Objective:To establish a three-dimensional culture and stress-loaded model of human periodontal ligament cells(hP-DLCs)in vitro and to determine the effect of static pressure on autophagy.Methods:Periodontal tissue was scraped from fresh ex-tracted tooth surfaces for culture and hPDLCs were obtained.Type Ⅰ rat tail collagen was extracted to prepare collagen-sodium algi-nate composite hydrogel,in which third-generation hPDLCs were embedded.After 72 h of culture,gene expression of autophagy-related proteins LC3,Atg-5 and Beclin-1 was detected by quantitative real-time polymerase chain reaction(qRT-PCR),and the expression of Atg-5,Beclin-1 and LC3 was detected by Western blotting.Results:Cells grew well in collagen-alginate hydrogels.After 15 min of compressive stress loading,the level of autophagy of hPDLCs increased significantly and then gradually decreased to the baseline level.Conclusion:Collagen-alginate hydrogel has good biocompatibility and safety,and can be used as an excellent scaffold for three-dimensional cell culture in vitro.Static compressive stress can accelevate autophagy,and this acceleration is rapid and short-lived.

This case report introduces Professor ZOU Wei 's experience of treating a patient with Wernicke encephalopathy using the "regulating spirit and promoting yang acupuncture method". The patient was diagnosed as spleen and stomach deficiency with internal liver wind. The treatment principle focused on regulating spirit and awakening the brain, strengthening the spleen, calming wind, and relaxing the tendons. Three groups of acupoints were selected: ①acupoints for awakening the brain by regulating spirit and unblocking meridians (Baihui [GV20], Qianshencong [EX-HN1] and bilateral Taiyang [EX-HN5], Fengchi [GB20]), etc.; ②acupoints for harmonizing the spleen, stomach, qi, and blood (bilateral Tianshu [ST25], Daheng [SP15], Taixi [KI3], etc.); ③acupoints for relaxing and softening the tendons (bilateral Waiguan [TE5], Hegu [LI4], Yanglingquan [GB34], Xuanzhong [GB39]).The needles were retained for 50 min per session, once daily, 7 days a week. After 16-week treatment, the patient was able to walk a few steps slowly with assistance, and other symptoms returned to normal. A two-month follow-up showed the patient's condition remained stable, walking distance further increased, and overall health significantly improved.
Humans ; Acupuncture Points ; Acupuncture Therapy ; Wernicke Encephalopathy/physiopathology*

Objective:To establish a three-dimensional culture and stress-loaded model of human periodontal ligament cells(hP-DLCs)in vitro and to determine the effect of static pressure on autophagy.Methods:Periodontal tissue was scraped from fresh ex-tracted tooth surfaces for culture and hPDLCs were obtained.Type Ⅰ rat tail collagen was extracted to prepare collagen-sodium algi-nate composite hydrogel,in which third-generation hPDLCs were embedded.After 72 h of culture,gene expression of autophagy-related proteins LC3,Atg-5 and Beclin-1 was detected by quantitative real-time polymerase chain reaction(qRT-PCR),and the expression of Atg-5,Beclin-1 and LC3 was detected by Western blotting.Results:Cells grew well in collagen-alginate hydrogels.After 15 min of compressive stress loading,the level of autophagy of hPDLCs increased significantly and then gradually decreased to the baseline level.Conclusion:Collagen-alginate hydrogel has good biocompatibility and safety,and can be used as an excellent scaffold for three-dimensional cell culture in vitro.Static compressive stress can accelevate autophagy,and this acceleration is rapid and short-lived.

Objective:To investigate the clinical value of targeted sequencing panel in the detection of genetic variation in neonates in neonatal intensive care unit (NICU).Methods:All neonates (≤28 d of age) admitted in the NICU (case group) and 200 full-term healthy neonates born with no obvious phenotypic abnormalities of Huzhou Maternity and Child Health Care Hospital were enrolled in this prospective study from November 2022 to January 2023. Based on a list of preventable and treatable rare diseases as well as newly screened diseases in China, a targeted sequencing panel suitable for Chinese newborns was designed to target the pathogenic genes and mutation sites associated with 601 genes and 542 diseases. Dried blood spot specimens were prepared and analyzed by the targeted sequencing panel. Pathogenic sites detected by the panel sequencing were verified using Sanger sequencing. The genetic testing results were analyzed according to the clinical features of the neonates. According to the number of primary clinical diagnosis index (including premature infants, neonatal hyperbilirubinemia, hemorrhagic diseases, neonatal infections, ventricular septal defect/patent ductus arteriosus, and others), these patients were divided into four groups with 1, 2, 3, and ≥4 diagnosis index, respectively. Chi-square test and linear correlation Chi-square test were used for statistical analysis. Results:There were 173 patients in the case group and 30.6% (53/173) of them carried pathogenic variants, including 52 positive for pathogenic genes and one with chromosome copy number variant. The positive rate of pathogenic genes was significantly higher in the case group than in the control group [30.1% (52/173) vs. 15.0% (30/200), χ 2=12.26, P<0.001]. Fourteen pathogenic genes were detected in the case group, including FLG, UGT1A1, G6PD, MYH7, AR, ABCC2, ACADS, CYP21A2, GJB2, MEFV, PAH, PKHD1, SCN4A, and HBA. In the case group, the detection rate of pathogenic variants in jaundiced neonates was higher than that in non-jaundiced neonates [35.2% (44/125) vs. 18.8% (9/48), χ 2=4.42, P=0.036]. However, there were no statistically significant differences in the detection rates of pathogenic variants between male and female infants, infants born to mothers of advanced maternal age or not, infants born to mothers with or without gestational diabetes mellitus, premature and term infants, or infants with or without hemorrhagic disorders, neonatal infections, or ventricular septal defects/patent ductus arteriosus in the case group (all P>0.05). The detection rate of pathogenic variants showed a linear increase in infants with 1, 2, 3, and ≥4 diagnosis index [21.1% (8/38), 25.4% (15/59), 38.2% (13/34), and 40.5% (17/42); linear correlation χ 2=4.84, P=0.028]. In the case group, seven genes with a high detection rate of genetic variation (including positive pathogenic genes and carriers) were UGT1A1 [had the highest detection rate, 24.9% (43/173)], GJB2, FLG, DUOX2, ABCA4, G6PD, and MUT. Seven loci with higher mutation frequency were c.211G>A(p.Gly71Arg), c.1091C>T(p.Pro364Leu), c.-41_-40dupTA, and c.686C>A(p.Pro229Gln) in the UGT1A1 gene, c.109G>A(p.Val37Ile) in the GJB2 gene, and c.12064A>T(p.Lys4022Ter) and c.3321del(p.Gly1109GlufsTer13) in the FLG gene. Conclusion:This panel sequencing can provide effective genetic testing for neonates in NICU, especially in children with complex clinical diagnosis.

Objective:To analyze the types and distribution of pathogenic variants for neonatal genetic diseases in Huzhou, Zhejiang Province.Methods:One thousand neonates (48 ~ 42 h after birth) born to Huzhou region were selected as the study subjects. Dry blood spot samples were collected from the newborns, and targeted capture high-throughput sequencing was carried out for pathogenic genes underlying 542 inherited diseases. Candidate variants were verified by Sanger sequencing.Results:Among the 1 000 newborns, the male to female ratio was 1.02 : 1.00. No pathogenic variants were detected in 253 cases, whilst 747 cases were found to carry at least one pathogenic variant, which yielded a carrier rate of 74.7%. The most frequently involved pathogenic gene was FLG, followed by GJB2, UGT1A1, USH2A and DUOX2. The variants were classified as homozygous, compound heterozygous, and hemizygous variants. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), 213 neonates were verified to have carried pathogenic and/or likely pathogenic variants, with a positive rate of 21.3%. The most commonly involved genes had included UGT1A1, FLG, GJB2, MEFV and G6PD. Conclusion:Newborn screening based on high-throughput sequencing technology can expand the scope of screening and improve the positive predictive value. Genetic counseling based on the results can improve the patients′ medical care and reduce neonatal mortality and childhood morbidity, while provide assistance to family members′ health management and reproductive decisions.

OBJECTIVE: To compare the clinical curative effect of muscle tension staged acupuncture and conventional acupuncture in the treatment of stroke hemiplegia. METHODS: Sixty-two patients with stroke hemiplegia were randomly divided into an observation group and a control group, 31 cases in each one. In the observation group, the muscle tension staged acupuncture was given, the six stages of Brunnstrom were classified as relaxation period and spasmodic period. The (consciousness-restoring resuscitation) combined with the hand and foot meridian acupuncture were applied at Shuigou (GV 26), Jianyu (LI 15), Quchi (LI 11), Shousanli (LI 10), Hegu (LI 4), Liangqiu (ST 34), Zusanli (ST 36), Shangjuxu (ST 37), Jiexi (ST 41) during relaxation period; mainly by hand and foot meridian and meridian, the acupoints were Jianliao (TE 14), Tianjing (TE 10), Waiguan (TE 5), Yangchi (TE 4), Houxi (SI 3), Huantiao (GB 30), Yanglingquan (GB 34), Chengshan (BL 57), Xuanzhong (GB 39), Shenmai (BL 62), Qiuxu (GB 40) during spasmodic period. In the control group, referring to 's , mainly by hand meridian, the governor vessel and foot meridian, phasing was not considered in the acupuncture treatment plan. Both groups were treated one time a day for 4 weeks. The neurological deficit scores were observed before and after treatment of the two groups and the efficacy was evaluated. RESULTS: There was one case dropped in each group. After treatment, the neurological deficit scores of the two groups was lower than those before treatment (both <0.05), and the observation group was lower than the control group (<0.05). The cured and markedly effective rate was 66.7% (20/30) in the observation group, which was higher than 36.7% (11/30) in the control group, the difference between the two groups was statistically significant (<0.05). CONCLUSION The muscle tension staged acupuncture is better than the conventional acupuncture for the treatment of stroke hemiplegia.
Acupuncture Points ; Acupuncture Therapy ; Hemiplegia ; therapy ; Humans ; Muscle Tonus ; Stroke ; Treatment Outcome

OBJECTIVE: To observe the effectiveness of acupuncture on claustrophobia, and to explore the effects of acupoint specificity on claustrophobia. METHODS: This was an evaluator-blinded randomized controlled trial. One hundred and sixty patients who presented with claustrophobia during magnetic resonance imaging examination were randomized into an acupoint group, a non-acupoint group, a sham-acupoint group and a blank group, 40 cases in each one. The patients in the acupoint group were treated with acupuncture at Zhaohai (KI 6), Taichong (LR 3), Lingdao (HT 4), Neiguan (PC 6), Shenmen (HT 7), Danzhong (CV 17), Baihui (GV 20) and Fengchi (GB 20). The patients in the non-acupoint group were treated with acupuncture at points 0.5 next to the acupoints above. The patients in the sham-acupoint group were treated with acupuncture at acupoints not closely correlated to claustrophobia in corresponding segment. All the acupuncture was given once. No treatment was used in the blank group. The state anxiety questionnaire (S-AI) was observed in all the patients at the end of MRI examination before and after treatment. The clinical therapeutic effects were compared among four groups. RESULTS: Compared before treatment, the S-AI score was reduced in the acupoint group, non-acupoint group and sham-acupoint group after treatment (<0.05, <0.01); the differences of S-AI score before and after treatment in the blank group were not statistically significant (>0.05). After treatment, the S-AI scores in the acupoint group, non-acupoint group and sham-acupoint group was lower than that in the blank group (<0.05, <0.01), and the differences of S-AI score were higher than that in the blank group (<0.01). The S-AI score in the acupoint group was lower than that in the non-acupoint group and sham-acupoint group (<0.05), and the difference of S-AI score was higher than those in the non-acupoint group and sham-acupoint group (<0.05). The difference of S-AI score in the non-acupoint group was higher than that in sham-acupoint group (<0.05). The total effective rate was 92.5% (37/40) in the acupoint group, which was significantly superior to 25.0% (10/40) in the non- acupoint group, 17.5% (7/40) in the sham-acupoint group and 5.0% (2/40) in the blank group (<0.05, <0.01). CONCLUSION Acupuncture showes superior effect on claustrophobia, and its tranquilizing effect may be related with acupoint specificity.
Acupuncture Points ; Anxiety ; Humans ; Phobic Disorders ; therapy ; Surveys and Questionnaires

Objective To observe the clinical curative effect of Zoladex combined with Sanjie analgesic capsule in treatment of endometriosis.Methods 80 patients with endometriosis treated in our hospital from January 2016 to December 2016 were randomly divided into two groups,with 40 cases in each group.The study group was given Zoladex needle and Sanjiezhentong capsule treatment;The control group were given Sanjie Zhentong capsule.Prostaglandin F2a(PCF2a), CA125, visual analogue scale(VAS), clinical efficacy and adverse reactions were compared between the two groups.Results After treatment, the PCF2a and CA125 in the study group were significantly higher than those in the control group, the difference was statistically significant(t=4.167, t=3.533, P<0.05).There was no significant difference in VAS score between the two groups(t=0.961, P>0.05).The total effective rate of the study group was 90%, and the control group was 75%, the difference was statistically significant(x2=5.741, P<0.05)., there were 4 cases of adverse reactions(10%)in the study group,and 10 cases in the control group(25%), the difference was statistically significant(x2=7.792, P<0.05).Conclusion Zoladex needle combined with Sanjiezhentong capsule, can effectively relieve the pain caused by endometriosis, reduce adverse reaction, improve the clinical efficacy, suitable for use in clinical research.

ObjectiveTo investigate the clinical efficacy of acupuncture in treating Tourette's syndrome.MethodEighty patients were allocated, using a random number table, to a treatment group of 40 cases and a control group of 40 cases. The treatment group received acupuncture and the control group, Western drug treatment.ResultThe YGTSS score decreased significantly in the two groups of patients after treatment(P<0.01). After treatment, the range of decrease in the YGTSS tic score was larger in the treatment group than in the control group; there was a statistically significant difference (P<0.05). The total efficacy rate was higher in the treatment group than in the control group (P<0.05). The incidences of adverse reactions were significantly lower in the treatment group than in the control group.ConclusionThe effect and safety of treatment for Tourette's syndrome are higher in the treatment group than in the control group.

Toutianliang manipulation is the most representative compound needling technique of traditional reinforcing and reducing and has the remarkable efficacy on heat syndrome with muscle and bone involved for example. Professor ZHANG Jin is one of the famous acupuncture master in China and has contributed his lifelong to the research of acupuncture manipulation techniques. He has summarized 24 single-type manipulations and has given the comprehensive explanation of the manipulations for meridian conduction, reinforcing or reducing techniques for the excess or deficiency. In the paper, Toutianliang manipulation was introduced briefly and the key operation steps had been discussed.
Acupuncture Therapy ; history ; methods ; China ; History, 20th Century ; History, 21st Century ; Humans