosis cerebral infarction (ACI). But the relationship between ACI and Apo(a) PNR in different races needs to be further investigated.OBJECTIVE: To investigate the relationship between the Apo(a) PNR polymorphism and ACI.DESIGN: A case-control study based on the ACI patients and normal people of Han nationality in Hubei.SETTING: Department of Laboratory in a hospital of a university.PARTICIPANTS: From February 1998 to March 1999, 82 ACI patients (ACI group) and 153 healthy controls(control group) were selected from the Department of Neurology, Central South Hospital and Yatai Hospital of Wuhan University. All patients were Han nationality in Hubei without any relatives.METHODS: Serum lipoprotein(a) [Lp(a) ], total cholesterol(TC), high density lipoprotein cholesterol(HDL-c), low density lipoprotein cholesterol (LDL-c), triglyceride (TG), apolipoprotein A Ⅰ (ApoA Ⅰ) and apolipoprotein B (Apo B) were tested respectively. Meanwhile, the PNR in the 5' control region of the Apo(a) was detected with polymerase chain reaction(PCR) and high voltage polyacrylamide gels electrophoresis. The results were analyzed with controlled analysis.RESULTS: The levels of serum Lp(a) [ (239.9 ±225.4) mg/L], TC [(4.76±0.74) nmol/L], TG[(1.74±0.60) mmol/L] and LDL-C [ (2.84 ± 0. 63) mmol/L] were remarkably higher in ACI group than those in control group, which were(133.5 ±97.7) mg/L in serum Lp(a), (4. 29±0.72) mmol/L in TC, (1.05±0.52) mmol/L in TG and(2.84±0.63) mmol/L in LDL-C, however, the level of HDL-C[ (0.88± 0.17) mmol/L] was remarkably lower in ACI group than that in control group [ ( 1.03 ± 0. 35 ) mmol/L], the differences were all significant( t = 3.65to9.18, P ＜ 0.01) . The levels of ApoA Ⅰ [(1.13±0.15) mmol/L,(1.25±0.19) mmol/L] and ApoB[(0.93±0.12) mmol/L, (0.89± 0. 15 ) mmol/L] were no significant difference compared with those in control group. The duplicated frequency of the allele(TTTTA) 5(0. 098) in the ACI was remarkably higher than that in control(0. 026) (x2 = 5.62, P＜ 0. 05), The frequency of the allele(TTTTA) 9 in the ACI(0. 073) was remarkably lower than that in control (0. 213 ) (x2 = 7.83, P ＜ 0.01 ), The frequency of the allele(TTTTA) 5 was also associated with low TC and high Lp(a) levels.CONCLUSION: It is suggested that the Apo(a) PNR polymorphism are associated with the susceptibility to ACI, and involved in the development of ACI.

Expression of hepatitis B virus core gene in Pichia pastoris / 中国病理生理杂志

AIM: To study the expression of hepatitis B virus core gene in Pichia pastoris and to obtain high-level expressed recombinant HBcAg with good immunoreactivity and high specificity. METHODS: HBV core gene was amplified by PCR from plasmid pHBV1 which contained HBV whole DNA sequence. The PCR product was cloned into pGEM-T vector by TA cloning strategy. After confirmed by DNA sequence analysis, the gene of interest was inserted into the yeast expression vector pPIC9. The recombinant plasmid pPIC9-cAg was constructed and transformed into GS115 by electroporation. The recombinant yeast GS115 was induced by 0.5% methanol. The expressed product was analysed by SDS-PAGE,Western blot and ELISA. RESULTS: The restriction analysis and DNA sequence analysis proved that HBV core gene had already been cloned to yeast expression plasmid pPIC9. The expressed HBcAg existed in SDS-PAGE. Good immunoreactivity and high specificity of the recombinant HBcAg have been proved by ELISA and Western blot. The titre of the recombinant HBcAg in the cell lysate was 1∶ 12 800 . CONCLUSION: The recombinant plasmid pPIC9-cAg was successfully constructed. The recombinant HBcAg with good immunoreactivity and high specificity was successfully expressed in Pichia pastoris expression system and can be applied to further developing HBcAb immunoassay. [

Protective effects of ulinastatin on lung in patient undergoing lung resection after chemotherapy / 中华麻醉学杂志

Objective To investigate the protective effects of ulinastatin on the lungs in patients undergoing lung resection after chemotherapy for lung cancer.Methods Thirty ASA Ⅱ or Ⅲ patients with lung cancer(Ⅲ a)aged 54-71yr weighing 55-74 kg undergoing lung resection after chemotherapy were randomly divided into 2 groups(n=15 each):Ⅰ ulinastatin group received ulinastatin 10 000 U?kg~(-1) after induction of anesthesia and Ⅱ control group received normal saline instead of ulinastatin.The patients were premeditated with intramuscular pothidine 70 mg and scopolamine 0.3 mg or atropine 0.5 mg.Anesthesia was induced with midazolam 0.05 mg?kg~(-1),fentanyl 4 ?g?kg~(-1) propofol 0.5-1.0 mg?kg~(-1) and vecuronium 0.1 mg?kg~(-1) and maintained with 1%-2% isoflurane inhalation and vecuronium infusion at 0.06-0.08 mg?kg~(-1)?h~(-1).Blood samples were taken after induction of anesthesia(T_1,baseline),at 40 and 90 rain of one-lung ventilation(T_2,T_3) for determination of serum IL-6,IL-8,IL-10 and TNF-? concentrations.Lung specimen was taken from the operated lung at 90 min of one-lung ventilation for microscopic examination with light and electron microscopo.Results Serum IL-6,IL-8,IL-10 and TNF-? concentrations were all significantly increased during one-lung ventilation as compared to the baseline values at T_1 in both groups.Serum IL-6,IL-8 and TNF-? concentrations were significantly lower while Serum IL-10 concentration was significantly higher in ulinastatin group than in control group during one-lung ventilation(P＜0.05).The histopathologic changes of lung tissue were significantly less in group utinastatin than in group control.Conclusion Ulinastatin can effectively protect the lung in patients with lung cancer after chemotherapy by reducing systemic inflammatory response.