[Comparison of the characteristics of NK cells after two different methods of expansion and observation of the clinical efficacy in patients who relapsed post allogeneic hematopoietic stem cell transplantation].

Objective: To explore the differences in the biological effects of different expansion systems on natural killer (NK) cells, as well as the safety and preliminary clinical efficacy in the treatment of patients with recurrence after allogeneic hematopoietic stem cell transplantation (allo-HSCT) . Methods: Peripheral blood cells from healthy donors were stimulated with either CD3 combined with CD52 or K562 feeder cells loaded with IL-21/4-1BB to induce NK cell expansion. Changes in the NK cell phenotype, cytokine secretion, and cytotoxicity before and after expansion were detected. We also evaluated the safety and clinical efficacy of two different expansion strategies for patients received NK infusion. Results: Compared with the CD3/CD52 monoclonal antibody amplification system, the feeder cell expansion group had a higher purity of NK cells and higher expression ratios of NK cell surface activation receptors such as DNAM-1 and NKp30, while inhibitory receptor CTLA-4 expression was low and NKG2D/CD25/CD69/ Trail/PD-1/TIM-3/TIGIT had no statistically significant differences between the groups. Further functional results showed that the expression level of KI67 in NK cells after expansion in the two groups increased significantly, especially in the feeder cell expansion group. Simultaneously, the perforin and granzyme B levels of NK cells in the feeder cell expansion group were significantly higher than in the CD3/CD52 expansion group. A retrospective analysis of eight patients who received monoclonal antibody-expanded NK cell reinfusion and nine patients with trophoblast cell-expanded NK cell reinfusion was done. The disease characteristics of the two groups were comparable, NK cell reinfusion was safe, and there were no obvious adverse reactions. Clinical prognostic results showed that in the CD3/CD52 monoclonal antibody amplification group, the MRD conversion rate was 50% (2/4) , and the feeder cell expansion group was 50% (3/6) . After 5 years of follow-up from allo-HSCT, three patients in the monoclonal antibody expansion group had long-term survival without leukemia, and the remaining five patients had died; two patients died in the feeder cell expansion group, and the other six patients had long-term survival. Six cases had GVHD before NK cell reinfusion, and GVHD did not aggravate or even relieved after NK cell reinfusion. Conclusions: Preliminary results show that the biological characteristics of NK cells with diverse expansion strategies are significantly different, which may affect the clinical prognosis of patients with recurrence or persistent minimal residual disease after HSCT. The two groups of patients treated with NK cells from different expansion strategies had no obvious adverse reactions after NK cell infusion, but efficacy still needs to be further confirmed.

[Prevalence of deep venous thrombosis in chronic refractory wounds and its influencing factors].

Objective: To investigate the incidence and risk factors of lower extremities deep venous thrombosis (DVT) in patients with chronic refractory wounds (chronic wounds). Methods: A retrospective analysis of 722 cases of chronic wounds admitted to the First People's Hospital of Zhengzhou from January 1, 2015 to December 31, 2018 was carried out, and clinical data such as gender, age, wound involves the lower extremities, wound type, duration of wound, D-dimer level and complicated diabetes, hypertension, coronary heart disease, cerebral infarction, pneumonia and sepsis/septic shock were collected. The patients were divided into DVT group and non-DVT group according to the color ultrasound test. Multivariate binary logistic regression analysis was used to screen the independent risk factors of DVT in patients with chronic wounds. Results: There were 55 (7.6%) patients in DVT group, of which 44(80.0%) were diagnosed as chronic DVT by conventional lower extremities venous color ultrasound screening at admission, and the remaining 11(20.0%) were newly diagnosed during hospitalization. There were no significant differences in gender, wound involves the lower extremities, duration of wound, complicated diabetes and hypertension between DVT group and non-DVT group (all P>0.05); and the differences were statistically significant in age, wound type, D-dimer level, complicated coronary heart disease, cerebral infarction, pneumonia and sepsis/septic shock (all P<0.05). Multivariate binary logistic regression analysis showed age [OR=2.260 (95%CI: 1.584-3.224)], D-dimer level [OR=14.201 (95%CI: 6.101-33.055)], complicated coronary heart disease [OR=2.731 (95%CI: 1.210-6.162)] and sepsis/septic shock [OR=3.170 (95%CI: 1.115-9.016)] were independent risk factors for DVT (all P<0.05). Conclusion: Chronic wounds patients have a higher incidence of DVT, most of which are chronic DVT, and age, D-dimer level, complicated coronary heart disease and sepsis/septic shock are independent risk factors for DVT in chronic wounds.

Raloxifene retards cartilage degradation and improves subchondral bone micro-architecture in ovariectomized rats with patella baja-induced - patellofemoral joint osteoarthritis.

OBJECTIVE: Abnormal remodeling of subchondral bone (SB) induced by estrogen deficiency has been shown to be involved in osteoarthritis (OA). Raloxifene (RAL) is commonly used to treat postmenopausal osteoporosis (OP). However, little is known about its effects on OA combined with estrogen deficiency. This study was performed to evaluate the efficacy of RAL on patella baja-induced patellofemoral joint OA (PFJOA) in an ovariectomized rat model. DESIGN: Patellar ligament shortening (PLS) and ovariectomy (OVX) were performed simultaneously in 3-month-old female Sprague-Dawley rats, which were treated with RAL (10 mg/kg/day) or vehicle at 72 h postoperatively for 10 weeks. PFJOA was assessed by immunohistochemistry (IHC), real-time polymerase chain reaction (PCR), tartrate-resistant acid phosphatase (TRAP) staining, enzyme-linked immunosorbent assay (ELISA), micro-computed tomography (µCT), histomorphology and behavioral analyses. RESULTS: X-ray examinations showed that patella baja was successfully established by PLS. Histomorphological analysis revealed that PFJOA was significantly exacerbated by OVX and markedly alleviated by RAL. Moreover, RAL improved cartilage metabolism by decreasing MMP-13, ADAMTS-4, and caspase-3 and increasing Col-II and aggrecan at both the protein and mRNA levels. Furthermore, RAL markedly improved bone mass and SB microarchitecture and reduced osteoclast numbers and the serum osteocalcin and CTX-I levels. Although RAL showed a trend toward reducing pain sensitivity based on mechanical allodynia testing, this result was not statistically significant. CONCLUSION: These findings demonstrate that RAL treatment retards PFJOA progression in an ovariectomized rat model, suggesting that it may be a potential candidate for amelioration of the progression of PFJOA accompanied by postmenopausal OP.

Effect of developmental stage of embryos at freezing on live birth outcomes after frozen embryo transfer.

　　Purpose Our objective was to evaluate the birth outcomes of FET from different developmental stage of embryo at freezing (Day 3, Day 5 and Day 6). Methods All vitrified-warmed day 3 (D3) and day 5/6 (D5, D6) embryo transfers during the time period from January 2013 until December 2015 were included in the study. The Birthweight, Low birthweight (LBW), sexual distinction, gestation and for singleton births and twins conceived by FET were compared retrospectively between three cryopreservation strategies utilizing either D3, D5 or D6 embryo freezing. All embryos were vitrified by Dimethylsulfoxide (DMSO), Ethylene glycol (EG) and Sucrose used as Cryoprotectants and using Cryoleaf as the receptacles. Results A total of 993 infants from 771 women. The length of embryos storage was 3-38 months, with a median of 8 months. For singleton, birthweight from D5 and D6 blastocysts transfers were significantly heavier than for D3 cleavage-stage embryos transfers (P =0.0065 P =0.0006). For twins, birthweight from D6 blastocysts transfers were significantly heavier than for D5 blastocysts transfers (P =0.0044), and children born after D3 were at a signigicantly increased risk of being born a LBW. Conclusions Birthweights from FET are influenced by developmental stage of embryos at freezing, which were lower for cleavage-stage embryos transfer than blastocysts transfer after FET in singletons. The birth rate of LBW infants was higher in the twins. The study was small so there may be other factors than cryopreservation which affected outcomes. A higher sample size or a multi-centre prospective randomized design could be used in future studies to corroborate the current findings.

Direct high-precision measurement of the effective optical path length of multi-pass cell with optical frequency domain reflectometer.

Multi-pass cells (MPCs) are commonly used in trace-gas detection and weak spectrum measurement. It is essential to accomplish a high-precision measurement of MPCs' effective optical path length (EOPL). A direct high-precision measuring method of MPCs' EOPL with optical frequency domain reflectometer (OFDR) was reported and demonstrated in this paper. Several important parameters of a MPC, such as EOPL and base length, were derived with high-precision by identifying the complicated signal of OFDR. The MPC's EOPL was also verified with the prevailing absorbance method. The results showed that the MPC's EOPL measured by each of these two methods is highly consistent. However, the relative uncertainty with the OFDR dramatically decreased 2 orders of magnitude (about 0.0085%) than that with the absorbance method. It demonstrated that the OFDR method with fewer measurement links is more conducive to a direct measurement. The performances of beam spread and stray light in the White-cell were also evaluated with the method.

Initiation of active immunization against testosterone during early puberty alters negative feedback regulation of the hypothalamic-pituitary-testicular axis in rabbits.

To investigate the effects of antitestosterone immunization, initiated during early puberty, on hypothalamic-pituitary-testicular feedback in rabbits, 16 early pubertal male rabbits were randomly allocated into 2 groups (n = 8), control or immunized against testosterone-3(O-carboxymethyl)oxime-BSA in Freund adjuvant at 4 mo of age (with a booster immunization 4 wk later). Blood samples (for antibody titers and hormone concentrations) were collected at 2- or 4-wk intervals after immunization. Compared with controls, antitestosterone immunization triggered: a substantial and sustained antibody response (P < 0.01); increases in serum concentrations of luteinizing hormone (LH) and testosterone and testis weight and volume (P < 0.05); hyperplasia of testicular interstitial tissue with clustered and hypertrophic Leydig cells; and greater (P < 0.05) enzyme protein and messenger RNA (mRNA) expression levels for testicular cholesterol side-chain cleavage cytochrome P-450, 17α-hydroxylase cytochrome P-450, and 3ß-dydroxysteroid dehydrogenase. Furthermore, immunoneutralization of testosterone upregulated mRNA expressions for genes in sex steroid negative feedback loops, including androgen receptor (AR), estrogen receptor alpha (ER-α), kisspeptin encoded gene (kiss-1) and kisspeptin receptor (G-coupled receptor 54) and gonadotropin-releasing hormone (GnRH) in the hypothalamic arcuate nucleus, GnRH receptor and LH-ß in pituitary, and AR, inhibin-α and ßA subunits in testes (P < 0.05). However, immunization did not affect mRNA expressions for follicle-stimulating hormone ß, AR, and ER-α in pituitary, or ER-α in testes. We concluded that antitestosterone immunization in male rabbits, initiated during early puberty, increased GnRH mRNA expression, and in turn LH synthesis by reducing testicular feedback signaling. Reduction of direct steroidal effects on the testis may also have increased testosterone secretion. Consequently, there was an accelerated testicular development during puberty and enhanced testicular function after puberty, which likely conferred prolonged reproductive advantages.

The association of new inflammatory markers with type 2 diabetes mellitus and macrovascular complications: a preliminary study.

AIM: Type 2 diabetes mellitus (T2DM) is not simply a disease of hyperglycemia, but also is an inflammatory disorder. This study aimed to observe the expression of inflammation-related factors in elderly T2DM patients with or without macrovascular disease (MVD). PATIENTS AND METHODS: A total of 64 T2DM patients participated in this study, including 31 patients with MVD (group A) and 33 patients without MVD (group B); and 30 healthy volunteers were recruited as normal control (group C). The levels of serum irisin, retinol-binding protein 4 (RBP4) and adiponectin expression were all detected and compared between groups. RESULTS: The demographic and clinical characteristics were comparable between T2DM patients and healthy volunteers. For patients in group A, the serum levels of irisin, RBP4 and adiponectin were 12.05 ± 2.12 pg/mL, 2.13 ± 0.83 µg/mL and 45.65±20.13 ng/mL, respectively. While the corresponding parameters were 26.11 ± 4.09 pg/ml, 1.54 ± 0.54 µg/ml and 57.93 ± 23.47 ng/mL for patients in group B; and were 40.25 ± 2.73 pg/mL, 0.98 ± 0.36 µg/mL and 60.03 ± 20.26 ng/mL for healthy volunteers in group C, respectively. As compared to healthy volunteers, the levels of irisin, RBP4 and adiponectin were all significantly changed in T2DM patients; and the difference in irisin, RBP4 and adiponectin between T2DM patients with and without MVD were all significant (p = 0.000, p = 0.001, and p = 0.029, respectively). Multivariate regression analysis showed that irisin and RBP4 are both independent predictors for MVD in T2DM patients. CONCLUSIONS: Inflammatory disorder is significantly in T2DM patients with MVD, and serum irisin and RBP4 would be reasonable new markers of MVD.

The Spirodela polyrhiza genome reveals insights into its neotenous reduction fast growth and aquatic lifestyle.

The subfamily of the Lemnoideae belongs to a different order than other monocotyledonous species that have been sequenced and comprises aquatic plants that grow rapidly on the water surface. Here we select Spirodela polyrhiza for whole-genome sequencing. We show that Spirodela has a genome with no signs of recent retrotranspositions but signatures of two ancient whole-genome duplications, possibly 95 million years ago (mya), older than those in Arabidopsis and rice. Its genome has only 19,623 predicted protein-coding genes, which is 28% less than the dicotyledonous Arabidopsis thaliana and 50% less than monocotyledonous rice. We propose that at least in part, the neotenous reduction of these aquatic plants is based on readjusted copy numbers of promoters and repressors of the juvenile-to-adult transition. The Spirodela genome, along with its unique biology and physiology, will stimulate new insights into environmental adaptation, ecology, evolution and plant development, and will be instrumental for future bioenergy applications.

Degradation of nitenpyram pesticide in aqueous solution by low-temperature plasma.

In order to study the new technique of plasma wastewater treatment, the degradation behaviour ofnitenpyram (NTP) pesticide was investigated in a low-temperature plasma formed during a dielectric barrier discharge process. The reactor was a radial flow sedimentation tank centred around the water inlet. We studied the effect of pesticide concentration and input power of the dielectric barrier discharge, together with the effect of external factors on the degradation of nitenpyram pesticide wastewater such as conductivity and the use of various of catalysts, and the reaction products were analyzed by high-performance liquid chromatography mass spectrometry (HPLC-MS). The results showed that NTP could be effectively removed from aqueous solution by low-temperature plasma. Increasing the input power could improve the efficiency of degradation, conforming to a first-order kinetic model. Use of a suitable catalyst clearly improved the degradation process, as also did low conductivity. The pH of NTP was reduced with discharge time.

CLP induces apoptosis in human leukemia K562 cells through Ca(2+) regulating extracellular-related protein kinase ERK activation.

The cyclic lipopeptide (CLP) has been known to inhibit proliferation and induce apoptosis in cancer cells. However, the molecular mechanisms involved in CLP-induced apoptosis are still uncharacterized in human leukemic K562 cells. The current study investigated the molecular mechanism of action of CLP, purified from Bacillus natto T-2. CLP-induced a sustained increase in concentration of intracellular Ca(2+). This increase in [Ca(2+)]i was associated with CLP-induced cell apoptosis and ERK phosphorylation. CLP-induced cell apoptosis was reversed by PD98059 (an inhibitor of ERK), but not by SB203580 (an inhibitor of p38) and SP200125 (an inhibitor of JNK), suggesting that the action of CLP on K562 cells was via ERK, but not via p38 and JNK. On the other hand, pretreatment with Bapta-AM, a well-known calcium chelator, partially blocked CLP-induced apoptosis, indicating that the elevation of [Ca(2+)]i may play an important role in the apoptosis. Collectively, in K562 cells, CLP-induced an increase in [Ca(2+)]i which evoked ERK phosphorylation. This ERK phosphorylation subsequently activated Bax, cytochrome c and caspase-3 leading to apoptosis.

[Determination of puerarin in xinmaitong oral liquid by HPLC].

OBJECTIVE: To develop a method to determine the content of puerarin in Xinmaitong oral liquid. METHODS: HPLC method was used. The separation was performed on Suntek kromasil C18 column with methanol-water containing 0.5% acetic acid (25:75) as a mobile phase and the wavelength of UV detector was 250 nm. RESULTS: The linearity of puerarin was good and average recovery was 97.8%. CONCLUSION: The method is simple, reliable and sensitive. It also shows good resolation. It can be used in quality control of Xinmaitong oral liquid.

Race-specific HIV-1 disease-modifying effects associated with CCR5 haplotypes.

Genetic variation in CC chemokine receptor 5 (CCR5), the major HIV-1 coreceptor, has been shown to influence HIV-1 transmission and disease progression. However, it is generally assumed that the same CCR5 genotype (or haplotype) has similar phenotypic effects in different populations. To test this assumption, we used an evolutionary-based classification of CCR5 haplotypes to determine their associated HIV-1 disease-modifying effects in a large well-characterized racially mixed cohort of HIV-1-seropositive individuals. We demonstrate that the spectrum of CCR5 haplotypes associated with disease acceleration or retardation differs between African Americans and Caucasians. Also, we show that there is a strong interactive effect between CCR5 haplotypes with different evolutionary histories. The striking population-specific phenotypic effects associated with CCR5 haplotypes emphasize the importance of understanding the evolutionary context in which disease susceptibility genes are expressed.

Selective total vascular occlusion for major hepatic resection.

Selective total vascular occlusion for the resection of large tumors on the right lobe of the liver or central hepatic tumors was described. The occlusion of the portal triad, infrahepatic vena cava and the total hepatic circulation was selectively used in combination to control bleeding in the process of hepatectomy. Within a year, major resection was successfully performed with this technique on 10 patients (extended right lobectomy 4 patients, right lobectomy 4, central segmentectomy 2). The mean duration of total vascular exclusion was 19.35 +/- 19.32 minutes and mean blood transfusion requirement was 7 +/- 4.5 units during surgery.

[Determination of neuropeptide Y concentration in the cerebrospinal fluid in patients with cerebrovascular diseases and its significance].

Concentration of neuropeptide Y (NPY) in the cerebrospinal fluid (CSF) of patients with cerebrovascular diseases was measured by using radioimmunoassay. The results showed that NPY concentration in CSF in patients with hemorrhagic cerebrovascular diseases (HCVD) was 4148 +/- 397.2 pg/ml, being significantly higher than the control level of 1083.7 +/- 245.8 pg/ml. While the NPY concentration in CSF of patients with ischemic cerebrovascular diseases (ICVD) was 2214 +/- 289.2 pg/ml, being not significantly different from the level in the control group. The effect of NPY in vasospasm after HCVD was preliminarily discussed.

[Modified normothermic avascular hepatic resection].

The conventional Heaney's avascular hepatic resection was modified based on an experimental study on normothermic total hepatic vascular exclusion in dog. The modification was cancellation of simultaneous exclusion of the abdominal aorta and proper prolongation of total hepatic vascular exclusion time. The method was successfully used in 3 patients, 2 with primary hepatocellular carcinoma involving the second porta hepatis and 1 with a huge hepatic cavernous hemangioma in segments VII and VIII. The clinical application of this method is presented and discussed. The modified Heaney's technique, being simpler and safer than the conventional one, may increase the resection rate for central hepatic tumors (segments I, IV, V and VIII according to Couinaud's classification).

Experimental research on utilization of bioluminescent technique to substitute the current viability count for BCG vaccine.

The possibility of using the bioluminescent (BL) technique to substitute the traditional viability count of colony forming units (CFU) of BCG vaccine was investigated. The results showed there is a significant dose-dependent correlation between the concentration of standard adenosine triphosphate (ATP) and ATP BL value. The ultrasonic-chloroform method designed by us yielded the best results. The correlation coefficient values (r) of BL of the liquid and lyophilized vaccine were found to be 0.8 155 and 0.8 484 respectively (P less than 0.05). The coefficient of variation (CV) between the BL value of different lots of ATP obtained was 3.2-4.4%, much lower than that of CFU (CV = 10.4-11.2%). The presence of bacterial clumps within the vaccine had great influence on the bacterial ATP value and on the CFU viability count. 2.8 fg of ATP was found in each CFU formed in the vaccine with clumps, while it was only 0.94 fg in the vaccine without clumps, indicating the superiority of the BL method. The BL method has shown a high sensitivity, good reproducibility and simplicity in handling with quick results and high accuracy. Therefore, we consider that the BL method can be used to substitute the CFU method.