OBJECTIVETo emphasize early differential diagnosis from patients with hyperphenylalaninemia (HPA) and to evaluate the treatment and long-term outcome of patients with tetrahydrobiopterin synthase (BH4) deficiency in Northern Chinese population.

METHODSFrom 1992 to 2005, a total of 618 patients with HPA were diagnosed and/or cared for in our outpatient clinic. Urinary pterin analysis, detection of dihydropteridine reductase (DHPR) activity in blood, and then BH4 loading tests were carried out to differentiate BH4 deficiency in these patients from classical phenylketonuria. BH4 deficient patients were treated with BH4, levodopa and 5-hydroxytryptophane (5-HTP) immediately while the diagnosis was done to disease. Patientso blood phenylalanine levels, psychomotor and intelligence development were followed up.

RESULTSA total of 38 cases were diagnosed as BH4 deficiency, all of them were revealed as 6-pyruvoyl-tetrahydropterin synthase (PTPS) deficiency from the extremely decreased urine biopterin, normal DHPR activities and drop down of blood phenylalanine level to normal range within 4 to 8 hours after BH4 loading. The most common manifestations were progressively psychomotor and mental retardation to patients even after taking early dietary treatment. The patients were diagnosed and treated with drugs at the ages of 2.1 months to 13 years. With 4 patients died of pneumonia, 7 patients refused to treatment, only 27 patients were under treatment and followed up. The average full scale development or intelligence quotient (DQ/IQ) of patients who were treated within and after 6 months were 86+/- 10 or 66+/- 7 respectively. Development was not even in different aspects. A significant negative correlation was observed between the level of the DQ and the age of treatment commenced (r was -0.714, P< 0.01). Eleven patients experienced the extrapyramidal movement disorders, 3 of them combined with epilepsy. The extrapyramidal disorders were controlled by administration of levodopa.

CONCLUSIONThe differential diagnosis for BH4 deficiency should be carried out in all patients with HPA. PTPS deficiency is the most common form of BH4 deficiency in Northern Chinese population. The long-term outcome of these patients benefits from diagnosis and treatment with BH4, levodopa and 5-HTP as early as possible.

5-Hydroxytryptophan ; therapeutic use ; Asian Continental Ancestry Group ; genetics ; Biopterin ; analogs & derivatives ; deficiency ; therapeutic use ; Child, Preschool ; China ; Dihydropteridine Reductase ; blood ; Humans ; Infant ; Levodopa ; therapeutic use ; Phenylalanine ; blood ; Phenylketonurias ; drug therapy ; genetics ; metabolism ; Phosphorus-Oxygen Lyases ; deficiency ; genetics

Objective By using resting-state fMRI (r-fMRI), to observe the effect of electroacupuncture at Quchi (LI 11) and Shangjuxu (ST 37) in changing brain activity pattern in functional bowel disorders (FBD), and to provide imaging evidence for seeking the central mechanism of Quchi (LI 11) and Shangjuxu (ST 37) in treating FBD.Method Twenty patients with functional constipation were taken as the constipation group, and 20 patients with functional diarrhea were taken as the diarrhea group. The constipation group was intervened by electroacupuncture at Shangjuxu (ST 37), while the diarrhea group was intervened by electroacupuncture at Quchi (LI 11). On the next day, the two groups both received electroacupuncture at the corresponding non-meridian point. The r-fMRI data of the two groups were collected before and after the interventions. With the functional connectivity taken as the observation index and right insular lobe as the seed point, MTLAB, DPARSF and REST were adopted to obtain the brain images functionally connected with the seed point, to further analyze the difference in brain functional connectivity before and after electroacupuncture.Result After electroacupuncture at Shangjuxu (ST 37), functional constipation patients presented positive activation in left cerebellar declive, left lingual gyrus, left middle occipital gyrus, right occipital lobe, right supramarginal gyrus, right gyrus cingula and left precuneus; the right subgyrus, right insular lobe and left insular lobe were negatively evoked. After electroacupuncture at Quchi (LI 11), patients with functional diarrhea showed significant differences in left cerebellar culmen and left posterior cingulate.Conclusion Electroacupuncture at Quchi (LI 11) and Shangjuxu (ST 37) can induce changes in brain functional connectivity related to visceral hypersensitivity in FBD patients, which is possibly associated with the effect of acupuncture in improving FBD.

ObjectiveTo investigate the effect of Yiqi Jiedu prescription-containing serum on the proliferation of medullary thymic epithelial cells （mTEC） and regulatory T （Treg） cells in myasthenia gravis （MG） patients with thymus hyperplasia. MethodAccording to serological methods，35 SD rats were adaptively fed for one week and randomized into the low-，medium-， and high-dose Yiqi Jiedu prescription groups，control group， and prednisone group，with seven rats in each group， which were then gavaged with the corresponding drugs for one week for preparing the drug-containing serum. The effect of Yiqi Jiedu prescription-containing serum at different concentrations on the proliferation of mTEC and Treg cells were determined by cell counting kit-8 （CCK-8） assay. Besides， the effect of mTEC and Yiqi Jiedu prescription-containing serum on Treg cell proliferation were observed through co-culture. ResultThymocytes were cultured for a period of time. Their mean positive rate revealed by flow cytometry using mTEC characteristic marker Ulex europaeus agglutinin Ⅰ （UEAI） was 92.54%. Treg cells were sorted by magnetic beads. The purity of Treg cells after repeated magnetic bead sorting was as high as 92%. mTEC and Treg cells showed high positive expression rates，and their cell purity met the requirements of subsequent experiments. When the concentration of Yiqi Jiedu prescription-containing serum was 2.5%-15%，it exhibited an inhibitory effect against mTEC and Treg cells. When the concentration was equal to or greater than 20%，it promoted cell proliferation，which was further enhanced with the extension of action time. The results after 48 h of culture showed that compared with the control group，prednisone and low-dose Yiqi Jiedu prescription had no significant effect on the proliferation of these two kinds of cells，but the medium- and high-dose Yiqi Jiedu prescription remarkably reduced their proliferation inhibition rate （P<0.01）. After co-culture with mTEC， the control group was not significantly different from the prednisone group and the low-dose Yiqi Jiedu prescription-containing serum group in the proliferation of Treg cells，while the medium- and high-dose Yiqi Jiedu prescription-containing serum groups significantly lowered the proliferation inhibition rate （P<0.01）. ConclusionYiqi Jiedu prescription-containing serum affects the proliferation of mTEC and Treg cells in MG patients with thymus hyperplasia. Compared with the solely cultured Treg cells isolated from MG patients，the Treg cells co-cultured with mTEC exhibit enhanced proliferation in MG patients，suggesting that mTEC can regulate the proliferation of Treg cells. This effect becomes more obvious after the intervention with Yiqi Jiedu prescription-containing serum，indicating that intervention effect of Yiqi Jiedu prescription on Treg cells can be produced during its treatment of mTEC， which may be one of the mechanisms of Yiqi Jiedu prescription-containing serum in alleviating MG.

The apoptosis of mono-hepatocellular induced by the active ingredients of the Zanthoxyli Radix was investigated using laser Raman spectroscopy. Hepatoma cells (BEL-7404) were treated with 10 mg•L⁻¹ nitidine chloride and 3 g•L⁻¹ the extracts of Zanthoxyli Radix, respectively, then were divided into two parts, one for fluorescence staining, the other for determination of Raman spectroscopy. The acquired spectra were then processed by background elimination, smoothing, and normalization. Fluorescence staining results showed that the nucleuses from untreated group were uniformly stained, while those from the group treated for 48 hours were densely stained and broken. The spectra results revealed that the intensity of peaks associated with nucleic acid and protein decreased after the cells were incubated with the extracts of Zanthoxyli Radix for 12, 24, 36 and 48 hours. The intensity of peaks at 785,1 002,1 175,1 660 cm⁻¹ was decreased with the time of the cells were incubated by the extracts of Zanthoxyli Radix. The results indicated that the extracts of Zanthoxyli Radix could induce the apoptosis of hepatoma cells and reduce the amount of nucleic acid and protein in the cells. There is a certain relevance between the drug treatment time and the efficacy. The above results suggest that Raman spectra can provide abundant information about the changes in biological macromolecules within the cells after incubated by the extracts of Zanthoxyli Radix and serve as an effective method for the real time measurement of apoptosis.

Objective:To analyze the general situation and traditional Chinese medicine (TCM) syndrome characteristics in patients with coronavirus disease-2019（COVID-19） in Anhui province, and to provide a basis for clinical diagnosis and treatment. Method:The 144 cases of COVID-19 patients in Anhui province were collected from designated hospitals by means of multi-center cross-sectional epidemiological survey. The TCM syndrome information collection table of patients with novel coronavirus pneumonia was created to collect the general data, etiology and pathogenesis, pulse and signs, syndrome type and frequency statistics of the patients, and then summarize and analyze the main symptoms and syndrome distribution characteristics of patients with novel coronavirus pneumonia. Result:The mean age of 144 patients with COVID-19 infection was （43.54±15.91） years old, and the number of males was larger than the number of females (9/7). The age distribution and gender difference of the patients were statistically significant (P<0.05). 36.11%（52/144） of the patients had a history of travel/residence in Wuhan/Hubei province, and 63.89% (92/144) of the patients had a history of close contact with the confirmed patients. The main clinical manifestations included fever, cough, diarrhea, fatigue and poor tolerance. Light red tongue and red tongue were dominant in tongue quality, with mainly greasy coating, slippery pulse, rapid pulse and soft pulse were the main types of pulse. The main types of syndrome differentiation were the common type (76.38%, 110/144), most of which were demonstrated as dampness obstructing the lung and spleen (56.25%, 81/144). There was no significant difference in gender composition and age distribution between two groups. Conclusion:The pathogenesis of COVID-19 patients in Anhui province is closely related to the lung and spleen, and the dampness caused by pathogen of the epidemic virus is the main pathological factor of disease, which is in line with characteristics of dampness epidemic in TCM, also can see concurrently "heat, poison, stasis" for a characteristic.