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IntroductionSelenium is an essential micronutrient the human body requires, which is closely linked to health. Rice, a primary staple food globally, is a major source of human selenium intake. To develop selenium-enriched rice varieties, it is imperative to understand the mechanisms behind selenium’s absorption and transport within rice, alongside identifying the key genes involved in selenium uptake, transport, and transformation within the plant.MethodsThis study conducted transcriptome sequencing on four types of rice materials (two with low-selenium and two with high-selenium contents) across roots, stems, leaves, and panicles to analyze the gene expression differences.Results and discussionDifferential gene expression was observed in the various tissues, identifying 5,815, 6,169, 7,609, and 10,223 distinct genes in roots, stems, leaves, and panicles, respectively. To delve into these differentially expressed genes and identify the hub genes linked to selenium contents, weighted gene co-expression network analysis (WGCNA) was performed. Ultimately, 10, 8, 7, and 6 hub genes in the roots, stems, leaves, and panicles, respectively, were identified. The identification of these hub genes substantially aids in advancing our understanding of the molecular mechanisms involved in selenium absorption and transport during the growth of rice.

Tibetan tea changes during microorganism fermentation. Research on microorganisms in Tibetan tea has focused on their identification, while studies on the influence of specific microorganisms on the components and health functions of Tibetan tea are lacking. Bacillus licheniformis was inoculated into Tibetan tea for intensive fermentation, and the components of B. licheniformis-fermented tea (BLT) were detected by liquid chromatography with tandem mass spectrometry (UHPLC-TOF-MS), and then the effects of BLT on intestinal probiotic functions were investigated by experiments on mice. The results revealed the metabolites of BLT include polyphenols, alkaloids, terpenoids, amino acids, and lipids. Intensified fermentation also improved the antioxidant capacity in vivo and the protective effect on the intestinal barrier of Tibetan tea. In addition, the enhanced fermentation of Tibetan tea exerted intestinal probiotic effects by modulating the relative abundance of short-chain fatty acid-producing bacteria in the intestinal flora. Therefore, intensive fermentation with B. licheniformis can improve the health benefits of Tibetan tea.

To investigate the dynamic changes and potential correlations between microbial diversity and volatile organic compounds (VOCs) during Chinese medium-temperature Daqu (MTD) manufacturing at different key stages, in this study, high-throughput sequencing (HTS) and gas chromatography–ion mobility spectrometry (GC–IMS) were employed to analyze the microbial diversity and VOCs of MTD, respectively. The results showed that Weissella, Staphylococcus, Thermoactinomyces, Kroppenstedtia, and Lactobacillus were the dominant bacterial genera, while Aspergillus, Alternaria, Thermoascus, Thermomyces, Wickerhamomyces, and Saccharomyces were the dominant fungal genera. A total of 61 VOCs were detected by GC–IMS, among which, 13 differential VOCs (VIP > 1) were identified, that could be used as potential biomarkers to judge the fermentation stage of MTD. Kroppenstedtia and Saccharopolyspora were positively correlated with 3-methyl-2-butenal and 2,2,4,6,6-pentamethylheptane-D, respectively, and both of these were positively correlated with butanal-D. Acetobacter, Streptomyces, and lactic acid bacteria (LAB) including Leuconostoc, Pediococcus, Weissella, and Lactobacillus were negatively correlated with their associated VOCs, while fungi were generally positively correlated with VOCs. Wickerhamomyces, Saccharomyces, and Candida were positively correlated with butan-2-one-M. This study provides a theoretical basis for explaining the mechanisms of MTD flavor formation and screening functional microorganisms to improve the quality of MTD.

In this study, we investigated the effect of solid-state fermentation of Pleurotus eryngii on the composition and antioxidant activity of Tartary buckwheat (TB). Firstly, the solid-state fermentation of P. eryngii mycelium with buckwheat was carried out, and the fermentation process was explored. The results of the extraction process and method selection experiments showed that the percolation extraction method was superior to the other two methods. The results of extraction rate, active components and antioxidant activity measurements before and after fermentation of TB extract showed that the extraction rate increased about 1.7 times after fermentation. Total flavonoids, rutin and triterpene contents were increased after fermentation compared to control. Meanwhile, LC-MS results showed an increase in the content of the most important substances in the fermented TB extract and the incorporation of new components, such as oleanolic acid, ursolic acid, amino acids, and D-chiral inositol. The fermented TB extract showed stronger antioxidant activity, while the protein and amino acid contents increased by 1.93-fold and 1.94-fold, respectively. This research was the first to use P. eryngii to ferment TB and prepared a lyophilized powder that could be used directly using vacuum freeze-drying technology. Not only the use of solid-state fermentation technology advantages of edible fungi to achieve value-added buckwheat, but also to broaden the scope of TB applications. This study will provide ideas and directions for the development and application of edible mushroom fermentation technology and TB.

Objective: To study the effect of β-nicotinamide mononucleotide (NMN) on the growth of Caenorhabditis elegans (C. elegans) and its mechanism. Methods: Nematodes were randomly divided into blank control group and experimental groups with different concentrations of NMN (0.1, 0.5, 1, 5, 10 mg/mL). The impact of NMN on the growth of C. elegans was evaluated by measuring the body length changes of C. elegans in 13 days after adulthood. In addition, transcriptomics was used to explore the mechanism of NMN (1 mg/mL) in promoting nematode growth. Moreover, quantitative real-time polymerase chain reaction (qRT-PCR) was performed to verify the sequencing results. Results: Compared with the blank control group, the average body length of C. elegans was significantly increased after feeding 0.1, 0.5, 1, 5 and 10 mg/mL NMN at 4 to 13 days (P

Abstract Identification of the core functional microorganisms in food fermentations is necessary to understand the ecological and functional processes for making those foods. Wheat qu, which provides liquefaction and saccharifying power, and affects the flavor quality, is a key ingredient in ancient alcoholic huangjiu fermentation, while core microbiota of them still remains indistinct. In this study, metagenomics, metabolomics, microbial isolation and co-fermentation were used to investigate huangjiu. Although Aspergillus is usually regarded as core microorganism in wheat qu to initiate huangjiu fermentations, our metagenomic analysis showed that bacteria Saccharopolyspora are predominant in wheat qu and responsible for breakdown of starch and cellulose. Metabolic network and correlation analysis showed that Saccharopolyspora rectivirgula, Saccharopolyspora erythraea, and Saccharopolyspora hirsuta made the greatest contributions to huangjiu’s metabolites, consisting of alcohols (phenylethanol, isoamylol and isobutanol), esters, amino acids (Pro, Arg, Glu and Ala) and organic acids (lactate, tartrate, acetate and citrate). S. hirsuta J2 isolated from wheat qu had the highest amylase, glucoamylase and protease activities. Co-fermentations of S. hirsuta J2 with S. cerevisiae HJ resulted in a higher fermentation rate and alcohol content, and huangjiu flavors were more similar to that of traditional huangjiu compared to co-fermentations of Aspergillus or Lactiplantibacillus with S. cerevisiae HJ. Genome of S. hirsuta J2 contained genes encoding biogenic amine degradation enzymes. By S. hirsuta J2 inoculation, biogenic amine content was reduced by 45%, 43% and 62% in huangjiu, sausage and soy sauce, respectively. These findings show the utility of Saccharopolyspora as a key functional organism in fermented food products.

The diversity of alcohol beverage microorganisms is of great significance for improving the brewing process and the quality of alcohol beverage products. During the process of making alcoholic beverages, a group of microorganisms, represented by yeast and lactic acid bacteria, conducts fermentation. These microorganisms have complex synergistic or competitive relationships, and the participation of different microorganisms has a major impact on the fermentation process and the flavor and aroma of the product. Strain selection is one of the key steps. Utilizing scientific breeding technology, the relationship between strains can be managed, the composition of the alcoholic beverage microbial community can be improved, and the quality and flavor of the alcoholic beverage products can be increased. Currently, research on the microbial diversity of alcohol beverages has received extensive attention. However, the selection technology for dominant bacteria in alcohol beverages has not yet been systematically summarized. To breed better-quality alcohol beverage strains and improve the quality and characteristics of wine, this paper introduces the microbial diversity characteristics of the world’s three major brewing alcohols: beer, wine, and yellow wine, as well as the breeding technologies of related strains. The application of culture selection technology in the study of microbial diversity of brewed wine was reviewed and analyzed. The strain selection technology and alcohol beverage process should be combined to explore the potential application of a diverse array of alcohol beverage strains, thereby boosting the quality and flavor of the alcohol beverage and driving the sustainable development of the alcoholic beverage industry.

In order to reduce the interference of seasonal, geographical and other factors and explore the effect of different peak-temperature control strategies on the fungal community of Daqu, this study mainly applied the high-throughput sequencing technology to analyze and compare the fungal community in medium-temperature and high-temperature Daqus collected from the same core production region of high-quality Baijiu. The correlation between the fungal community in Daqu and the main physicochemical indices was also evaluated by combining redundancy analysis. The results showed that the overall population of fungi (characterized by the copy numbers of ITS1 region of rDNA) in high-temperature Daqu was smaller than that in medium-temperature Daqu, but the richness, diversity, and evenness of overall fungal community in the former were higher than that in the latter. Thermoascus, Pichia, Aspergillus and Rhizopus accounted for a higher proportion in medium-temperature Daqu, with Thermoascus aurantiacus as the absolute advantage. Thermomyces, Rasamsonia, Monascus, and Byssochlamys accounted for a higher proportion in high-temperature Daqu, with Thermomyces lanuginosus as the absolute advantage. Based on the random forest prediction model, the first ten key ASVs that could best explain the differences between the two Daqu communities belonged respectively to three major groups including Thermoascus crustaceus (for 5 ASVs), Thermomyces spp. (for 4 ASVs), and Thermoascus aurantiacus (for 1 ASV). The results of redundancy analysis showed that Aspergillus, Rasamsonia and Hyphopichia in two types of Daqu had a positive correlation with saccharifying power, Pichia had a strong positive correlation with acidity, and Thermoascus had a strong positive correlation with moisture. This study further clarified the diversity of fungal community and biomarkers in Daqus with different peak-temperatures, explored the correlation between the fungal community in Daqu and the main physicochemical indices, and provided a reference for the optimization of Daqu making process and the screening of functional strains.

To satisfy the demands of rapid determination of amylose, amylopectin, protein, fat, and tannin contents in brewing sorghums, in this paper, 17 spectral data preprocessing methods and 4 wavelength band selection algorithms were used to establish the near infrared spectral analysis models for these indexes. The results showed that the best spectral preprocessing methods for each index were 1st der (1st)+multiplicative scatter correction (MSC)+Z-score standardization (ZS), vector normalization (VN)+mean centering (MC), standard normal variate transformation (SNV)+ZS, MSC, SNV+ZS, respectively. The best wavelength band selection algorithm for predicting amylose, amylopectin, protein, and tannin contents was monte-carlo uninformative variable elimination, and that of fat was competitive adaptive reweighted sampling. The R2 in the optimal models for these 5 indexes of whole grain sorghums were 0.9560, 0.8765, 0.9069, 0.8658, 0.8841, and the RMSECV values were 1.3222, 2.3477, 0.3549, 0.2164, 0.1077, respectively. The validation results of external independent samples showed that the models had a high prediction accuracy. The NIR analysis model established in this study could provide a technical reference for the rapid detection of sorghums in the brewing industry.

Distiller’s grains resourcing is a crucial problem for the sustainable development of the Chinese Baijiu industry. In order to improve the protein utilization of distiller’s grains. In this study, distiller’s grains pretreated by wet crushing combined with amylase/cellulase were further hydrolyzed with neutral protease combined with papain at 50 °C for 3 h. The hydrolysates were decolorized by 1% coconut shell activated carbon for 15 min, and separated and purified by macroporous resin and Sephadex gel to yield peptide fractions (F1, F2, F3 and F4). The F3 fraction with the highest radical scavenging activity was analyzed by nano-LC-MS/MS and peptidomics to identify peptides. Three peptides (especially newly discovered AAHVLAAAFL) displayed strong alcohol solubility and antioxidant activity and were stably bound to key regulatory pathway receptors (Keap1-Nrf2) and myeloperoxidase (MPO), further elucidating molecular mechanism of anti-oxidation. These findings provided knowledge for potential functional peptide candidates and facilitates the high-value commercial development of distiller’s grains.

Abstract Background Osteonecrosis of the femoral head (ONFH) is still a challenge for orthopedists worldwide and can lead to disability if patients are not treated effectively. Danyu Gukang Pill (DGP), a traditional Chinese medicine (TCM) formulation, is recognized to be effective against ONFH. Nevertheless, its molecular mechanisms remain to be clarified. Methods The active ingredients of DGP were collected from the online databases according to oral bioavailability (OB) and drug-likeness (DL). The potential targets of DGP were retrieved from the TCMSP database, while the potential targets of ONFH were obtained from the GeneCards and NCBI databases. The functions and signaling pathways of the common targets of DGP and ONFH were enriched by GO and KEGG analyses. Subsequently, molecular docking and in vitro cell experiments were performed to further validate our findings. Results In total, 244 active ingredients of DGP and their corresponding 317 targets were obtained, and 40 ONFH-related targets were predicted. Afterwards, 19 common targets of DGP and ONFH were obtained and used as potential targets for the treatment of ONFH. Finally, combined with network pharmacology analysis, molecular docking and in vitro cell experiments, our study first demonstrated that the treatment effect of DGP on ONFH might be closely related to the two targets, HIF1A (HIF-1α) and VEGFA, and the HIF-1 signaling pathway. Conclusions This study is the first to investigate the molecular mechanisms of DGP in the treatment of ONFH based on network pharmacology. The results showed that DGP might up-regulate the expression of HIF-1α and VEGFA by participating in the HIF-1 signaling pathway, thus playing an anti-ONFH role.

Abstract It is of great significance for quality control to realize the discrimination for baijiu from different brands and origins. Strong-aroma-type baijiu (SAB), one of the most important Chinese aroma-type baijiu, exhibits the largest variety and market share. In this study, we proposed colorimetric sensor arrays based on gold nanoparticles (AuNPs) modified with different amino acids (AAs) to recognize the organic acids, and further distinguish different SABs. Three representative AAs, namely methionine (Met), tryptophan (Trp), and histidine (His), were selected to modify the AuNPs surface. The investigation of the effect of the main ingredients of SAB on AA@AuNPs aggregation confirmed that this aggregation mainly resulted from organic acids. Moreover, this aggregation was successfully used for differentiating 11 organic acids. Different pH conditions can not only cause changes of the content of organic acids in baijiu, but also disrupt the balance among flavor substances of baijiu to some extent. Consequently, the AA@AuNPs arrays under two pH conditions have been successfully applied to distinguish 14 kinds of SABs from different brands and origins. The proposed colorimetric sensor method is simple, rapid, and visualized and provides a potential application prospect for the quality control of baijiu and other alcoholic beverages.

Luxiang-flavor Baijiu is the mainstream of Baijiu production and consumption in China, and the microbial composition has a great influence on the flavor and quality of Baijiu. In this study, we combined multi-omics sequencing technology to explore the microbial composition, dynamics and metabolite changes of Luxiang-flavor Jiupei during long fermentation periods. The results showed that based on the interaction between environmental constraints and microorganisms, Jiupei microorganisms formed different ecological niches and functional differentiation, which led to the formation of Jiupei stable core microorganisms. The bacteria were mainly Lactobacillus and Acetobacter, and the fungi were mainly Kazachstani and Issatchenkia. Most bacteria were negatively correlated with temperature, alcohol and acidity, and for the fungi, starch content, reducing sugar content and temperature had the most significant effects on community succession. Macroproteomic analysis revealed that Lactobacillus jinshani had the highest relative content; microbial composition, growth changes and functions were more similar in the pre-fermentation period (0–18 days); microorganisms stabilized in the late fermentation period (24–220 days). The metabolome analysis revealed that the metabolites of the Jiupei changed rapidly from 18 to 32 days of fermentation, with a significant increase in the relative content of amino acids, peptides and analogs and a significant decrease in the relative content of sugars; the metabolites of the Jiupei changed slowly from 32 to 220 days of fermentation, with a stabilization of the content of amino acids, peptides and analogs. This work provides insights into the microbial succession and microbial drivers during the long-term fermentation of Jiupei, which have potential implications for optimizing production and improving the flavor of Baijiu.

Liquid-liquid extraction (LLE) is the most commonly utilized technique for the extraction of odor-active esters (OAEs) in strong-aroma types of Baijiu (SAB). However, since the contents of different OAEs in SAB vary widely, it is still a puzzle to ensure that all OAEs to be thoroughly extracted by LLE without the problem of saturated adsorption. Herein, a novel approach of magnetic solid phase extraction (MSPE), based on the magnetic graphene oxide nanocomposite modified with polyacrylamide (GO/PAM/Fe3O4), was employed for the efficient extraction of six OAEs from SAB. Compared with LLE, GO/PAM/Fe3O4 exhibited highly selective recognition properties and larger adsorption capacities for OAEs (ranging from 13.68 to 39.06 mg/g), resulting in better extraction performances for OAEs. Coupled with GC–MS, six OAEs in real SAB were successfully determined, with recoveries ranged from 70.1 ∼ 90.0% and LODs at 0.08 ∼ 1.35 µg/L. Overall, the MSPE-GC/MS is a promising alternative for accurate determination of OAEs in SAB.

Chinese Baijiu is one of the famous distilled liquor series with unique flavors in the world. Under the open environment, Chinese Baijiu was produced by two solid-state fermentation processes: jiuqu making and baijiu making. Chinese Baijiu can be divided into different types according to the production area, production process, starter type, and product flavor. Chinese Baijiu contains rich flavor components, such as esters and organic acids. The formation of these flavor substances is inseparable from the metabolism and interaction of different microorganisms, and thus, microorganisms play a leading role in the fermentation process of Chinese Baijiu. Bacteria, yeasts, and molds are the microorganisms involved in the brewing process of Chinese Baijiu, and they originate from various sources, such as the production environment, production workers, and jiuqu. This article reviews the typical flavor substances of different types of Chinese Baijiu, the types of microorganisms involved in the brewing process, and their functions. Methods that use microbial technology to enhance the flavor of baijiu, and for detecting flavor substances in baijiu were also introduced. This review systematically summarizes the role and application of Chinese Baijiu flavor components and microorganisms in baijiu brewing and provides data support for understanding Chinese Baijiu and further improving its quality.

Although a large number of trials in the SCI field have been conducted, few proven gains have been realized for patients. In the present study, we determined the efficacy of a novel combination treatment involving surgical intervention and long-term weight-bearing walking training in spinal cord injury (SCI) subjects clinically diagnosed as complete or American Spinal Injury Association Impairment Scale (AIS) Class A (AIS-A). A total of 320 clinically complete SCI subjects (271 male and 49 female), aged 16–60 years, received early (≤ 7 days, n = 201) or delayed (8–30 days, n = 119) surgical interventions to reduce intraspinal or intramedullary pressure. Fifteen days post-surgery, all subjects received a weight-bearing walking training with the “Kunming Locomotion Training Program (KLTP)” for a duration of 6 months. The neurological deficit and recovery were assessed using the AIS scale and a 10-point Kunming Locomotor Scale (KLS). We found that surgical intervention significantly improved AIS scores measured at 15 days post-surgery as compared to the pre-surgery baseline scores. Significant improvement of AIS scores was detected at 3 and 6 months and the KLS further showed significant improvements between all pair-wise comparisons of time points of 15 days, 3 or 6 months indicating continued improvement in walking scores during the 6-month period. In conclusion, combining surgical intervention within 1 month post-injury and weight-bearing locomotor training promoted continued and statistically significant neurological recoveries in subjects with clinically complete SCI, which generally shows little clinical recovery within the first year after injury and most are permanently disabled. This study was approved by the Science and Research Committee of Kunming General Hospital of PLA and Kunming Tongren Hospital, China and registered at ClinicalTrials.gov (Identifier: NCT04034108) on July 26, 2019.

Rubia cordifolia L. (Rubiaceae), one of the traditional anti-rheumatic herbal medicines in China, has been used to treat rheumatoid arthritis (RA) since ancient times. Purpurin, an active compound of Rubia cordifolia L., has been identified in previous studies and exerts antibacterial, antigenotoxic, anticancer, and antioxidant effects. However, the efficacy and the underlying mechanism of purpurin to alleviate RA are unclear. In this study, the effect of purpurin on inflammation was investigated using macrophage RAW264.7 inflammatory cells, induced by lipopolysaccharide (LPS), and adjuvant-induced arthritis (AIA) rat was established to explore the effect of purpurin on joint damage and immune disorders; the network pharmacology and molecular docking were integrated to dig out the prospective target. Purpurin showed significantly anti-inflammatory effect by reducing the content of IL-6, TNF-α, and IL-1β and increasing IL-10. Besides, purpurin obviously improved joint injury and hypotoxicity in the liver and spleen and regulated the level of FOXP3 and CD4+/CD8+. Furthermore, purpurin reduced the MMP3 content of AIA rats. Network pharmacology and molecular docking also suggested that MMP3 may be the key target of purpurin against RA. The results of this study strongly indicated that purpurin has a potential effect on anti-RA.

Sauce-aroma Baijiu (SAB) is one of the most famous Baijius in China; SAB has more than 500 aroma compounds in it. However, the key aroma compound in SAB flavor remains unclear. Volatiles play an important role in SAB aroma and are highly correlated to SAB quality. In the present study, 63 volatile compounds were quantified among 66 SAB samples using gas chromatography with flame ionization detector (GC-FID). The authors analyzed odor contributions and volatile compound correlations in two quality groups of SAB samples. Moreover, an odor activity value (OAV) ratio-based random forest classifier was used to explain the volatile compound relationship differentiations between the two quality groups. Our results proved higher quality SABs had richer aromas and indicated a set of fruity-like ethyl valerate, green- and malt-like isobutyraldehyde and malt-like 3-methylbutyraldehyde and sweet-like furfural, had closer co-abundance correlations in higher quality SABs. These results indicated that the aroma and contributions of volatile compounds in SABs should be analyzed not only with compound odor activity values, but also the correlations between different aroma compounds.

Surface-enhanced Raman spectroscopy (SERS) is an emerging technology that combines Raman spectroscopy and nanotechnology with great potential. This technology can accurately characterize molecular adsorption behavior and molecular structure. Moreover, it can provide rapid and sensitive detection of molecules and trace substances. In practical application, SERS has the advantages of portability, no need for sample pretreatment, rapid analysis, high sensitivity, and ‘fingerprint’ recognition. Thus, it has great potential in food safety detection. Alcoholic beverages have a long history of production in the world. Currently, a variety of popular products have been developed. With the continuous development of the alcoholic beverage industry, simple, on-site, and sensitive detection methods are necessary. In this paper, the basic principle, development history, and research progress of SERS are summarized. In view of the chemical composition, the beneficial and toxic components of alcoholic beverages and the practical application of SERS in alcoholic beverage analysis are reviewed. The feasibility and future development of SERS are also summarized and prospected. This review provides data and reference for the future development of SERS technology and its application in food analysis.

Fusel alcohols (FAs) are a type of flavor compound found in rice wine. An overly high FA content not only leads to spicy, bitter, and astringent taste but also has side effects. Therefore, screening for yeast that produce low FA contents has attracted much attention. Thirty-two yeast strains were isolated from fermenting material during Luzhou-flavor liquor production in this study. Strain YB-12 was selected as a suitable candidate for rice wine production. The strain was identified as a member of the genus Meyerozyma based on phylogenetic analysis using 26S rDNA gene sequences. The ability of strain YB-12 to produce ethanol was similar to that of Saccharomyces cerevisiae NRRL Y-567, while isobutanol and isoamyl alcohol production was only 53.96% and 50.23%, respectively, of that of NRRL Y-567. The FA yield of rice wine produced with strain YB-12 was reduced to 51.85% in a 20 L fermenter. These results demonstrate that strain YB-12 presents promising characteristics for use in the production of rice wine with a potentially low content of FAs.

Umbilical cord blood-derived mononuclear cell (UCB-MNC) transplants improve recovery in animal spinal cord injury (SCI) models. We transplanted UCB-MNCs into 28 patients with chronic complete SCI in Hong Kong (HK) and Kunming (KM). Stemcyte Inc. donated UCB-MNCs isolated from human leukocyte antigen (HLA ≥4:6)-matched UCB units. In HK, four patients received four 4-μl injections (1.6 million cells) into dorsal entry zones above and below the injury site, and another four received 8-μl injections (3.2 million cells). The eight patients were an average of 13 years after C5-T10 SCI. Magnetic resonance diffusion tensor imaging of five patients showed white matter gaps at the injury site before treatment. Two patients had fiber bundles growing across the injury site by 12 months, and the rest had narrower white matter gaps. Motor, walking index of SCI (WISCI), and spinal cord independence measure (SCIM) scores did not change. In KM, five groups of four patients received four 4-μl (1.6 million cells), 8-μl (3.2 million cells), 16-μl injections (6.4 million cells), 6.4 million cells plus 30 mg/kg methylprednisolone (MP), or 6.4 million cells plus MP and a 6-week course of oral lithium carbonate (750 mg/day). KM patients averaged 7 years after C3-T11 SCI and received 3–6 months of intensive locomotor training. Before surgery, only two patients walked 10 m with assistance and did not need assistance for bladder or bowel management before surgery. The rest could not walk or do their bladder and bowel management without assistance. At about a year (41–87 weeks), WISCI and SCIM scores improved: 15/20 patients walked 10 m ( p = 0.001) and 12/20 did not need assistance for bladder management ( p = 0.001) or bowel management ( p = 0.002). Five patients converted from complete to incomplete (two sensory, three motor; p = 0.038) SCI. We conclude that UCB-MNC transplants and locomotor training improved WISCI and SCIM scores. We propose further clinical trials.