A carbonic anhydrase-targeted NIR-II fluorescent cisplatin theranostic nanoparticle for combined therapy of pancreatic tumors.

Optically active organic nanoparticles capable of emitting strong near-infrared II (NIR-II) fluorescence and eliciting tumor hyperthermia are promising for tumor imaging and photothermal therapy (PTT). However, their applications for the treatment of pancreatic tumors via mere PTT are challenging as both the nanoparticles and light are hard to enter the deeply located pancreatic tumors. Here, we report a NIR-II light excitable, carbonic anhydrase (CA)-targeting cisplatin prodrug-decorated nanoparticle (IRNPs-SBA/PtIV) for NIR-II fluorescence imaging (FLI)-guided combination PTT and chemotherapy of pancreatic tumors. IRNPs-SBA/PtIV is designed to hold a high photothermal conversion efficiency (PCE ≈ 65.17 %) under 1064 nm laser excitation, a strong affinity toward CA (Kd = 14.40 ± 5.49 nM), and a prominent cisplatin release profile in response to glutathione (GSH) and 1064 nm laser irradiation. We show that IRNPs-SBA/PtIV can be actively delivered into pancreatic tumors where the CA is upregulated, and emits NIR-II fluorescence to visualize tumors with a high sensitivity and penetration depth under 980 nm laser excitation. Moreover, the tumor-resided IRNPs-SBA/PtIV can efficiently inhibit the CA activity and consequently, relieve the acidic and hypoxic tumor microenvironment, benefiting to intensify chemotherapy. Guided by the NIR-II FLI, IRNPs-SBA/PtIV is capable of efficiently inhibiting pancreatic tumor growth via combinational PTT and chemotherapy with 1064 nm laser excitation under a low-power density (0.5 W cm-2, 10 min). This study demonstrates promise to fabricate NIR-II excitable nanoparticles for FLI-guided precise theranostics of pancreatic tumors.

Research Progress of Chinese Medicine in the Treatment of Myocardial Ischemia-Reperfusion Injury.

Vascular recanalization is the essential procedure in which severe coronary artery stenosis is diagnosed. However, the blood flow recovery associated with this procedure may cause myocardial ischemia-reperfusion injury (MIRI), which aggravates heart failure. Unfortunately, the mechanism of MIRI has historically been poorly understood. As we now know, calcium overloading, oxidative stress, mitochondrial dysfunction, inflammatory responses, and ferroptosis take part in the process of MIRI. Modern medicine has shown through clinical studies its own limited effects in the case of MIRI, whereas Chinese traditional medicine demonstrates a strong vitality. Multiple-target effects, such as anti-inflammatory, anti-oxidant, and cardio-protection effects, are central to this vitality. In our clinic center, Yixin formula is commonly used in patients with MIRI. This formula contains Astragalus, Ligusticum Wallichii, Salvia, Rhodiola Rosea, Radix Angelicae Sinensis, Cyperus Rotundus, and Cassia Twig. Its effects include warming yang energy, activating blood circulation, and eliminating blood stasis. In our previous laboratory studies, we have proved that it can reduce MIRI and oxidative stress injury in rats suffering from ischemia myocardiopathy. It can also inhibit apoptosis and protect myocardium. In this paper, we review the research of Yixin formula and other related herbal medicines in MIRI therapy.

[Comparative study on in vivo and in vitro permeability of Huoxue Zhitong gel patch and microemulsion gel].

This study aims to establish a method for determination of paeonol(Pae), eugenol(Eug), and piperine(Pip) content in receptor liquid and research on the permeability and pharmacokinetics of Huoxue Zhitong gel patch and microemulsion gel. The Franz diffusion experiment was conducted to assess the percutaneous permeability, and the microdialysis method was employed to assess pharmacokinetics of Huoxue Zhitong gel patch and microemulsion gel. The content of Pae, Eug, and Pip in receptor liquid in vitro and in vivo was determined by HPLC and UPLC-MS. The Q_n and J_(ss) of Pae, Eug, and Pip in the gel patch were significantly higher than those in the microemulsion gel, indicating that the drug release was faster in the gel patch. The C_(max), AUC_(0-760), and MRT of Pae, Eug, and Pip in the gel patch were higher than those in the microemulsion gel, indicating that the gel patch can promote the penetration and prolong the skin residence of the drug. The results of this study provide reference for improving the dosage form of Huoxue Zhitong patch.

Anti-atopic effect of Viola yedoensis ethanol extract against 2,4-dinitrochlorobenzene-induced atopic dermatitis-like skin dysfunction.

ETHNOPHARMACOLOGICAL RELEVANCE: Viola yedoensis Makiho (VY, Violaceae) is a well-known medicinal herb in Chinese medicine, which is traditionally used to treat inflammation-related disorders, including allergic skin reactions. Although studies have uncovered its anti-inflammatory effects and corresponding bioactive constituents, the exact mechanism of action is still unclear in treating allergic skin reactions. OBJECTIVE: Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by severe pruritus, dry, edema and inflamed skin. It affects people's quality of life seriously and causes huge economic losses to society. This study proposes VY as a possible remedy for atopic dermatitis since its traditional usage and superior anti-inflammatory effects. MATERIALS AND METHODS: Atopic dermatitis-like skin lesion was induced by topical application of 2,4-dinitrochlorobenzene (DNCB) in ICR mice. After treatment with Viola yedoensis Makiho ethanol extract (VYE) or dexamethasone (positive control) for 3 weeks, skin pathological observation and the molecular biological index were performed for therapeutic evaluation, including visual inspection in the change of the stimulated skin, scar formation, pathological morphology by hematoxylin and eosin (HE) staining, the measurement of interleukin IL-1ß, IL-6 and tumor necrosis factor-alpha (TNF-α) levels in serum as well as spleen index. The expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) were analyzed by western blot. The ratio of CD4+/CD8+ T lymphocyte in the spleen was detected by flow cytometry. Meanwhile, immunohistochemistry staining for CD68 identified the number of activated macrophages in skin lesions. Additionally, a reliable ultrahigh-performance liquid chromatography coupled with a Q exactive hybrid quadrupole-orbitrap mass spectrometry (UHPLC-Q-Orbitrap-MS) method was established for the systematic identification and characterization of main components in VYE. RESULTS: VYE alleviated DNCB-stimulated AD-like lesions symptoms as evidenced by a significant decrease in hypertrophy, hyperkeratosis, and infiltration of inflammatory cells in dorsal skin. The levels of IL-1ß, IL-6, and TNF-α in serum were suppressed in mice treated with VYE as compared to the DNCB-induced model group. Also, the administration of VYE reduced the ratio of CD4+/CD8+ T lymphocyte in the spleen and the number of activated macrophages stimulated by DNCB. Besides, the expression of iNOS and COX-2 were down-regulated in the dorsal skin. CONCLUSIONS: VYE showed therapeutic effects on atopic dermatitis in DNCB-induced AD-like lesion mouse models by inhibiting the T cell-mediated allergic immune response. Our results indicated that VY could act as a potential remedy for atopic dermatitis.

[Analysis on quality value transfer of substance benchmarks of Qingwei San].

By preparing 10 batches of substance benchmarks freeze-drying powder( lyophilized powder),the methodology of the characteristic spectrum and the content of index component for substance benchmarks of Qingwei San was established. The characteristic peaks and the similarity range of the characteristic spectrum,the contents and the transfer rate range of isoferulic acid,palmatine and paeonol,and the paste-forming rate range were determined to define key quality attributes of substance benchmarks of Qingwei San. In the10 batches of substance benchmarks of Qingwei San,the similarity of characteristic spectrum was higher than 0. 90. In further comparison of the characteristic peak information,a total of 16 characteristic peaks were identified,including 5 characteristic peaks from Cimicifugae Rhizoma,5 characteristic peaks from Coptidis Rhizoma,2 characteristic peaks from Angelicae Sinensis Radix and 4 characteristic peaks from Moutan Cortex. The content of isoferulic acid was 0. 10%-0. 18%,with the average transfer rate of 49. 82%±4. 02%. The content of palmatine was 0. 17%-0. 31%,with the average transfer rate of 15. 84% ±2. 39%. The content of paeonol was 0. 41%-0. 75%,with the average transfer rate of 23. 41%±3. 23%. The paste-forming rate of the 10 batches of substance benchmarks were controlled at 27%-33%,with the transfer rate between the theoretical paste-forming rate and the actual paste-forming rate was 86. 59%±3. 39%. In this study,the quality value transfer of substance benchmarks of Qingwei San was analyzed by the combination of characteristic spectrum,the content of index component and the paste-forming rate. A scientific and stable evaluation method was preliminarily established,so as to provide the basis for subsequent development and quality control of relevant preparations of Qingwei San.

[Analysis on quality value transfer of substance benchmarks of Taohong Siwu Decoction].

By preparing 15 batches of substance benchmarks of Taohong Siwu Decoction, the methodology of the characteristic spectrums of substance benchmarks was established. The paste-forming rate range, the contents and the transfer rate range of the index components, hydroxy safflower yellow A, ferulic acid and paeoniflorin, the characteristic peaks and the similarity range of the characteristic spectrums of Taohong Siwu Decoction were determined to define key quality attributes of substance benchmarks of Taohong Siwu Decoction.In the 15 batches of substance benchmarks of Taohong Siwu Decoction, the similarity of characteristic spectrums was higher than 0.9. Furthermore, based on summarization of the characteristic peak information, there were 13 characteristic peaks in the whole decoction. Baishao had three characteristic peaks, Honghua had seven characteristic peaks, and Chuanxiong and Danggui had three characteristic peaks. The paste-forming rate of the 15 batches of substance benchmarks was controlled at 33.11%-40.62%. The content of hydroxy safflower yellow A was 0.129%-0.203%, with the average transfer rate of 16.596%±0.669%.The content of ferulic acid was 0.043%-0.055%, with the average transfer rate of 20.489%±1.772%.The content of paeoniflorin was 0.676%-0.943%, with the average transfer rate of 29.112%±3.273%.The quality value transfer of substance benchmarks of classical prescription Taohong Siwu Decoction was analyzed by the combination of characteristic spectrums, paste-forming rate and the content of index components. The established substance benchmark quality evaluation method was stable and feasible, and could provide a basis for quality control and subsequent development of relevant preparations of Taohong Siwu Decoction.

Evaluation of the wound healing potential of Sophora alopecuroides in SD rat's skin.

ETHNOPHARMACOLOGICAL RELEVANCE: Sophora alopecuroides Linn. (Leguminosae) has been largely used in traditional folk medicine in China as an anti-inflammatory agent and to treat various skin wounds, including sore furunculosis and ulcer (a common type of non-healing wound). The present study aimed to evaluate the effects of S. alopecuroides gel on skin wound healing in rats. MATERIALS AND METHODS: When the rats were anesthetized, full-thickness skin wound was performed on dorsal area by using biopsy punch with 8 mm diameter. Then, wounds received treatment with different doses of S. alopecuroides gel (1.25%, 2.5% and 5%, w/w) once a day with the gel base used as vehicle control and rb-FGF as positive control. Every five animals were sacrificed after 7, 12 days after surgery for histopathology and relevant biochemical indexes analysis. Besides, after RAW 264.7 cells exposure to LPS (1 µg/ml) with or without total extract (25 and 50 µg/ml) for 24 h, the culture supernatant was used for detection of IL-1ß and TNF-α levels using ELISA kits and the protein lysate for western blot analysis. RESULTS: A remarkable wound closure was observed after administration with 5% S. alopecuroides gel with the wound area of 30% and 8.5% as compared to 42% and 19% in the control group on day 7 and 12, respectively. Histological and immunostaining analysis for the wound tissues also revealed that S. alopecuroides promoted the growth of granulation tissue, collagen deposition, cell proliferation and angiogenesis. Meanwhile, it was able to ameliorate inflammatory response and promote the production of TGF-ß. In addition, we also demonstrated that S. alopecuroides inhibited the release of inflammatory mediators and expression of iNOS as well as up-regulated the expression of Arg-1 in LPS-triggered RAW 264.7 cells. CONCLUSIONS: The present study confirmed that S. alopecuroides had a great potential for accelerated wound healing by regulating the over expression of inflammatory response for the first time and provided theoretical basis for the traditional use. It can be used as candidate drug for the treantment of chronic non-healing wounds.

Analysis of 70 patients with hydrocephalus due to cobalamin C deficiency.

OBJECTIVE: To analyze the clinical characteristics of patients with hydrocephalus secondary to cobalamin C (cblC) deficiency and to discuss the optimal strategies for assessing and treating such patients by performing clinical and laboratory studies in 70 patients. METHODS: A total of 1,211 patients were clinically diagnosed with methylmalonic acidemia (MMA) from 1998 to 2019. Among them, cblC deficiency was confirmed in 70 patients with hydrocephalus by brain imaging and biochemical and genetic analysis. RESULTS: Of the 70 patients, 67 (95.7%) had early-onset MMA and homocystinuria. The patients typically had high blood propionylcarnitine and total homocysteine, low methionine, and methylmalonic aciduria. Signs of intracranial hypertension were relatively rare. We measured ventricular dilatation early in the disease by cranial ultrasound and MRI and/or CT. Eighteen different MMACHC mutations, including 4 novel mutations (c.427C>T, c.568insT, c.599G>A, and c.615C>A), were identified biallelically in all 70 patients. c.609G>A was the most frequent mutation, followed by c.658_660del, c.217C>T, and c.567dupT. Three cases were diagnosed by postmortem study. Metabolic therapy, including cobalamin injections supplemented with oral l-carnitine and betaine, was administered in the remaining 67 cases. A ventriculoperitoneal shunt was performed in 36 cases. During the follow-up, psychomotor development, nystagmus, impaired vision, and sunset eyes improved gradually. CONCLUSION: Hydrocephalus is a severe condition with several different causes. In this study, ventriculomegaly was found in 70 patients with cblC deficiency. Early diagnosis, etiologic treatment, and prompt surgical intervention are crucial to improve the prognosis of patients.

Double-layer substrate of shale ceramsite and active alumina tidal flow constructed wetland enhanced nitrogen removal from decentralized domestic sewage.

This study explored the application of shale ceramsite (SC) overlaid onto active alumina (AA) to function as a double-layer substrate in tidal flow constructed wetland (TFCW, SC-AA-TFCW) for decentralized domestic sewage treatment. This was compared to AA or SC substrate alone (AA-TFCW or SC-TFCW) for nitrogen removal, variation of dissolved oxygen and porosity in TFCWs, as well as structure of bacterial communities at varied hydraulic load (HL) of 0.204-2.448 m3/m2 d with time ratio of the wet and dry phase of 3:1. The results demonstrate that SC-AA-TFCW removed 86% NH4+-N and 79% total nitrogen at HL of 0.612 m3/m2 d, which was better than AA-TFCW(76%) or SC-TFCW(49%). The higher nitrogen removal performance in SC-AA-TFCW was mainly attributed to enhanced oxygen transportation due to non-uniform flow field and irregular gap distribution in layered structure, as well as less pore blockage during long-term operation. Denitrifying bacteria including Dechloromonas, Acidovorax, Chryseobacterium and Thermomonas species took up 32% of the microbiome in SC-AA-TFCW, which was higher than in AA-TFCW (17%) and SC-TFCW (7.7%). This study highlighted the importance of layered structures and determined an optimal HL of TFCW to achieve an efficient and stable nitrogen removal for domestic sewage treatment.

A novel mutation associated with Type III Bartter syndrome: A report of five cases.

The clinical, biochemical and mutation spectra of Chinese patients with Type III Bartter syndrome (type III BS), a rare autosomal recessive disorder, were investigated. A total of five unrelated Chinese patients aged 8 months to 24 years were diagnosed with type III BS via analysis of biochemical markers, including chloride, potassium and calcium, and genetic sequencing. The levels of insulin­like growth factor­1 (IGF­1) were evaluated via ELISA and a mutation study of cultured amniocytes was conducted for prenatal diagnosis. The child patients were admitted for polydipsia, polyuria, myasthenia and developmental delay, whereas the adult patients were hospitalized for limb numbness, polydipsia and polyuria. Nine variants in the chloride voltage­gated channel Kb (CLCNKB) gene were detected, including eight sequence variants and one whole CLCNKB gene deletion. One sequence variant (c.1967T>C) was novel, whereas the remaining variants (c.595G>T, c.908A>C, c.1004T>C, c.1312C>T, c.1334_1335delCT and c.1718C>A) and the whole gene deletion had been previously reported. The whole gene deletion was frequently observed in patients with early­onset type III BS in the present study. Two patients showed IGF­1 deficiency with normal growth hormone level. All patients were treated with potassium supplementation and indometacin. The mother of one patient underwent amniocentesis during her second pregnancy; the fetus was not affected by type III BS based on screening for sequence variants, and normal development and blood electrolyte analysis following birth confirmed the diagnosis. In conclusion, five cases of type III BS in patients from mainland China were reported. Large deletions were frequently detected, particularly in early­onset patients; isolated IGF­1 deficiency was found, one novel sequence variant was identified. Prenatal diagnosis was successfully established using genetic analysis of cultured amniocytes, and may facilitate the prevention of congenital defect of type III BS in the next pregnancy.

A Chinese pedigree with Brown-Vialetto-Van Laere syndrome due to two novel mutations of SLC52A2 gene: clinical course and response to riboflavin.

BACKGROUND: Brown-Vialetto-Van Laere Syndrome (BVVLS), a rare neurological disorder characterized by motor, sensory, and cranial neuronopathies, is mainly associated with defective riboflavin transporters encoded by SLC52A2 and SLC52A3 genes. Clinical outcomes have been shown to be improved significantly by high-dose riboflavin supplementation. The aim of this study was to identify genetic causes and further evaluate the clinical course and response to riboflavin in a Chinese pedigree with BVVLS. CASE PRESENTATION: We report the novel compound heterozygous variants c.1328G>A p.(Cys443Tyr) and c.1022_1023insC p. (Leu341Profs*103) of SLC52A2 gene in a female proband who presented in our out-patient clinic at the age of one-year-old with progressive mental and motor regression, breath holding, and brain stem dysfunction including facial weakness, hearing loss, dysphagia. Following high-dose riboflavin supplementation, the respiratory insufficiency and mental, motor, and bulbar function improved. However, sensorineural hearing loss was not improved. The missense variant site was highly conserved. Both variants were not found in the population database gnomAD. The two variants were inherited from her mother and father, respectively. Both variants were predicted to be deleterious by Polyphen2, Mutation taster, and SIFT and were classified as likely pathogenic according to the ACMG guideline. CONCLUSIONS: Two novel pathogenic variations of SLC52A2 gene were firstly found from a Chinese pedigree with BVVLS. Clinical outcomes could be improved by early diagnosis and riboflavin supplementation.

Enhanced simultaneous organics and nutrients removal in tidal flow constructed wetland using activated alumina as substrate treating domestic wastewater.

A tidal flow constructed wetland (TFCW), a commonly applied system to clean wastewater, contains a substrate to assist pollutants removal, while the choice of substrate affects the formation of bacterial biofilms. Herein, activated alumina-TFCW (A-TFCW) with hydraulic load of 1.35 m3/(m2·d) parallel with shale ceramisite (S-TFCW) was investigated for treating domestic wastewater, aiming to enhance simultaneous long-term removal of organics, nitrogen and phosphorus. A-TFCW achieved significantly higher COD, NH4+-N, TN and TP removal efficiency than S-TFCW, with the removal efficiency of 85.9% COD, 85.4% NH4+-N, 72.8% TN and 96.4% TP respectively. Denitrifying bacteria dominated in both formed biofilms, with higher relative abundance of nitrifying bacteria and denitrifying bacteria in A-TFCW. These results demonstrated that AA substrate was more suitable to be applied in enhancing the removal performance in TFCW for the treatment of domestic wastewater.

Patients with cobalamin G or J defect missed by the current newborn screening program: diagnosis and novel mutations.

Cobalamin G (cblG) and cobalamin J (cblJ) defects are rare disorders of cbl metabolism caused by MTR and ABCD4 mutations, respectively. Patients with atypical biochemical features can be missed by current newborn screening using tandem mass spectrometry (MS/MS), in which total homocysteine (tHCY) in dried blood spots (DBS) is not a primary biomarker. Two Chinese patients suspected of cbl defect but missed by newborn screening were studied. Using comprehensive metabolic analyses including MS/MS assay for tHCY in DBS, slightly low methionine in Patient 1, methymalonic aciduria in Patient 2, and homocysteinemia in both patients were detected, and DBS tHCY of two patients were obviously elevated (59.22 µmol/L, 17.75 µmol/L) compared to 140 healthy controls (2.5th-97.5th percentile, 1.05-8.22 µmol/L). Utilizing whole-exome sequencing, we found two novel MTR variants c.871C>T (p.Pro291Ser) and c.1771C>T (p.Arg591*) in Patient 1, and a ABCD4 homozygous variant c.423C>G (p.Asn141Lys) in Patient 2. Our study identified the first cblG patient and cblJ patient in mainland China, and highlighted comprehensive metabolic analyses and genetic tests in patients suspected of cbl defects. It also indicated that supplementary MS/MS assay for tHCY in DBS may be practical for early diagnosis of homocysteinemia, without repeated blood sampling.

[Effects of acetyl-L-carnitine on the recovery of hindlimb movement afterspinal cord injury in rats and its mechamism].

OBJECTIVE: To observe the effects of different doses of acetyl-L-carnitine (ALC) on hindlimb motor function and spinal cord tissue structure in rats with spinal cord injury. The study will provide theoretical and experimental evidences for acetyl-L-carnitine's clinical treatment. METHODS: Fifty-five SD rats aged 8-10 weeks were randomly divided into high, medium and low-dose drug intervention (SCI + ALC) group, injury group (SCI) and sham group for behavioral evaluation, MAD and SOD detection, as well as HPLC detection and HE staining. BBB scores and Rivlin experiments were performed to evaluate hindlimb motor function in each group. The morphology and structure of spinal cord tissue was detected by HE staining. Another 9 rats were randomly divided into Sham group, SCI group and ALC group for TUMEL detection of apoptosis. RESULTS: The BBB scores of the high, medium, and low dose SCI+ALC groups were significantly higher than those in the SCI group. The medium and high-dose ALC groups had significant differences (P＜0.01), and the hindlimb motor function was significantly improved in rats. The maximum tilt angle of the Rivlin experiment was observed. The SCI+ALC group had a significantly increased angle compared with the SCI group (P＜0.05), the medium and high-dose ALC group had a significant difference (P＜0.01). Compared with the SCI group, the tissue structure of ALC high-dose group was improved significantly, the number of inflammatory cells and red blood cells was decreased, and the nucleolus of the nerve cells was unclear. The SOD activity of the SCI+ALC group was significantly higher than that of the SCI group, while the MDA content was significantly decreased(P＜0.05), the middle and high dose ALC groups were significantly different (P＜0.01). HPLC chromatogram showed that the SCI+ALC fresh serum sample and the ALC standard solution had the same absorption spectrum at 260 nm, while the Sham group and SCI group serum samples did not show spectral values there, which indicated that the same substance as the standard existed in the sample of SCI+ALC group. TUNEL staining showed that the apoptosis signal was occasionally seen in the sham group, and the apoptosis signal was significantly decreased in the ALC high-dose group compared with the SCI group(P＜0.05). CONCLUSION: ALC can promote the recovery of hindlimb motor function in rats with spinal cord injury, inhibit oxidative stress and apoptosis, and repair the damaged spinal cord tissue.

Strategic Combination of Isocratic and Gradient Elution for Simultaneous Separation of Polar Compounds in Traditional Chinese Medicines by HPLC.

A simple high-performance liquid chromatography (HPLC) method for the simultaneous separation of the highly polar and weakly polar components of traditional Chinese medicines was developed via a strategic combination of isocratic and gradient elution methods. Liu-Shen-Wan and Liu-Wei-Di-Huang-Wan were used as representative examples of traditional Chinese medicines. This is the first time that 6 components of varying degrees of polarity in Liu-Shen-Wan had been successfully resolved in a single chromatographic run using an ultraviolet-visible detector with a fixed wavelength of 296 nm. In contrast to conventional gradient separation methods, this novel method offered a viable route for separation of the highly and weakly polar fractions simultaneously, thus greatly reducing the time and cost of analysis. This method therefore provides a more efficient way to determine the polar components present in traditional Chinese medicines. It would find potential application in drug screening, drug authentication, and product quality control.

[Acute encephalopathy induced by vaccination in an infant with methylmalonic aciduria cblA].

OBJECTIVE: We report the first case of acute encephalopathy induced by vaccination in an infant with methylmalonic aciduria cblA in China. METHOD: The clinical presentation, blood acylcarnitines analysis, urine organic acids analysis and gene studies of the patient were summarized. RESULT: The proband, a boy, was admitted at the age of 15 months because of recurrent vomiting, acidosis and development delay for 8 months. The previously healthy boy presented vomiting and coma just one hour after hepatitis B vaccination at the age of seven months. Moderate dehydration, electrolyte disturbance and metabolic acidosis had been found. Although his acute metabolic crisis had been corrected soon after intravenous transfusion, psychomotor retardation and recurrent vomiting had been observed. When he was 15 months old, vomiting and lethargy occurred again 3 hours after DTaP vaccination. He was weakened as the illness became worse and got coma with dyspnea 7 days later. He was hospitalized with the suspected diagnosis of viral encephalitis. Blood acylcarnitines analysis, urine organic acids analysis and gene study had been performed for the etiologic investigation.His blood propionylcarnitine (16.3 µmol/L vs. normal range 1.0-5.0 µmol/L) and propionylcarnitine/free carnitine ratio (0.27 vs. normal range 0.03 to 0.25) increased. Markedly elevated urinary methylmalonic acid (388.21 mmol/mol creatinine vs. normal range 0.2 to 3.6 mmol/mol creatinine) and normal plasma total homocysteine supported the diagnosis of isolated methylmalonic aciduria. Two mutations, c.650 T>A (p.L217X) and c.742 C>T (p.Q248X), were identified in his MMAA gene, confirmed the diagnosis of cblA. Each parent carried one of the two mutations. Progressive clinical and biochemical improvement has been observed after hydroxylcobalamin injection, protein-restricted diet with the supplements of special formula and L-carnitine. He is currently 2 years and 7 months old with normal development and general condition. CONCLUSION: A boy with cblA was firstly detected after the acute encephalopathy induced by vaccination in China. It is important to pay more attention to the patients with metabolic crisis or organ damage after vaccination. Metabolic studies are keys to the diagnosis of potential diseases and improve the outcome.

Very long-chain acyl-coenzyme A dehydrogenase deficiency in Chinese patients: eight case reports, including one case of prenatal diagnosis.

OBJECTIVE: Very long-chain acyl-coenzyme A dehydrogenase deficiency (VLCADD) is a rare mitochondrial fatty acid ß-oxidation disorder. We aimed to explore the clinical, biochemical, and genetic findings, treatments and outcomes in eight Chinese VLCADD patients. METHODS: Eight patients from six unrelated Chinese families with symptomatic VLCADD were diagnosed in the past 4 years. The clinical features and ACADVL gene mutations were analyzed. RESULTS: One patient underwent newborn screening and has been treated timely, she hardly had any symptoms. The remaining seven patients were found because of edema, diarrhea, coma, liver damage and psychomotor retardation. Seven patients had fatty liver. Five had myopathy. All patients had elevated blood tetradecanoylcarnitine. Nine heterozygous mutations of the ACADVL gene were found. Three (c.1102C > T, c.1795G > A and IVS10, +6T > A) were novel. Seven patients completely recovered after treatment. One patient died before diagnosis due to cardiomyopathy. His mother underwent amniocentesis for prenatal diagnosis. The fetus had the same gene mutation of the proband and markedly elevated tetradecanoylcarnitine in amniotic fluid. The boy has been treated after birth and he is healthy now. CONCLUSIONS: Dietary treatment usually leads to good outcomes to VLCADD patients. Amniocytes ACADVL mutations and amniotic fluid tetradecanoylcarnitine analysis are useful for the prenatal diagnosis.

The first Chinese case report of hereditary folate malabsorption with a novel mutation on SLC46A1.

BACKGROUND: Hereditary folate malabsorption is a rare, autosomal recessive disorder of proton-coupled folate transporter deficiency resulting in folate deficiency. Left untreated, the condition can cause severe brain damage and megaloblastic anemia, leading to progressive psychomotor retardation, seizures and other neurological problems. Early diagnosis and treatment are crucial. No case has been documented yet in Mainland China until now. METHODS: A Chinese girl affected by hereditary folate malabsorption was studied. The girl presented with recurrent megaloblastic anemia from the age of 7 months. Paroxysmal limbs trembling and seizures were presented from the age of three years. Intracranial calcification was noted by CT. At her age of 5 years, mental regression, lower-extremity weakness and sleeping problems were observed. Her plasma folate decreased to 4.49 nmol/L (normal control>6.8nmol/L). Plasma total homocysteine elevated to 28.11 µmol/L (normal control<15 µmol/L). Folate and 5-methylterahydrofolate in cerebrospinal fluid were significantly decreased to undetectable level. RESULTS: On SLC46A1 gene, a novel mutation, c.1A>T (M1L), and a reported mutation c.194-195 insG (p.Cys66LeufsX99) were identified, supported the diagnosis of hereditary folate malabsorption. Each parent carries one of two mutations. Folinic calcium supplement resulted in rapid clinical improvement. She is currently 6 years old with normal development and routine blood features. CONCLUSION: Hereditary folate malabsorption is one of the few easily-treatable inherited metabolic diseases. Measurements of folate and 5-methyltetrahydrofolate in cerebrospinal fluid are keys for the diagnosis of the patients.

Involvement of substance p/neurokinin-1 receptor in the analgesic and anticancer activities of minimally toxic fraction from the traditional Chinese medicine Liu-Shen-Wan in vitro.

Liu-Shen-Wan (LSW), an ancient preparation used to treat localized infection with pain, was recently reported to possess anticancer activity. The mechanism responsible for LSW's analgesic and anticancer activity is unclear. In the present study, we obtained a LSW supernatant (LSWS) fraction from ultrasound-assisted ethanol extraction (yield 15.9%) which proved to be safer than LSW in terms of hepatotoxicity. The LSWS (1 and 10 µg/mL) exhibited a potent inhibitory effect on the bradykinin-evoked rapid release of substance P from dorsal root ganglion (DRG) cells. At concentrations of 0.1 µg/mL and higher, the LSWS resulted in a concentration-related growth inhibitory effect on HepG2, a representative cancer cell lines. The LSWS significantly down-regulated the neurokinin-1 (NK-1) receptor expression in both HepG2 and bradykinin-treated DRG cells. In addition to the NK-1 receptor-dependent growth inhibition in HepG2 cells (0.1-100 µg/mL), the LSWS induced mitochondria-mediated apoptosis at a higher concentration (1-100 µg/mL). In conclusion, we recently isolated a safer LSW fraction which maintained its analgesic and anticancer activity, and found that the substance P/NK-1 receptor system was partly responsible for these effects. Our findings will be useful for developing more effective and less toxic LSW preparations.