Safety and immunogenicity of a broad-spectrum mosaic vaccine as a booster dose against SARS-CoV-2 Omicron and other circulating variants

BACKGROUNDThe rising breakthrough infections caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants, especially Omicron and its sub-lineages, have raised an urgent need to develop broad-spectrum vaccines against coronavirus disease 2019 (COVID-19). We have developed a mosaic-type recombinant vaccine candidate, named NVSI-06-09, having immune potentials against a broad range of SARS-CoV-2 variants. METHODSAn ongoing randomized, double-blind, controlled phase 2 trial was conducted to evaluate the safety and immunogenicity of NVSI-06-09 as a booster dose in subjects aged 18 years and older from the United Arab Emirates (UAE), who had completed two or three doses of BBIBP-CorV vaccinations at least 6 months prior to the enrollment. The participants were randomly assigned with 1:1 to receive a booster dose of NVSI-06-09 or BBIBP-CorV. The primary outcomes were immunogenicity and safety against SARS-CoV-2 Omicron variant, and the exploratory outcome was cross-immunogenicity against other circulating strains. RESULTSA total of 516 participants received booster vaccination. Interim results showed a similar safety profile between NVSI-06-09 and BBIBP-CorV booster groups, with low incidence of adverse reactions of grade 1 or 2. For immunogenicity, by day 14 after the booster vaccination, the fold rises in neutralizing antibody geometric mean titers (GMTs) from baseline level elicited by NVSI-06-09 were remarkably higher than those by BBIBP-CorV against the prototype strain (19.67 vs 4.47-fold), Omicron BA.1.1 (42.35 vs 3.78-fold), BA.2 (25.09 vs 2.91-fold), BA.4 (22.42 vs 2.69-fold), and BA.5 variants (27.06 vs 4.73-fold). Similarly, the neutralizing GMTs boosted by NVSI-06-09 against Beta and Delta variants were also 6.60-fold and 7.17-fold higher than those boosted by BBIBP-CorV. CONCLUSIONSA booster dose of NVSI-06-09 was well-tolerated and elicited broad-spectrum neutralizing responses against SARS-CoV-2 prototype strain and immune-evasive variants, including Omicron and its sub-lineages. The immunogenicity of NVSI-06-09 as a booster vaccine was superior to that of BBIBP-CorV. (Funded by LIBP and BIBP of Sinopharm; ClinicalTrials.gov number, NCT05293548).

Safety and immunogenicity of a hybrid-type vaccine booster in BBIBP-CorV recipients: a randomized controlled phase 2 trial

The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants with immune escape ability raises the urgent need for developing cross-neutralizing vaccines against the virus. NVSI-06-08 is a potential broad-spectrum recombinant COVID-19 vaccine that integrates the antigens from multiple SARS-CoV-2 strains into a single immunogen. Here, we evaluated the safety and immunogenicity of NVSI-06-08 as a heterologous booster dose in adults previously vaccinated with the inactivated vaccine BBIBP-CorV in a randomized, double-blind, controlled, phase 2 trial conducted in the United Arab Emirates (NCT05069129). Three groups of healthy adults over 18 years of age (600 participants per group) who had administered two doses of BBIBP-CorV 4-6-month, 7-9-month and >9-month earlier, respectively, were vaccinated with either a homologous booster of BBIBP-CorV or a heterologous booster of NVSI-06-08. The primary outcome was immunogenicity and safety of booster vaccinations. The exploratory outcome was cross-reactive immunogenicity against multiple SARS-CoV-2 variants of concerns (VOCs). The incidence of adverse reactions was low in both booster vaccinations, and the overall safety profile of heterologous boost was quite similar to that of homologous boost. Heterologous NVSI-06-08 booster was immunogenically superior to homologous booster of BBIBP-CorV. Both Neutralizing and IgG antibodies elicited by NVSI-06-08 booster were significantly higher than by the booster of BBIBP-CorV against not only SARS-CoV-2 prototype strain but also multiple VOCs. Especially, the neutralizing activity induced by NVSI-06-08 booster against the immune-evasive Beta variant was no less than that against the prototype strain, and a considerable level of neutralizing antibodies against Omicron (GMT: 367.67; 95%CI, 295.50-457.47) was induced by heterologous booster, which was substantially higher than that boosted by BBIBP-CorV (GMT: 45.03; 95%CI, 36.37-55.74). Our findings showed that NVSI-06-08 was safe and immunogenic as a booster dose following two doses of BBIBP-CorV, which was immunogenically superior to homologous boost with another dose of BBIBP-CorV. Our study also indicated that the design of hybrid antigen may provide an effective strategy for broad-spectrum vaccine developments.

Safety and immunogenicity of a heterologous boost with a recombinant vaccine, NVSI-06-07, in the inactivated vaccine recipients from UAE: a phase 2 randomised, double-blinded, controlled clinical trial

BackgroundThe increased coronavirus disease 2019 (COVID-19) breakthrough cases pose the need of booster vaccinations. In this study, we reported the safety and immunogenicity of a heterologous boost with a recombinant COVID-19 vaccine (CHO cells), named NVSI-06-07, as a third dose in participants who have previously received two doses of the inactivated vaccine (BBIBP-CorV) at pre-specified time intervals. Using homologous boost with BBIBP-CorV as control, the safety and immunogenicity of the heterologous boost with NVSI-06-07 against various SARS-CoV-2 strains, including Omicron, were characterized. MethodsThis study is a single-center, randomised, double-blinded, controlled phase 2 trial for heterologous boost of NVSI-06-07 in BBIBP-CorV recipients from the United Arab Emirates (UAE). Healthy adults (aged [≥]18 years) were enrolled and grouped by the specified prior vaccination interval of BBIBP-CorV, i.e., 1-3 months, 4-6 months or [≥]6 months, respectively, with 600 individuals per group. For each group, participants were randomly assigned at 1:1 ratio to receive either a heterologous boost of NVSI-06-07 or a homologous booster dose of BBIBP-CorV. The primary outcome was to comparatively assess the immunogenicity between heterologous and homologous boosts at 14 and 28 days post-boosting immunization, by evaluation of the geometric mean titers (GMTs) of IgG and neutralizing antibodies as well as the corresponding seroconversion rate ([≥]4-fold rise in antibody titers). The secondary outcomes were the safety profile of the boosting strategies within 30 days post vaccination. The exploratory outcome was the immune efficacy against Omicron and other variants of concern (VOCs) of SARS-CoV-2. This trial is registered with ClinicalTrials.gov, NCT05033847. FindingsA total of 1800 individuals who have received two doses of BBIBP-CorV were enrolled, of which 899 participants received a heterologous boost of NVSI-06-07 and 901 received a homologous boost for comparison. No vaccine-related serious adverse event (SAE) and no adverse events of special interest (AESI) were reported. 184 (20{middle dot}47%) participants in the heterologous boost groups and 177 (19{middle dot}64%) in the homologous boost groups reported at least one adverse reaction within 30 days. Most of the local and systemic adverse reactions reported were grades 1 (mild) or 2 (moderate), and there was no significant difference in the overall safety between heterologous and homologous boosts. Immunogenicity assays showed that the seroconversion rates in neutralizing antibodies against prototype SARS-CoV-2 elicited by heterologous boost were 89{middle dot}96% - 97{middle dot}52% on day 28 post-boosting vaccination, which was much higher than what was induced by homologous boost (36{middle dot}80% - 81{middle dot}75%). Similarly, in heterologous NVSI-06-07 booster groups, the neutralizing geometric mean titers (GMTs) against the prototype strain increased by 21{middle dot}01 - 63{middle dot}85 folds from baseline to 28 days post-boosting vaccination, whereas only 4{middle dot}20 - 16{middle dot}78 folds of increases were observed in homologous BBIBP-CorV booster group. For Omicron variant, the neutralizing antibody GMT elicited by the homologous boost of BBIBP-CorV was 37{middle dot}91 (95%CI, 30{middle dot}35-47{middle dot}35), however, a significantly higher level of neutralizing antibodies with GMT 292{middle dot}53 (95%CI, 222{middle dot}81-384{middle dot}07) was induced by the heterologous boost of NVSI-06-07, suggesting that it may serve as an effective boosting strategy combating the pandemic of Omicron. The similar results were obtained for other VOCs, including Alpha, Beta and Delta, in which the neutralizing response elicited by the heterologous boost was also significantly greater than that of the homologous boost. In the participants primed with BBIBP-CorV over 6 months, the largest increase in the neutralizing GMTs was obtained both in the heterologous and homologous boost groups, and thus the booster vaccination with over 6 months intervals was optimal. InterpretationOur findings indicated that the heterologous boost with NVSI-06-07 was safe, well-tolerated and immunogenic in adults primed with a full regimen of BBIBP-CorV. Compared to homologous boost with a third dose of BBIBP-CorV, incremental increases in immune responses were achieved by the heterologous boost with NVSI-06-07 against SARS-CoV-2 prototype strain, Omicron variant, and other VOCs. The heterologous BBIBP-CorV/NVSI-06-07 prime-boosting vaccination may be valuable in preventing the pandemic of Omicron. The optimal booster strategy was the heterologous boost with NVSI-06-07 over 6 months after a priming with two doses of BBIBP-CorV. Research in contextO_ST_ABSEvidence before this studyC_ST_ABSWe searched PubMed for clinical trials or prospective/cohort studies involving heterologous booster vaccination in non-immunocompromised population published up to Dec 25, 2021, using the term "(COVID) AND (vaccin*) AND (clinical trial OR cohort OR prospective) AND (heterologous) AND (booster OR prime-boost OR third dose)" with no language restrictions. Nine studies of heterologous prime-boost vaccinations with adenovirus-vector vaccines (ChAdOx1 nCov-19, Oxford-AstraZeneca, Ad26.COV2.S, Janssen) and mRNA vaccines (BNT162b2, Pfizer-BioNtech; mRNA1273, Moderna) were identified. The adenovirus-vector and mRNA heterologous prime-boost vaccination was found to be well tolerated and immunogenic. In individuals primed with adenovirus-vector vaccine, mRNA booster vaccination led to greater immune response than homologous boost. However, varied results were obtained on whether heterologous boost was immunogenically superior to the homologous mRNA prime-boost vaccination. Besides that, A preprint trial in population previously immunized with inactivated vaccines (CoronaVac, Sinovac Biotech) showed that the heterologous boost with adenovirus-vector vaccine (Convidecia, CanSino Biologicals) was safe and induced higher level of live-virus neutralizing antibodies than by the homogeneous boost. A pilot study reported that boosting with BNT162b2 in individuals primed with two doses of inactivated vaccines (BBIBP-CorV) was significantly more immunogenic than homologous vaccination with two-dose of BNT162b2. In addition, a preprint paper demonstrated that heterologous boost of ZF2001, a recombinant protein subunit vaccine, after CoronaVac or BBIBP-CorV vaccination potently improved the immunogenicity. But only a small size of samples was tested in this study and the live-virus neutralization was not detected. Till now, it is still lacking a formal clinical trial to evaluate the immunogenicity and safety of the heterologous prime-boost vaccination with an inactivated vaccine followed by a recombinant protein subunit-based vaccine. Added value of this studyTo our knowledge, this is the first reported result of a large-scale randomised, controlled clinical trial of heterologous prime-boost vaccination with an inactivated vaccine followed by a recombinant protein subunit vaccine. This trial demonstrated that the heterologous prime-booster vaccination with BBIBP-CorV/NVSI-06-07 is safe and immunogenic. Its immunoreactivity is similar to that of homologous vaccination with BBIBP-CorV. Compared to homologous boost, heterologous boost with NVSI-06-07 in BBIBP-CorV recipients elicited significantly higher immunogenicity not only against the SARS-CoV-2 prototype strain but also against Omicron and other variants of concern (VOCs). Implications of all the available evidenceBooster vaccination is considered an effective strategy to improve the protection efficacy of COVID-19 vaccines and control the epidemic waves of SARS-CoV-2. Data from our trial suggested that the booster vaccination of NVSI-06-07 in BBIBP-CorV recipients significantly improved the immune responses against various SARS-CoV-2 strains, including Omicron. Due to no Omicron-specific vaccine available currently, the BBIBP-CorV/NVSI-06-07 heterologous prime-boost might serve as an effective strategy combating Omicron variant. Besides that, BBIBP-CorV has been widely inoculated in population, and thus further boosting vaccination with NVSI-06-07 is valuable in preventing the COVID-19 pandemic. But further studies are needed to assess the long-term protection of BBIBP-CorV/NVSI-06-07 prime-booster vaccination.

Review of risk evaluation scores for benign end stage liver diseases recipients / 中华外科杂志

Liver transplant is an unreplaceable method for benign end-stage liver disease. The risk evaluation for the waiting list recipients and for post-transplant survival could provide practical indication for organ allocation. In recent years, there are two major kinds of evaluation scores. The first kind of evaluation scores is based on model for end-stage liver disease(MELD) score,including SOFT/P-SOFT score,UCLA-FRS score and BAR score. The other evaluation system is based on the concept of acute-on-chronic liver failure,including CLIF-C-ACLF score,TAM score,AARC-ACLF score and COSSH-ACLF score. The scores based on ACLF have been shown superior power in predicting waiting list survival and post-transplant prognosis than MELD. This article reviews the two kinds of evaluation scores,aiming for the better allocation policy and the better prognosis of benign end-stage liver disease.

Review of risk evaluation scores for benign end stage liver diseases recipients / 中华外科杂志

Liver transplant is an unreplaceable method for benign end-stage liver disease. The risk evaluation for the waiting list recipients and for post-transplant survival could provide practical indication for organ allocation. In recent years, there are two major kinds of evaluation scores. The first kind of evaluation scores is based on model for end-stage liver disease(MELD) score,including SOFT/P-SOFT score,UCLA-FRS score and BAR score. The other evaluation system is based on the concept of acute-on-chronic liver failure,including CLIF-C-ACLF score,TAM score,AARC-ACLF score and COSSH-ACLF score. The scores based on ACLF have been shown superior power in predicting waiting list survival and post-transplant prognosis than MELD. This article reviews the two kinds of evaluation scores,aiming for the better allocation policy and the better prognosis of benign end-stage liver disease.

Clinical characteristics of 34 COVID-19 patients admitted to intensive care unit in Hangzhou, China / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

OBJECTIVE@#This study summarizes and compares clinical and laboratory characteristics of 34 patients admitted to the intensive care unit (ICU) for complications from coronavirus disease 2019 (COVID-19) at the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China from Jan. 22 to Mar. 5, 2020.@*METHODS@#A total of 34 patients were divided into two groups, including those who required noninvasive ventilation (NIV) and invasive mechanical ventilation (IMV) with additional extracorporeal membrane oxygenation (ECMO) in 11 patients. Clinical features of COVID-19 patients were described and the parameters of clinical characteristics between the two groups were compared.@*RESULTS@#The rates of the acute cardiac and kidney complications were higher in IMV cases than those in NIV cases. Most patients had lymphocytopenia on admission, with lymphocyte levels dropping progressively on the following days, and the more severe lymphopenia developed in the IMV group. In both groups, T lymphocyte counts were below typical lower limit norms compared to B lymphocytes. On admission, both groups had higher than expected amounts of plasma interleukin-6 (IL-6), which over time declined more in NIV patients. The prothrombin time was increased and the levels of platelet, hemoglobin, blood urea nitrogen (BUN), D-dimer, lactate dehydrogenase (LDH), and IL-6 were higher in IMV cases compared with NIV cases during hospitalization.@*CONCLUSIONS@#Data showed that the rates of complications, dynamics of lymphocytopenia, and changes in levels of platelet, hemoglobin, BUN, D-dimer, LDH and IL-6, and prothrombin time in these ICU patients were significantly different between IMV and NIV cases.

Application of slender narrow pedicle flap in facial skin cancer treatment / 中华整形外科杂志

<p><b>OBJECTIVE</b>To introduce the application of slender narrow pedicle flap in repairing facial tissue defects after skin carcinoma excision, and investigate its survival mechanism.</p><p><b>METHODS</b>The slender narrow pedicle iateral maxillocevical flap was designed with its pedicle including skin fascia or only the fascia located in front of auricle or behind of it, repaired 26 cases of facial defects, including 5 temporal skin basal cell carcinoma, 6 skin squamous cell carcinoma, and 1 facial skin malignant melanoma, 8 skin basal cell carcinoma, 5 skin squamous cell carcinoma, 1 skin mucinous carcinoma. In 26 cases, 24 cases their pedicles in front of the auricle, 2 cases behind of the auricle; 4 cases their pedicles only including fascia. The size of the flaps ranged from 3.0 cm x 2.5 cm to 10.0 cm x 8.0 cm. The width and length of the pedicle ranged 1.0-1.5 cm and 2-6 cm.</p><p><b>RESULTS</b>26 cases of the slender narrow pedicle flaps all survived and the results were satisfactory except 5 cases of distal congestion, then gradual recovery.</p><p><b>CONCLUSIONS</b>This slender narrow flap don't include any major blood vessel, without dissecting the blood vessels in operating. Due to its slender pedicle, the whole flap looks like "pingpang bat", the flap rotation is easy and its coverage area is very large, without cat ears. The postoperative appearance (color, texture, cosmetic aspect) is satisfactory. This slender narrow flap is an extraordinary new flap design and is ideal for the repair of the facial tissue defect after skin carcinoma excision.</p>

Experimental study on the relationship between the ratio of length to width of slender narrow pedicle and random flap survival area in pig / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the relationship between the ratio of length to width of slender narrow pedicle and random flap survival area, and to provide a theoretical basis for the clinical application of slender narrow pedicle flaps.</p><p><b>METHODS</b>25 pigs were randomly divided into 5 groups, 5 pigs in each group. The ratio of length to width of slender narrow pedicle in 5 groups respectively was: 0:2, 1:2, 2:2, 3:2, 4:2. Every ratio' s slender narrow pedicle was carrying five different size of random flaps, which were 2 cm x 2 cm(A), 3 cm x 3 cm(B),4 cm x 4 cm(C), 5 cm x 5 cm(D), 6 cm x 6 cm(E), respectively. Flap A was control flap. In each group, flap A, B, C, D and E were created in each pigs' bilateral back. The order in both sides back is contrary. The flaps were evaluated with the general observation, fluorescence examination, blood flow ECT test, pathological expression and computerized analysis of survival area.</p><p><b>RESULTS</b>(1) The living process and pathologic process of traditional flap and slender narrow pedicle flap were consistent. It could not postpone the flap living process when the flap pedicle became long and narrow. (2) When the ratio of the length to width of the slender narrow pedicle was constant, along with the flap area increased, the flap survival area also increased, but when the flap reached a certain area , the distal flap would necrosis,the flap survival area would not reduce. (3) When the flap size remained unchanged, along with the ratio of the length to width of the slender narrow pedicle increased, the flap survival area was not affected, but when the ratio of the length to width of the slender narrow pedicle increased to a certain limit, distal flap would necrosis, the flap survival area would reduce.</p><p><b>CONCLUSIONS</b>(1) Pedicle width of random flap can be much smaller than flap width. The ratio of pedicle width to flap length is far less than traditional ratio. (2) The pedicle of random flap can be designed as slender shape, so that the whole flap looks like "pingpang bat", which makes the narrow pedicle flap rotate easily. (3) A certain ratio of the length to width of a slender narrow pedicle has a maximum flap survival area, and increasing the flap size or ratio of the length to width of a slender narrow pedicle in a certain extent will not lead to flap necrosis.</p>

Regulatory T cells inhibit proliferation of mouse lymphoma cell line EL4 in vitro / 中国实验血液学杂志

This study was aimed to investigate the effect of regulatory T (Treg) cells on the T cell lymphoma EL4 cells and its mechanism in vitro. C57BL/6 mouse Treg cells were isolated by magnetic cell sorting (MACS). The purity of Treg cells and their expression of Foxp3 were identified by flow cytometry (FCM) and PT-PCR respectively. The suppression of Treg cells on EL4 cells was detected by 3H-TdR method. At the same time, enzyme-linked immunosorbent assay (ELISA) was used to detect the secretion of cytokine TGF-β1 and IL-10. The results showed that CD4+CD25+ T cells could be successfully isolated by MACS with the purity reaching 94.52% and the expression of Foxp3 reaching 84.72%. After sorting, the expression of Foxp3 mRNA could be detected by RT-PCR. 3H-TdR assay confirmed that regulatory T cells could suppress the proliferation of EL4 cells with or without antigen presenting cells (APC) or dendritic cells (DC), APC or DC might effectively enhance the suppression. In addition, DC alone also suppressed the proliferation. TGF-β1 and IL-10 could be detected in the supernatant by ELISA. It is concluded that the Treg cells can obviously suppress the proliferation of T cell lymphoma cells in vitro, APC or DC can enhance this suppressive effect, while the DC alone also can suppress the proliferation of EL4 cells, the TGF-β1 and IL-10 cytokine pathway may be one of the mechanisms of suppression.

A method of measuring the displacement of the distal radioulnar joint on the three-dimensional CT imaging / 中华外科杂志

<p><b>OBJECTIVES</b>To establish a clinical method for measuring the displacement of the distal radioulnar joint (DRUJ) precisely irrespective of ulnar variance, and to derive normal population translation references with palmar and dorsal stress.</p><p><b>METHODS</b>Thirty-seven normal distal forearms were scanned with computed tomography using an apparatus designed by Pirela-Cruz. Each extremity was scanned in two positions: maximal ulnar palmar and dorsal stress. The digital imaging and communications in medicine (DICOM) CT images were then imported into Mimics 10.0 for three-dimensional reconstruction. On the DRUJs 3D images, choose the most prominent point of the palmar and dorsal margins of the sigmoid notch and the excavate ulna fovea as the reference points A, B and C. A perpendicular line was then drawn from the point C to a line connecting points A and B with the intersection D. Calculate the ratio of AD/AB and DB/AB. Two observers measured all the DRUJs independently and one repeated the measurements one month later to determine the interobserver and intraobserver reliability.</p><p><b>RESULTS</b>The mean ratio values of palmar (AD/AB) and dorsal (DB/AB) translation were 0.39 ± 0.07 and 0.37 ± 0.07, and the normal references (x(-) ± 2 s) were from 0.25 to 0.50 and from 0.23 to 0.50, respectively. No significant differences were observed in terms of positions, genders and dominant hands. The intraclass correlation coefficient (ICC) values for interobserver and intraobserver reliability (DB/AB, AD/AB) were 0.84, 0.80, 0.93 and 0.92, respectively.</p><p><b>CONCLUSIONS</b>This new method could accurately measure the displacement of DRUJs with acceptable reliability, even with ulna positive or negative variance. Instability of DRUJ may be indicated when AD/AB is less than 0.25 or BD/AB is less than 0.23.</p>

Island sternocleidomastoid myocutaneous flap for repairing the buccal composite tissue defect / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the application of island sternocleidomastoid myocutaneous flap in repairing the buccal composite tissue defect especially penetrated tissue defect.</p><p><b>METHODS</b>The flap pedicle included upper part OF sternocleidomastoid muscle and occipital artery. The rotation point was located at 2 cm below the mastoid. The distance between the pivot point and distal border of the defect was the length of the muscular flap. The width of the flap was slightly larger than the defect, but should not be more than 7 cm. The lower border of the flap should not exceed 2 cm below the collar bone. The flap was elevated from the starting point of the sternocleidomastoid muscle and beneath it. The pedicle only contained muscle. The flap was transferred to the defect through the tunnel between the pedicle and defect. The wounds at donor sites were closed directly or with skin graft or local flaps.</p><p><b>RESULTS</b>12 cases were treated, including 10 cases of buccal soft tissue tumors and 2 cases of buccal penetrated defects. All the musculocutaneous flaps survived with good texture, color and thickness.</p><p><b>CONCLUSIONS</b>The island sternocleidomastoid myocutaneous flap is an ideal flap for large buccal composite tissue defect with reliable blood supply. It is easily performed and very practical.</p>

Autologous regulatory T cells can suppress the proliferation of lymphoma cell line in vitro / 中国实验血液学杂志

This study was aimed to investigate the suppressive effect of regulatory T (Treg) cells on the T cell lymphoma EL4 cell line and to explore its mechanism. C57BL/6 Mouse Treg cells were isolated by MACS (magnetic cell sorting). The purity and the expression of Foxp3 were detected by flow cytometry. The suppressive effect of sorted Treg cells on EL4 cells was detected by MTT assay. The secretion of TGF-beta1 and IL-10 was examined by enzyme-linked immunosorbent assay (ELISA). The results showed that CD4(+)CD25(+) T cells could be successfully isolated by MACS with the purity reaching 91.6% and the expression level of Foxp3 was 78.9%. The ratio of viable cells was more than 95%. Regulatory T cells could suppress the proliferation of EL4 cells effectively in the presence of antigen presenting cells (APCs). And the suppressive effect was most significant at 1:1 ratio. In addition, the suppression still existed without APCs. TGF-beta1 and IL-10 could not be detected by ELISA. It is concluded that the Treg cells can suppress T lymphoma cell in vitro. The suppressive effect of Treg cells works in dose-dependent manner, but not in cytokine-dependent manner. The mechanism of this suppression may take effect through cell-cell contact.

Study on the morbidity and pathogens of patients with candidemia at the intensive care unit / 中华流行病学杂志

Objective To study the mortality and pathogens of candidemia in patients at hospital intensive care unit(ICU).Methods Clinical data of candidemia cases admitted to ICU of the First Affiliated Hospital of Zhejiang University in recent five years were analyzed retrospectively.Chi-square test and logistic regression analysis were used.Results A total of 6034 patients were discharged in the ICU over the 5-year period.and 75 were diagnosed as candidemia.The annual morbidity rates of candidemia from 2002 to 2006 were 0.67%,1.46%,1.21%,1.15% and 1.56%, respectively.36 cases died of the disease,with the mortality as 48%.The annual mortality rates from 2002 to 2006 were 50%,64%,33%,41% and 52%,respectively.In this period,78 positive blood culture samples strains from ICU were identified as Candida,among which Candida albicans, C.glabrata,C.tropicalis,C.parapsilosis and C.lusitaniae accounted for 46.2%,21.7%,17.9%,12.8% and 1.3%, respectively.Average APACHE Ⅱ scores of the patients with candidemia were 17.21±4.38(range:9-27).During the 5-year period,the annual morbidity of candidemia had increased from 0.67%to 1.56% while the ratios of candidemia due to non-albicans Candida species(NAC)increased from 50.0%to 56.5%.When analyzing the C.albicans group and NAC group with single factor and multiple conditional logistic regression method.we found that age(66±14 vs.53±16,P=0.001,OR=1.077,95% CI:1.031-1.124)and hypoproteinemia(61.8% vs.81.6%, P=0.033,OR=0.206,95%CI:0.048-0.880)both showed statistical significance.Conclusion Candidemia cases in ICU increased gradually and causing higher mortality.The number of patients with candidemia caused by NAC increased in recent 5 year.Age was proved to be a risk factor for those candidemia caused by C.albicans.Hypoproteinemia was proved to be risk factors for the candidemia caused by NAC.

Interleukin-2 improves the antitumor activity of Rituximab in patients with B-cell lymphoma in vitro / 中华血液学杂志

<p><b>OBJECTIVE</b>To establish a method to predict therapeutic effect of Rituximab and explore the feasibility and potential of IL-2 improving antitumor activity of Rituximab with upregulated antibody-dependent cellular cytotoxicity (ADCC) in patients with B-cell lymphoma.</p><p><b>METHODS</b>Eighteen B-cell lymphoma patients and 13 health volunteers entered the study. Peripheral blood mononuclear cells (PBMNC) were isolated as effector cells, and Daudi (Burkitt's lymphoma) cells as target cell, the cytotoxicity and ADCC mediated by Rituximab (10 microg/ml) of PBMNC at different E:T ratios were performed by standard 51Cr-release-assay in vitro with or without IL-2-activation. Flow cytometric analysis was performed to identify PBMNC phenotype with or without IL-2-activation.</p><p><b>RESULTS</b>A marked decrease of cytotoxicity and Rituximab mediated ADCC in the patients as compared with that in health volunteers, the results being (5.80 +/- 1.16)%, (14.32 +/- 1.50)% and (14.29 +/- 1.68)%, (24.14 +/- 1.53)% (t = 3.693, P = 0.001 and t = 3.372, P = 0.003) respectively. The decrease was correlated with the therapeutic effect of Rituximab (r = 0.781, P < 0.05). The cytotoxicity and Rituximab mediated ADCC in the patients could be partly recovered after IL-2 activation from (5.80 +/- 1.16)%, (14.32 +/- 1.50)% to (15.43 +/- 2.62)%, (35.79 +/- 2.58)% (t = 3.35, P = 0.003 and t = 7.17, P < 0.001) respectively.</p><p><b>CONCLUSIONS</b>It may be a useful method for predicting the effect of Rituximab in B-cell lymphoma patients to analyze the cytotoxicity and ADCC of their PBMNC. The impaired activity of PBMNC could be partly recovered when IL-2 was administered before the use of Rituximab.</p>

Impact of intravenous nutrition on plasma free amino acid spectrum and immune function for patients with sepsis / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the impact of intravenous nutrition on plasma free amino acid spectrum and immune function for patients with sepsis.</p><p><b>METHODS</b>Forty severe sepsis patients were divided into two groups: Group B (amino acids + glucose + fat emulsion) and Group A (glucose + fat emulsion), 20 healthy individuals were enrolled as control group. The concentration of free amino acid and immune globulin were determined after 3 days.</p><p><b>RESULTS</b>In Group A, the levels of valine, leucine, isoleucine, alanine, serine, glutamic acid, histidine, proline and glycine were decreased; while the levels of threonine, cysteine, the ratio of phenylalanine and tyrosine (Phe/Tyr) were higher than those in control group. Meanwhile, peripheral blood IgM, complement C3 and C4 were decreased. In group B, all amino acid levels were improved, but the level of alanine, serine, glutamic acid, histidine and proline still lower than those in control group. Similarly, the levels of IgM, complement C3 and C4 in group B were increased.</p><p><b>CONCLUSION</b>Intravenous nutrition can support the basal requirement of amino acid and improve the immune function of patients with sepsis.</p>

Study on monocyte HLA-DR expression in critically ill patients after surgery / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the HLA-DR expression on CD14(+) monocyte in peripheral blood of critically ill patients after surgery and observe its relationship with prognosis of patients.</p><p><b>METHODS</b>HLA-DR expression on CD14(+) monocytes in peripheral blood was measured in critically ill patients after surgery (30 cases) by using flow cytometry on the day 1, 4 and 7 after entered the ICU, and were compared with those in the healthy volunteers (28 cases). APACHE II score, sepsis-related organ failure assessment (SOFA) score, age, sex and 28 d prognosis were recorded.</p><p><b>RESULTS</b>The HLA-DR expression on CD14(+) monocyte in peripheral blood in critically ill patients after surgery were lower than that of healthy volunteers (P < 0.01). The CD14(+) monocyte HLA-DR expression on day 1 after entered ICU was not correlated with APACHE II score, SOFA and 28 d prognosis. However, compared with those with decreased HLA-DR expression, those with increased monocyte HLA-DR expression on day 7 had a better 28 d survival rate (P < 0.01).</p><p><b>CONCLUSIONS</b>In critically ill patients after surgery, the decreased HLA-DR expression on CD14(+) monocyte in peripheral blood on day 1 after entered ICU could not be regarded as a prognostic parameter, but it is significative to monitor the increased expression of HLA-DR on CD14(+) monocyte for evaluating the 28 d prognosis.</p>

Clinical characteristics and prognosis of diffuse large B-cell lymphoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the clinical characteristics of diffuse large B-cell lymphoma (DLBCL) and the factors affecting its prognosis.</p><p><b>METHODS</b>From 1994 to 2002, 138 patients with DLBCL were confirmed by morphological and immunohistochemical examination. Sex, age, clinical stage, performance status (PS), serum lactate dehydrogenase (LDH), number of extranodal lesions, treatment response, cycles of chemotherapy, B symptom, erythrocyte sedimentation rate (ESR), 5-year survival rate and median survival time (mST) were included as the analysis indeces.</p><p><b>RESULTS</b>Lymph nodes were involved in 87.7% of the patients, and extranodal lesions were found in 60.1%. Five-year survival rate was 41.3% for the entire group. Age, stage, PS, serum LDH, number of extranodal lesions, international prognostic index (IPI) and remission rates were significantly correlated with overall survival (OS) and mST (P < 0.05), However, sex, chemotherapy cycles, B symptom, ESR were not related to OS and mST (P > 0.05). Age, stage, remission rates were identified as independent factors affecting the prognosis. Combination of surgery and chemotherapy was quite impressive in the prolongation of survival of patients with extranodal lesions and gastrointestinal lymphoma compared to those by chemotherapy alone.</p><p><b>CONCLUSION</b>Age, stage, PS, serum LDH, number of extranodal lesions, IPI, chemotherapy cycles and remission rates are significant factors affecting the prognosis in DLBCL patients. Age less than 40 years or >/= 65 years, Stage III-IV, partial remission or progressive disease are demonstrated as poor prognostic factors. Combined treatment is the strategy suggested for DLBCL patients with extranodal lesions.</p>

Investigating genetic damage of workers occupationally exposed to methotrexate / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study genetic damage of workers alone occupationally exposed to methotrexate (MTX) with three end-points.</p><p><b>METHODS</b>The blood samples from 21 workers exposed to MTX and 21 controls were detected with micronucleus test, comet assay, hprt gene mutation test and TCR gene mutation test.</p><p><b>RESULTS</b>The mean micronuclei rate (MNR) and mean micronucleated cells rate (MCR) in 21 workers were 10.10 per thousand +/- 0.95 per thousand and 8.05 per thousand +/- 0.75 per thousand, respectively, which were significantly higher than those (5.48 per thousand +/- 0.82 per thousand and 4.38 per thousand +/- 0.58 per thousand) in control (P < 0.01). The mean tail length (MTL) of 21 workers and 21 controls were (1.30 +/- 0.06) microm and (0.07 +/- 0.01) microm, respectively, there was significant difference between workers and controls (P < 0.01). But the difference between workers and controls for mean tail moment (MTM) was not significant (P > 0.05). The average mutation frequency (Mf-hprt) of hprt and (Mf-TCR) of TCR in workers were 1.00 per thousand +/- 0.02 per thousand and (6.87 +/- 0.52) x 10(-4), respectively, which were significantly higher than those [0.86 per thousand +/- 0.01 per thousand and (1.67 +/- 0.14) x 10(-4)] in control (P < 0.01).</p><p><b>CONCLUSION</b>The genetic damage to some extent appeared in workers occupationally exposed to methotrexate.</p>

Change of the cell cycle after flutamide treatment in prostate cancer cells and its molecular mechanism / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To explore the effect of androgen receptor (AR) on the expression of the cell cycle-related genes, such as CDKN1A and BTG1, in prostate cancer cell line LNCaP.</p><p><b>METHODS</b>After AR antagonist flutamide treatment and confirmation of its effect by phase contrast microscope and flow cytometry, the differential expression of the cell cycle-related genes was analyzed by a cDNA microarray. The flutamide treated cells were set as the experimental group and the LNCaP cells as the control. We labeled cDNA probes of the experimental group and control group with Cy5 and Cy3 dyes, respectively, through reverse transcription. Then we hybridized the cDNA probes with cDNA microarrays, which contained 8 126 unique human cDNA sequences and the chip was scanned to get the fluorescent values of Cy5 and Cy3 on each spot. After primary analysis, reverse transcription polymerase chain reaction (RT-PCR) tests were carried out to confirm the results of the chips.</p><p><b>RESULTS</b>After AR antagonist flutamide treatment, three hundred and twenty-six genes (3.93%) expressed differentially, 97 down-regulated and 219 up-regulated. Among them, eight up-regulated genes might be cell cycle-related, namely CDC10, NRAS, BTG1, Wee1, CLK3, DKFZP564A122, CDKN1A and BTG2. The CDKN1A and BTG1 gene mRNA expression was confirmed to be higher in the experimental group by RT-PCR, while p53 mRNA expression had no significant changes.</p><p><b>CONCLUSION</b>Flutamide treatment might up-regulate CDKN1A and BTG1 expression in prostate cancer cells. The protein expressions of CDKN1A and BTG1 play an important role in inhibiting the proliferation of cancer cells. CDKN1A has a great impact on the cell cycle of prostate cancer cells and may play a role in the cancer cells in a p53-independent pathway. The prostate cancer cells might affect the cell cycle-related genes by activating AR and thus break the cell cycle control.</p>